Objective:To establish the reference for serum metabolomics profiles among healthy Han adults in China, and explore the variation on metabolomics profiles by geographic regions, sex, and age.Methods:Cross-sectional data and serum samples were obtained from the China National Health Survey. A total of 1 039 male and 1 032 female healthy adults(≥30 years) were included in this study. Serum metabolomics analyses were conducted with ultra-performance liquid chromatography-mass spectrometry(UPLC-MS). Orthogonal partial least squares discriminant analysis(OPLS-DA) was performed to compare the differences of metabolomics among different region, sex, and age.Results:Significant differences on metabolomics profiles were identified among region, sex, and age. A total of 114 region-related metabolites were spotted, including 53 metabolites that involved in human metabolic pathways, mainly peptides(20 metabolites) and glycerophospholipid metabolism-related(14 metabolites). Fifty-nine metabolites were pinned down to be sex-related, among which cotinine was significant in all 7 provinces. Age-related metabolites were only found in Shaanxi and Hainan, with 22 metabolites were recognized.Conclusion:Serum metabolomics varies by geographic regions, sex, and age. When metabolomics is applied for diagnosis or biomarker screening in various studies, it shall take into consideration of setting tailored references.

Objective:To observe the effects of the Chinese medicine prescription Xiao-Cheng-Qi decoction (XCQD) on acute brain edema and inflammatory factors in rats with severe traumatic brain injury (sTBI).Methods:A total of 108 male Sprague-Dawley (SD) rats were divided into control group, sham operation group, sTBI model group, and XCQD low, medium, high dose groups by random number table method, with 18 rats in each group. sTBI rat model was prepared according to the modified Freeney method. At 6 hours after injury, the XCQD low, medium, and high dose groups were given XCQD 1.80, 2.78, and 4.59 g/kg by gavage, respectively, and the other three groups were given the same amount of normal saline, once a day for 3 days. After 3 days of injury, rats in each group were sacrificed after the modified neurologic severity score (mNSS) assessed. Pathological changes of brain tissue were observed under light microscope after hematoxylin eosin (HE) staining, water content of brain tissue was measured by dry-wet specific gravity method, and the expressions of aquaporin 4 (AQP4), tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in brain tissue were detected by Western blotting. Serum TNF-α and IL-1β levels were detected by enzyme linked immunosorbent assay (ELISA).Results:Compared with the normal group, the mNSS score of rats increased significantly, the structure of brain tissue was disordered, and pathological changes appeared such as inflammation, edema, pyknosis of nerve nuclei, water content, the protein expressions of AQP4, TNF-α and IL-1β in brain tissue, and the contents of TNF-α, IL-1β in serum were significantly increased. After XCQD intervention, the above indexes were significantly improved. Compared with sTBI model group, the mNSS score of XCQD medium and high dose groups significantly decreased (6.94±1.16, 6.88±1.02 vs. 8.61±1.09, both P < 0.05), and the pathological changes such as brain edema and inflammation were alleviated. Brain tissue water content, AQP4 protein expression and contents of serum TNF-α, IL-1β in XCQD low, medium, and high dose groups significantly decreased compared with sTBI model group [brain tissue water content: (78.25±0.71)%, (77.62±0.44)%, (76.70±0.74)% vs. (80.08±0.66)%; the expression of brain AQP4 protein (AQP4/β-actin): 0.86±0.13, 0.84±0.22, 0.65±0.13 vs. 1.08±0.14; serum TNF-α (ng/L): 106.34±15.07, 95.75±17.26, 89.00±17.36 vs. 141.96±29.47; serum IL-1β (ng/L): 90.41±12.88, 72.82±13.51, 71.32±16.79 vs. 128.57±22.56, respectively, all P < 0.05]. The protein expressions of TNF-α,IL-1β in brain tissue of XCQD medium and high dose groups also significantly decreased compared with sTBI model group [TNF-α (TNF-α/β-actin): 0.90±0.24, 0.79±0.35 vs. 1.17±0.15; IL-1β (IL-1β/β-actin): 0.91±0.21, 0.68±0.28 vs. 1.23±0.08, respectively, all P < 0.05]. Brain tissue water content, the expression of brain AQP4 protein, the levels of brain tissue and serum IL-1β in XCQD high dose group improved more significant than those of XCQD low dose group. Conclusions:XCQD can alleviate the acute brain edema in sTBI rats, and it is dose-dependent. The mechanism may be relevant to reduce the secondary inflammatory response of sTBI by inhibiting the expression of inflammatory factors TNF-α and IL-1β.

