Objective To observe the morphological and functional changes of different lung cells in hyperoxia environment.Methods Type Ⅱ alveolar epithelial cells (AECⅡ) and lung fibroblasts (LFs) of fetal rats with 18 days old were isolated and culturedin vitro, and divided into air group (placed in an atmospheric incubator, and culturing with oxygen volume fraction of 0.21) and hyperoxia group (placed in a high oxygen culture chamber, and culturing with oxygen volume fraction of 0.90). Morphological changes of two kinds of cells were observed under microscope. Cell migration was observed by scratch test. The levels of reactive oxygen species (ROS) and apoptosis in cells were detected by flow cytometry.Results After 8 hours of hyperoxia, the volume of AECⅡincreased and the cells were loosely arranged; the clearance of LFs cells was increased and arranged in disorder. Scratch test showed that, compared with air group, the immigration rate of AECⅡ was inhibited at 6 hours hyperoxia [migration rate: (38.67±1.15)％ vs. (58.67±2.31)％,P < 0.01], the immigration rate of LFs was promoted at 12 hours after hyperoxia [migration rate:(55.37±1.50)％ vs. (46.90±1.20)％,P < 0.01]. With the increase of hyperoxia time, intracellular ROS contents of two cells were gradually increased, which were significantly higher than those of the air group (fluorescence intensity:130.67±4.04 vs. 54.67±2.51, 85.00±2.00 vs. 60.33±1.52, bothP < 0.01). Both two kinds of cells showed apoptosis after exposure to high oxygen, the apoptosis rate of AECⅡ at 2 hour exposure were significantly higher than that of air group [(1.93±0.28)％ vs. (1.07±0.11)％,P < 0.05], the apoptosis rate of LFs at 6 hour exposure was significantly higher than that of air group [(1.66±0.09)％ vs. (1.46±0.09)％,P < 0.05].Conclusion High concentration of oxygen can cause poor growth of lung cells, reduce AEC Ⅱ migration level and increase LFs migration, and the production of intracellular ROS eventually leads to apoptosis of lung cells.

OBJECTIVEYoung C57BL/6 (B6) mice were treated with a specific tolerogen-dual analogue (Lys262-Ala207) intranasally to observe its effect on the invasion process of mice model and the clinical symptoms, to assess its clinical effects, and to explore the underlying mechanisms and feasibility of nasal mucosal tolerance explored.

METHODSPassively transferred myasthenia gravis (PTMG) was induced by mAb35 on B6 young female mice. Sixty mice were divided equally into three groups: tolerance group, model group and control group. Lys262-Ala207 was given intranasally (250 microg/mouse) to tolerance group with mAb35 for 10 successive days before immunization. Model group received PBS 50 microl only. The body weight and clinical scores were evaluated. The serum levels of AChRAb and the main cytokines (IL-4, IFN-gamma, TGF-beta1) were detected with ELISA.

RESULTSThe model group had typical myasthenia symptoms. B6 mice of tolerance group had less severe symptoms compared with control groups. The clinical symptoms of tolerance group were relieved. The level of AChRAb in tolerance group [(16.01 +/- 1.09) mg/L] was significantly lower than that of model group [(28.12 +/- 1.28) mg/L] (t = 44.37, P < 0.01). IL-4 and IFN-gamma levels in tolerance group [(141.02 +/- 3.11) ng/L, (187.99 +/- 4.67) ng/L] were significantly lower than those of model group [(193.37 +/- 3.95) ng/L, (320.46 +/- 2.14) ng/L] (t = 37.20, 51.69, P < 0.01). The level of TGF-beta1 in tolerance group [(437.19 +/- 1.93) ng/L] was higher than that of model group [(175.63 +/- 3.12) ng/L] (t = 36.07, P < 0.01). But there were still significant change as compared to those in control group (t = 26.65, 31.05, 49.02, P < 0.01).

CONCLUSIONSNasal administration of Lys262-Ala207 ameliorated muscular weakness in PTMG young mice. The therapeutic effect is possibly correlated with the function of immune system.

