BACKGROUND: This study aimed to investigate the prevalence rate of critical illness-related corticosteroid insufficiency (CIRCI) and the effect of low-dose glucocorticoid on prognosis of CIRCI in patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD). METHODS: Since January 2010 to December 2012, 385 patients, who met the criteria of AECOPD, were enrolled in the Intensive Care Unit (ICU) of the First People's Hospital and Municipal Central Hospital of Xiangtan City. The AECOPD patients complicated with CIRCI screened by an adrenalcorticotrophic hormone test within 12 hours after admission to ICU were divided into a treatment group (n=32) and a control group (n=31) for a prospective, randomized and controlled clinical trial. Hydrocortisone (150 mg/d) or normal saline was injected intravenously for 7 days. The patients were followed up for 28 days after injection. The endpoint included 28-day survival time, non-shock time, ICU stay and the period of non-mechanical ventilation. The markers of inflammation C-reactive protein, tumor necrosis factor-α, interleukin 6 and procalcitonin were measured at baseline and 7 days after treatment. The variables were analyzed by Student's t test, the non-parametric statistical test, the Chi-square test or the Kaplan-Meier method with SPSS18.0 statistic software. A P value <0.05 was considered statistically significant. RESULTS: Totally 63 patients were diagnosed with CIRCI by an adrenalcorticotrophic hormone test and the prevalence rate was 16.4％. The shock rate of the AECOPD patients complicated with CIRCI was higher than that of the AECOPD patients without CIRCI (23.8％ vs. 8.7％, P<0.01). Kaplan-Meier analysis revealed that the 28-day survival time of the treatment group was obviously longer than that of the control group (P<0.05). Compared with the control group, shock-free days within 28 days was longer in the treatment group (18.2±9.5 vs. 25.8±4.1, P<0.05). Treatment with low-dose glucocorticoid obviously decreased the markers of infection and inflammation (P<0.01), such as C-reactive protein (13.2±5.5 mg/L vs. 8.3±3.1 mg/L for the control group; 13.5±5.9 mg/L vs. 5.1±2.3 mg/L for the treatment group), tumor necrosis factor-α (26.1±16.2 g/L vs. 17.5±11.7 g/L for the control group; 25.0±14.8 g/L vs. 10.4±7.8 g/L for the treatment group) and procalcitonin (3.88 g/L vs. 2.03 g/L for the control group; 3.77 g/L vs. 1.26 g/L for the treatment group). Furthermore, the markers in the treatment group decreased more obviously than those in the control group (P<0.01). CONCLUSION: The prevalence rate of CIRCI was higher in the patients with AECOPD in the department of critical medicine, and low-dose glucocorticoid treatment for one week reduced the 28-day mortality, shock time and markers of infection and inflammation.

OBJECTIVETo compare the difference in the clinical efficacy on cervical spondylosis of vertebral artery type (CSA) treated with thermosensitive moxibustion at different dosages.

METHODSSixty cases of CSA were randomized into a saturated moxa dosage group and a regular moxa dosage group, 30 cases in each one. The thermosensitive moxibustion was adopted in the two groups. The mild suspended moxibustion was applied at two acupoints with the strongest thermosensitization. In the saturated moxa dosage group, the moxibustion time was determined by the disappearance of thermosensitization. In the regular moxa dosage group, 15 min was required on each acupoint. The treatment was given twice a day for first 4 days in the two groups. Since the 5th day, the treatment was given once a day, continuously for 10 times, and totally 14 days were required. The score of symptoms and function and clinical efficacy were compared between the two groups before and after treatment as well as 6-month follow-up after treatment.

RESULTSThe curative and effective rate was 56.7% (17/30) after treatment and 60.0% (18/30) in 6-month follow-up after treatment in the saturated moxa dosage group, which were superior to 26.7% (8/30) and 30.0% (9/30) in the regular moxa dosage group respectively (P < 0.01, P < 0.05). The scores of clinical symptoms and function after treatment and in follow-up were improved apparently as compared with those before treatment in both groups (all P < 0.01). The scores of clinical symptoms and function after treatment and in follow-up in the saturated moxa dosage group were increased much more apparently than those in the regular moxa dosage group (after treatment: 22.32 +/- 4.64 vs 17.43 +/- 3.21; in follow-up: 23.01 +/- 4.76 vs 18.32 +/- 2.13, both P < 0.01).