Ulcerative Colitis (UC) has been reported to be related to Porphyromonas gingivalis (P. gingivalis). Porphyromonas gingivalis peptidylarginine deiminase (PPAD), a virulence factor released by P. gingivalis, is known to induce inflammatory responses. To explore the pathological relationships between PPAD and UC, we used homologous recombination technology to construct a P. gingivalis strain in which the PPAD gene was deleted (Δppad) and a Δppad strain in which the PPAD gene was restored (comΔppad). C57BL/6 mice were orally gavaged with saline, P. gingivalis, Δppad, or comΔppad twice a week for the entire 40 days (days 0-40), and then, UC was induced by dextran sodium sulfate (DSS) solution for 10 days (days 31-40). P. gingivalis and comΔppad exacerbated DDS-induced colitis, which was determined by assessing the parameters of colon length, disease activity index, and histological activity index, but Δppad failed to exacerbate DDS-induced colitis. Flow cytometry and ELISA revealed that compared with Δppad, P. gingivalis, and comΔppad increased T helper 17 (Th17) cell numbers and interleukin (IL)-17 production but decreased regulatory T cells (Tregs) numbers and IL-10 production in the spleens of mice with UC. We also cocultured P. gingivalis, Δppad, or comΔppad with T lymphocytes in vitro and found that P. gingivalis and comΔppad significantly increased Th17 cell numbers and decreased Treg cell numbers. Immunofluorescence staining of colon tissue paraffin sections also confirmed these results. The results suggested that P. gingivalis exacerbated the severity of UC in part via PPAD.
Animals ; Colitis, Ulcerative/microbiology* ; Mice ; Mice, Inbred C57BL ; Porphyromonas gingivalis/pathogenicity* ; Protein-Arginine Deiminases ; Virulence Factors

Objective:To study the application and effect of hybrid teaching based on the rain classroom in the teaching of periodontology for undergraduates.Methods:Sixty-four stomatological undergraduates in Phase 101 were selected as experimental group, and 62 in Phase 100 as control group. The hybrid teaching based on the rain classroom were conducted in experimental group, and mind mapping were used before and after class to help students understand and expand the content of class. The traditional teaching was adopted in the control group. SPSS 17.0 was applied to compare the differences between the written test scores and teaching satisfaction evaluation of the two groups. The learning behavior data of the students in experimental group before, during and after class were collected and analyzed.Results:The score of the written test in the experimental group were significantly better than that in the control group ( t=2.97, P<0.05). The result of questionnaire showed that the experimental group was significantly more satisfied with the improvement of lecture efficiency ( t=2.71, P<0.05) and self-learning ability ( t=4.35, P<0.05). The majority (82.55%) of students in the experimental group are in favor of the rain classroom; 89.06% of students finished the preview before class, and 67.19% of them completed the review after class. Conclusion:The hybrid teaching based on the rain classroom can be applied to the teaching of periodontology, which can effectively improve the learning quality and initiative of the students.