Animals ; Female ; Immune Tolerance ; Immunity, Mucosal ; Interleukin-4 ; blood ; Mice ; Mice, Inbred C57BL ; Myasthenia Gravis ; blood ; immunology ; Nasal Mucosa ; immunology ; Transforming Growth Factor beta1 ; blood

The nitrite-reducing activity of aerobic denitrifying bacterial strain N6-1 was studied. It showed that the nitrite-reducing activity reached the highest at 30℃, 120 r/min, pH 8.5 and C/N ratio 12, using CH3COONa and NaNO2 as the sole carbon source and nitrogen source, respectively. When the initial NaNO2 concentration was 2 g/L, NO2--N was reduced completely after 20 hours cultivation with the reducing rate of 20.3 mg/L?h. There would be no effect on its nitrite-reducing activity in the present of 1.5% NaCl or 1% peptone. The cell concentration could reach 1.2?1011 CFU/mL after 24 hours cultivation in 10 L fermentor.

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.

To clone the 2-C-methyl-D-erythritol 4-phosphate cytidylyltransferase (TwMCT) full length cDNA from Tripterygium wilfordii, the specific primers were designed according to the transcriptome data and the LCPCR were carried out. After a series of bioinformatics analysis on the TwMCT, the MeJA induced expression content were investigated by real-time fluorescence quantification polymerase chain reaction (RT-qPCR). The result showed that the full of TwMCTcDNA was 1 318 bp nucleotides encoding 311 amino acids. The molecular weight of the deduced TwMCT protein was about 34.14 kDa and the theoretical isoelectric point was 8.65. Result of the RT-qPCR analysis indicated that the content of TwMCT mRNA expression in T. wilfordii suspension cell was rising after treating with MeJA and reached the maximum in 24 h. Cloning and analyzing TwMCT gene from T. wilfordii provided gene element for studying the function and expression regulation of secondary metabolites.
Amino Acid Sequence ; Cloning, Molecular ; Erythritol ; analogs & derivatives ; metabolism ; Gene Expression Regulation, Plant ; Molecular Sequence Data ; Nucleotidyltransferases ; chemistry ; genetics ; metabolism ; Phylogeny ; Plant Proteins ; chemistry ; genetics ; metabolism ; Protein Structure, Secondary ; Sequence Alignment ; Sugar Phosphates ; metabolism ; Tripterygium ; chemistry ; enzymology ; genetics

Objective To investigate the prevalence of diabetes and impaired glucose regulation in high risks community residents in Songjiang District of Shanghai, and to explore the risk factors affecting their blood glucose metabolism, providing effective suggestions for improving community diabetes prevention and treatment. Methods Questionnaire, physical examination and laboratory testing were used to collect information on the basic characteristics and blood glucose levels of 21 035 residents in Songjiang District who were assessed to be at high risk of diabetes. Results A total of 3 008 people with impaired regulation and 2 241 patients with diabetes were detected.The detection rates were 14.3% and 10.6%, respectively.It was found that as the residents′ age was higher, their education level was lower, high-risk factors for them were numerous, and their detection rates of diabetes and impaired sugar regulation were higher.It was found that age, gender, history of impaired glucose regulation, relatives with type 2 diabetes, hypertension, dyslipidemia and overweight/obesity were all factors influencing blood glucose metabolism (P < 0.05). Conclusion Blood glucose screening in high-risk population of diabetes is helpful for the early detection of diabetic patients and those with impaired glucose regulation.It is necessary to strengthen the attention of population at risk of diabetes and take appropriate interventions.

A full-length cDNA of GGPPS gene from Tripterygium wilfordii suspension cells was obtained by use of RACE strategy (GeneBank: KM978333), and then analyzed by bioinformatics approaches. TwGGPPS cDNA has 1857 nucleotides and an open reading frame (ORF) encoding a protein of 514 amino acid residues. The deduced protein has isoelectric point (pI) of 7.85, a calculated molecular weight about 57.13 kD, 5 conserved domains and 2 functional domains. PSORT Prediction showed it was located at plasma membrane. Phylogenetic analysis demonstrated that TwGGPPS1 was similar to GGPPS from other species of plants. For the first time the cloning of geranylgeranyl diphosphate synthase gene from T. wilfordii was reported, it lays the foundation for further research of diterpenoids biosynthetic pathway.
Amino Acid Sequence ; Cloning, Molecular ; Farnesyltranstransferase ; chemistry ; genetics ; metabolism ; Molecular Sequence Data ; Phylogeny ; Plant Proteins ; chemistry ; genetics ; metabolism ; Sequence Alignment ; Sequence Homology, Amino Acid ; Tripterygium ; chemistry ; enzymology ; genetics