CONCLUSIONThe thermosensitization moxibustion of saturated dosage achieves the superior short-term and long-term efficacies in the treatment of CSA as compared with the regular moxibustion dosage.

Acupuncture Points ; Adult ; Female ; Humans ; Male ; Middle Aged ; Moxibustion ; instrumentation ; Spondylosis ; physiopathology ; therapy ; Vertebral Artery ; physiopathology

OBJECTIVETo observe the effect of Jiannao Yizhi Decoction (JNYZD) on learning and memory in rats with similar Alzheimer's disease (AD), and to investigate its possible mechanism.

METHODSThe composite AD rat model was established by injecting aggregated Abeta25-35 into the lateral cerebral ventricle of senile rats, and all the modeled rats were divided into 5 groups, the model group, the Donepezil group, the high-, middle-, and low-dose JNYZD group. All rats, except those in the model group, were treated respectively with Donepezil and JNYZD at the daily dose of 0.525 mg/kg, 42.4 g/kg, 21.2 g/kg, 10.6 g/kg for 21 days. The ability of learning and memory of rats in different groups was tested using Morris water maze, and the activity of acetylcholine esterase (AchE) and butyrocholin esterase (BehE) in serum were determined, too.

RESULTSThe escape latent period was shorter in all medicated group than in the model group (P<0.01 or P<0.05), and it was insignificantly different among all medicated groups (P>0.05). A decreasing trend of AchE and BchE activity presented in the high- and middle-dose JNYZD groups, but insignificant difference was shown as compared these indexes respectively with those in the Donepezil group. Furthermore, the improvement of learning and memory in similar AD rats was insignificantly different between the Donepezil group and the JNYZD groups (P>0.05).

CONCLUSIONJNYZD can improve the learning and memory ability of similar AD rats by influencing the activity of cholinesterase.

Acetylcholinesterase ; blood ; Alzheimer Disease ; blood ; physiopathology ; Animals ; Butyrylcholinesterase ; blood ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; pharmacology ; Escape Reaction ; drug effects ; Learning ; drug effects ; Male ; Maze Learning ; drug effects ; Memory ; drug effects ; Rats ; Rats, Wistar

To analyze components of Citrus reticulata and salt-processed C. reticulata by ultra-performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF/MS), and compared the changes in components before and after being processed with salt. Principal component analysis (PCA) and partial least squares discriminant analysis (OPLS-DA) were adopted to analyze the difference in fingerprint between crude and processed C. reticulata, showing increased content of eriocitrin, limonin, nomilin and obacunone increase in salt-processed C. reticulata. Potential chemical markers were identified as limonin, obacunone and nomilin, which could be used for distinguishing index components of crude and processed C. reticulata.
Benzoxepins ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Citrus ; chemistry ; Discriminant Analysis ; Limonins ; chemistry ; Mass Spectrometry ; methods ; Principal Component Analysis ; methods ; Salts ; chemistry

Objective To construct eukaryotic enhanced green fluorescent protein (EGFP) expressing recombinant plasmid, pEGFP-C1-hnRNP A1, which contains coding sequence of human hnRNP A1 (heterogeneous nuclear ribonucleo-protein A1), and to perform cellular localization analysis of EGFP tagged hnRNP A1 under stress. Methods Total RNA was isolated from HeLa cell used for synthesis of first-strand cDNAs using reverse primers that are specific for the 3′-un-translated region of hnRNP A1. hnRNP A1 gene fragments were then amplified by touch-down PCR from those cDNAs and inserted into pEGFP-C1 fluorescent bearing vector through EcoRⅠ/BamHⅡdouble enzyme digestion and T4 DNA Ligase connection. The recombinant pEGFP-C1-hnRNP A1 plasmid was transfected into HeLa cells and green fluorescent tagged fusion proteins was examined by Western blot and confocal fluorescence microscopy. Co-localization of EGFP-hnRNP A 1 with poly (A)+mRNA (the marker of the stress granules), or DCP1a (the marker of processome) were detected by RNA fluores-cence in situ hybridization and immunofluorescence. Results The pEGFP-C1-hnRNP A1 was sequenced and digested cor-rectly by restriction single/double enzyme. The green fluorescent fusion protein was also detected in transfected HeLa cell by Western blot and confocal fluorescence microscopy. EGFP-hnRNP A1 co-localizes with poly(A)+mRNA, but not DCP1a. Conclusion Recombinant eukaryotic plasmid of pEGFP-C1-hnRNP A1 was constructed successfully and expressed effec-tively. EGFP tagged hnRNP A1 takes part in forming stress granules.