Objective:To observe the dynamic of neurological severity scores (NSS) and the expressions of Wnt/β-catenin signaling pathway, brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) in rats with severe traumatic brain injury (sTBI), and to explore the effect of Huoxue Huayu decoction.Methods:A total of 126 Sprague-Dawley (SD) rats were randomly divided into seven groups by random number table with 18 rats in each group, namely control group (normal saline 2 kg/L), model group (normal saline 2 kg/L), brain protolysate group (BP group, 5.6 g/kg), Taohong Siwu decoction group (TH group, 10.2 g/kg), Xuefu Zhuyu decoction group (XF group, 15.6 g/kg), Tongqiao Huoxue decoction group (TQ group, 9.6 g/kg) and Buyang Huanwu decoction group (BY group, 28.7 g/kg). The sTBI rat model was reproduced by modified Feeney free fall method, and the rats in the control group were not treated with trauma. The rats in each group were intragastrical administered with corresponding drugs at 6 hours after injury, and the NSS scores were evaluated on the 1st, 3rd and 7th days after injury. After the hippocampus was harvested, the mRNA expressions of Wnt3a and β-catenin were detected by reverse transcription-polymerase chain reaction (RT-PCR), and the positive expressions of BDNF and NGF were detected by immunohistochemistry.Results:Compared with the control group, the rats in the model group showed obvious symptoms of craniocerebral injury at 1 day after injury, which was manifested as significantly increased NSS score, up-regulated mRNA expressions of Wnt3a and β-catenin, and increased positive expressions of BDNF and NGF, which indicated that the sTBI rat model was successfully prepared and presented a certain self-repair ability with the extension of time. Compared with the model group, NSS scores in the XF group, TQ group and BY group significantly decreased at 1 day after injury (6.6±1.5, 6.1±2.0, 5.7±2.4 vs. 9.4±1.5, all P < 0.05); however, the NSS scores in the BP group and TH group decreased significantly at 7 days after injury, and the NSS scores in the TQ group and BY group decreased more significantly than those in other drug groups. Compared with the model group, mRNA expressions of Wnt3a and β-catenin in the hippocampus of the BP group increased significantly at 1 day and 3 days after injury, respectively, and continued to increase with the extension of time. The mRNA expression levels of Wnt3a and β-catenin in the four groups of Huoxue Huayu decoction fluctuated to varying degrees from 1 day to 3 days after injury, but they were significantly higher than those in the model group at 7 days after injury, and the increase was more significant in the BY group [Wnt3a mRNA (2 -ΔΔCt): 154.7±4.1 vs. 17.4±1.0, β-catenin mRNA (2 -ΔΔCt): 17.05±0.45 vs. 2.74±0.13, both P < 0.05], and the second was the TQ group [Wnt3a mRNA (2 -ΔΔCt): 126.6±2.8 vs. 17.4±1.0, β-catenin mRNA (2 -ΔΔCt): 8.70±1.19 vs. 2.74±0.13, both P < 0.05]. Compared with the model group, the positive expressions of BDNF and NGF in the BP group increased significantly at 1 day after injury, but decreased after 3 days after peak. The positive expressions of BDNF and NGF in the four Huoxue Huayu decoction groups fluctuated to varying degrees from 1 day to 3 days after injury, but they were significantly higher than those in the model group at 7 days after injury, among which, the positive expressions of BDNF and NGF in the TQ group and BY group were significantly higher than those in the model group at 1 day after injury [BDNF positive cells (cells/MP): 56.4±6.2, 61.6±7.0 vs. 37.4±2.0, NGF positive cells (cells/MP): 58.4±5.0, 62.4±4.4 vs. 53.4±3.6, all P < 0.05], the increase amplitude at 7 days after injury was more significant than those in the other groups. Conclusions:Taohong Siwu decoction, Xuefu Zhuyu decoction, Tongqiao Huoxue decoction and Buyang Huanwu decoction have curative effect on the nerve regeneration and repair of rats with sTBI at acute stage, but the intensity of the effect is different. Buyang Huanwu decoction and Tongqiao Huoxue decoction have a fast and better effect.