Weight loss is frequently observed after acute exposure to high altitude. However, the magnitude and rate of weight loss during acute exposure to high altitude has not been clarified in a controlled prospective study. The present study was performed to evaluate weight loss at high altitude. A group of 120 male subjects [aged (32±6) years] who worked on the construction of the Golmud-Lhasa Railway at Kunlun Mountain (altitude of 4 678 m) served as volunteer subjects for this study. Eighty-five workers normally resided at sea level (sea level group) and 35 normally resided at an altitude of 2 200 m (moderate altitude group). Body weight, body mass index (BMI), and waist circumference were measured in all subjects after a 7-day stay at Golmud (altitude of 2 800 m, baseline measurements). Measurements were repeated after 33-day working on Kunlun Mountain. In order to examine the daily rate of weight loss at high altitude, body weight was measured in 20 subjects from the sea level group (sea level subset group) each morning before breakfast for 33 d at Kunlun Mountain. According to guidelines established by the Lake Louise acute mountain sickness (AMS) consensus report, each subject completed an AMS self-report questionnaire two days after arriving at Kunlun Mountain. After 33-day stay at an altitude of 4 678 m, the average weight loss for the sea level group was 10.4% (range 6.5% to 29%), while the average for the moderate altitude group was 2.2% (-2% to 9.1%). The degree of weight loss (Δ weight loss) after a 33-day stay at an altitude of 4 678 m was significantly correlated with baseline body weight in the sea level group (r=0.677, P<0.01), while the correlation was absent in the moderate altitude group (r=0.296, P>0.05). In the sea level subset group, a significant weight loss was observed within 20 d, but the weight remained stable thereafter. AMS-score at high altitude was significantly higher in the sea level group (4.69±2.48) than that in the moderate altitude group (2.97±1.38), and was significantly correlated with baseline body weight. These results indicate that (1) the person with higher body weight during stay at high altitude loses more weight, and this is more pronounced in sea level natives when compared with that in moderate altitude natives; (2) heavier individuals are more likely to develop AMS than leaner individuals during exposure to high-altitude hypoxia.
Adult ; Altitude ; Altitude Sickness ; physiopathology ; Body Mass Index ; Body Weight ; China ; Humans ; Hypoxia ; physiopathology ; Male ; Weight Loss ; physiology

OBJECTIVETo evaluate the effect of epilepsy on sleep in children.

METHODSWhole night polysomnography was performed in 48 epileptic children and 12 healthy controls. The 48 epileptic children were divided into focal seizure and generalized seizure groups and into waking seizure and sleeping seizure groups according to the time of occurrence of the seizures. Various parameters of sleep structure were analyzed.

RESULTSThe sleep efficiency of epileptic children was significantly lower than that of the healthy controls (85.4 +/- 8.6% vs 90.9 +/- 5.8%; P < 0.05). The total recording time (TRT) of sleep was significantly longer and the sleep efficiency was significantly lower in the focal seizure group compared to the control group (P < 0.05). The percentage of stage 1 non-rapid-eye-movement sleep (S1 sleep) increased and the percentage of rapid-eye-movement (REM) sleep decreased in the generalized seizure group compared to the control group (P < 0.05). The percentage of S1 sleep increased and both the percentage of REM sleep and the sleep efficiency decreased in the sleeping seizure group as compared with the control group (P < 0.05). There were no significant differences in the parameters of sleep structure between the waking seizure and the control group. Among the sleeping seizure group, the children with generalized seizure showed significantly lower REM sleep percentage and sleep efficiency, and those with focal seizure had significantly longer TRT and higher S1 sleep percentage as compared with the controls.

CONCLUSIONSEpilepsy affects sleep structure of patients, and different types of seizure have different influences on sleep structure. Children with generalized seizure have prolonged light sleep and shortened REM sleep. When generalized seizures occur during waking, the increase of light sleep is more pronounced. While generalized seizures occur during sleeping, REM sleep reduction is more prominent. Children with focal seizures have decreased sleep efficiency. When focal seizures occur during waking, the sleep structure of patients is normal. However, when seizures occur during sleeping light sleep increases and sleep efficiency decreases.

Adolescent ; Child ; Child, Preschool ; Epilepsy ; physiopathology ; Female ; Humans ; Infant ; Male ; Polysomnography ; Sleep Stages ; physiology

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