OBJECTIVETo investigate the influence of pleiotrophin (PTN) on the growth of rat hepatocytes.

METHODSPrimary rat hepatocytes were isolated from male Sprague-Dawley rats and divided into three groups: group A (negative control), cultivated in normal culture medium; group B (positive control), cultivated with culture medium supplemented with supernatant from the embryonic fibroblast 3T3 cell line; group C (experimental), cultivated with culture medium supplemented with human recombinant (hr) PTN (100 ng/ml). The hepatocytes' growth rate and level of secreted albumin (ALB) were evaluated by microscopy and biochemical assay, respectively. Significance of between-group differences were assessed by one-way ANOVA, and pairwise comparisons were performed by the least significant difference test.

RESULTSThe growth rates of hepatocytes in groups A, B and C were 2.800+/-0.084%, 4.300+/-0.132% and 3.800+/-0.053%, respectively. The growth rate of group B was significantly higher than the other two groups (F = 333.735, P less than 0.05). For all groups, the highest levels of secreted ALB were detected between the second and sixth day of culture, with g/L concentrations at day 2, 4 and 6 of: group A, 0.550+/-0.010, 0.900+/-0.030 and 0.300+/-0.040; group B, 0.900+/-0.030, 1.300+/-0.020 and 1.400+/-0.030; group C, 0.900+/-0.010, 1.160+/-0.010 and 0.700+/-0.050. The secreted ALB of group B was significantly higher than that of the other two groups (F = 651.355, 338.831 and 863.205, P less than 0.05 ).

CONCLUSIONPTN can benefit in vitro culturing of rat hepatocytes by stimulating growth and enhancing their ability to secrete albumin.

Albumins ; secretion ; Animals ; Carrier Proteins ; pharmacology ; Cells, Cultured ; Cytokines ; pharmacology ; Hepatocytes ; cytology ; drug effects ; secretion ; Male ; Rats ; Rats, Sprague-Dawley

BACKGROUND: Increasing evidence has indicated that low-concentration hydrogen or hydrogen rich water or hydrogen saturated saline exerts a protective effect on various diseases, such as myocardial ischemia/reperfusion injury. OBJECTIVE: To explore the protective effect of hydrogen rich water on myocardial ischemia/reperfusion injury. METHODS: Forty-eight Wistar rats were equally randomized into control and hydrogen-rich groups, and then subdivided into ischemic preconditioning, ischemia, and ischemia/reperfusion groups (n=8 rats in each subgroup). The myocardial ischemia/reperfusion model was established in the heart of each rat by the following procedures: reverse perfusion for 10 minutes, room temperature for 20 minutes, and reperfusion for 20 minutes. The control rats was perfused with pre-oxygenated (95% O2plus 5% CO2) 37 ℃ K-R solution and the hydrogen-rich group was perfused with pre-oxygen-equilibrated (95% O2plus 5% CO2) 37 ℃ K-R solution plus hydrogen-rich water (0.6 mmol/L, pH=7.3). Subsequently, the heart was removed, the pathological changes of the myocardial tissues were observe by hematoxylin-eosin staining, the activities of lactic dehydrogenase and creatine kinase in the myocardial tissues were determined, and the levels of tumor necrosis factor-α and interleukin-1β were detected by ELISA. RESULTS AND CONCLUSION: In the control group, the activity of lactic dehydrogenase at the ischemic and ischemia/reperfusion stages was significantly higher than that at the ischemic preconditioning stage (P < 0.05), and the activity of creatine kinase at the ischemia/reperfusion stage was significantly higher than that at the ischemic preconditioning and ischemic stages (P < 0.05). In the hydrogen-rich group, there was no significant difference in the activities of lactic dehydrodenase and creatine kinase at each stage, but the activities of at the ischemia/reperfusion stage was significantly lower than those in the control group (P < 0.05). In the two groups, the order of the levels of tumor necrosis factor-α and interleukin-1β was as follows: the ischemia/reperfusion stage > ischemic stage > ischemic preconditioning stage (P < 0.05). The levels of above factors in the hydrogen-rich group were significantly lower than those in the control group (P < 0.05). Our findings imply that hydrogen rich water has protective effect on myocardial ischemia/reperfusion injury of the rat hearts in vitro,which may be by reducing the expression of tumor necrosis factor-α and interleukin-1β, and further alleviating the inflammatory response.

Chronic myelogenous leukemia (CML) is a clonal myeloproliferative disease of transformed hematopoietic progenitor cells. The expressions of JunB and CDH13 (cadherin-13) gene as tumor suppressor gene were inactivated by promoter methylation in CML patients. This study was purposed to investigate the methylation difference of JunB and CDH13 gene promoter and the expression levels of JunB and CDH13 gene in CD34(+)CD38(-) cells in CML patients vs normal individuals. CD34(+)CD38(-) cells from 8 cases of CML and 5 normal individuals were selected by flow cytometry. The methylation status of JunB and CDH13 genes were detected by MS-PCR in selected CD34(+)CD38(-) cells. The expression levels of JunB and CDH13 gene was detected with real time polymerase chain reaction (RT-PCR). The results showed that no methylation of JunB and CDH13 gene was detected in CD34(+)CD38(-) cells of 5 normal individuals. Methylations of JunB and CDH13 promoter were found in 87.5% (7/8) and 50% (4/8) CML CD34(+)CD38(-) cells, percentages of which were significantly higher than those in normal individuals. The difference was statistically significant (p < 0.05). The relative expression levels of JunB and CDH13 mRNA in CD34(+)CD38(-) cells of CML patients were significantly lower than those in normal individuals (2(-DeltaDeltaCT) were 1/5.21 and 1/10.63 respectively). It is concluded that the high methylation of JunB and CDH13 gene promoter occurs in CD34(+)CD38(-) cells of CML patients, their mRNA expression level is significantly lower, thus the methylation of JunB and CDH13 gene promoter probably plays a role in the pathogenesis of CML and may have clinical significance in predicting prognosis of CML.
ADP-ribosyl Cyclase 1 ; immunology ; Adult ; Aged ; Antigens, CD34 ; immunology ; Cadherins ; genetics ; DNA Methylation ; Female ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; Male ; Middle Aged ; Promoter Regions, Genetic ; Proto-Oncogene Proteins c-jun ; genetics

OBJECTIVE: To investigate the correlations of four genetic single nucleotide polymorphisms (SNPs) of brain-derived neurotrophic factor (BDNF) with posttraumatic stress disorder (PTSD). METHODS: A total of 300 patients with sporadic PTSD and 150 healthy subjects (the control group) were selected according to the diagnostic criteria of PTSD (DSM-IV), and the genotypes of the BDNF SNPs G-712A, C270T, rs6265, and rs7103411 were detected by polymerase chain reaction and direct DNA sequencing to determine intergroup differences in the genotypes and allele frequencies; the p values were corrected with the permutation test. RESULTS: The genotypes and allele frequencies of the SNPs G-712A, rs6265, and rs7103411 of BDNF showed no significant intergroup differences (p>0.05). However, the genotype and allele frequencies of C270T showed significant differences between the PTSD group and the control group (p<0.05). CONCLUSION: The SNP C270T of BDNF may be associated with PTSD. Individuals carrying the polymorphic T allele of C270T may be more likely to suffer from PTSD.
Alleles ; Brain-Derived Neurotrophic Factor* ; Gene Frequency ; Genotype ; Healthy Volunteers ; Humans ; Polymerase Chain Reaction ; Polymorphism, Single Nucleotide* ; Sequence Analysis, DNA ; Stress Disorders, Post-Traumatic*

Anemarrhena asphodeloides processed by salt and raw product was compared including both chemical composition and laxative function in order to find the possible active substance to cure constipation. Processed and raw Anemarrhenae laxative effect on experimental constipation models was observed as well as chemical composition using UPLC-MS technology and the total sugar content was determined by phenol sulfuric acid method. Processed Anemarrhenae water extract improved excrement more than raw which has significant difference compared with the blank group (P < 0.05). On the other hand, the total ion flow spectrum showed no significant difference in most substance, but the total sugar content was significantly higher than raw product. Anemarrhenae ancient be recognized benefitting for draining body water in traditional Chinese medicine which has been lost in modern books because it is manifested as excellent laxative effect not diuretic effect. Saccharides carbohydrate may have closely relationship with this magically effect.
Anemarrhena ; chemistry ; Animals ; Chemistry, Pharmaceutical ; Constipation ; drug therapy ; physiopathology ; Defecation ; drug effects ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Humans ; Laxatives ; administration & dosage ; chemistry ; Male ; Rats, Wistar ; Rhizome ; chemistry