To summarize the breeding and identification of Wuzhishan miniature pig models with α-l,3-galactosyltransferase (GGTA1) gene-knockout (GTKO).Methods The breeding and reproduction perform of GTKO Wuzhishan miniature pigs were assessed and the quantity of piglets was counted.The GTKO Wuzhishan miniature pig models with GGTA1gene knockout were validated by polymerase chain reaction(PCR).The αGal phenotype of peripheral blood mononuclear cells (PBMC) in human,wild-type Wuzhishan miniature pigs and GTKO Wuzhishan miniature pigs was detected by fluorescent microscope and flow cytometry.Routine blood test parameters were statistically compared between the GTKO and wild-type Wuzhishan miniature pigs.Results The inheritance of GGTA1 genotype complied with Mendel's law.Flow cytometry detected no fluorescent expression of PBMC in GGTA1-/-pig models,which were consistent with the genotype identification results.The mean piglets of the primiparous GTKO Wuzhishan miniature pigs were (6.8±1.8) and (8.3±2.2) for the multiparous Wuzhishan miniature pigs.No statistical significance was noted in routine blood test parameters between the GTKO and wild-type Wuzhishan miniature pigs (all P＞0.05).Conclusions Stable inheritance and normal reproductive capacity are observed in two generations of Wuzhishan miniature pigs continuously.GTKO Wuzhishan miniature pig is a reliable donor for heterogeneous organ transplantation.

Metabolic syndrome (MS)is a clinical syndrome featured by glucose and lipid metabolic disorders,hyper- tension and obesity etc.,whose key links are insulin resistance and abdominal obesity.MS is closely related with cor- onary atherosclerosis,every link of MS can lead to occurrence and development of atherosclerosis.The present arti- cle mainly stated research progress of the correlation between MS and coronary atherosclerosis in recent years.

OBJECTIVEIn order to determine the function of PG0839 gene from Porphyromonas gingivalis (P. gingivalis) W83 strains, we intended to create a mutant in the PG0839 gene by homologous recombination.

METHODS1 584bp PG0839 gene fragment was amplified, digested by BamH I and EcoR I, purified and ligated to pUC19. The recombinant plasmid was designated as pPG0839-1. The erm cassette (2 101 bp) was inserted into the EcoR V restriction site of the PG0839 gene. The resultant recombinant plasmid, pPG0839-2, was used as a donor in the electroporation of P. gingivalis W83. After electroporated and selected on erythromycin brain heart infusion plates, a single colony was collected and designated as PG0839 gene-defective mutant.

RESULTSA mutant in PG0839 gene was created by insertional inactivation, and inactivation of PG0839 gene was confirmed by restriction endonuclease digestive, sequencing, polymerase chain reaction (PCR) and reverse transcription PCR.

CONCLUSIONA PG0839 gene-defective mutant was created successfully.

Base Sequence ; Genes, Bacterial ; Polymerase Chain Reaction ; Porphyromonas gingivalis

BACKGROUND: Polyethylene glycol-polyethyleneimine/ferroso-ferric oxide (PEG-PEI/Fe_3O_4) was selected as drug carders in tumor treatment, which can increase drug loading capacity and targeting capacity.OBJECTIVE: To test the toxicity of PEG-PEI/Fe_3O_4 nano-magnetic fluid in vitro and in vivo. METHODS: When the prepared PEG-PEI/Fe_3O_4 nano-magnetic fluid reached nano level, 7702 and HpG2 cell lines were filtrated and diluted in 5-20 multiple, and detected by in vitro MTT toxicity test assay; in vivo hemolysis test and micronucleus test was used to test the toxicity and biocompatibility.RESULTS AND CONCLUSION: MTT assay results indicated that the toxicity grade of PEG-PEI/Fe_3O_4 nano-magnetic fluid to 7702 cell line was 0-1, which was harmless to natural hepatic cells; however, PEG-PEI/Fe304 nano-magnetic fluid had slight bystander restraining effect to HpG_2 cell line. Maximum hemolysis rate of the matedel was 0.372%, which was far less than 5%. The micronucieus test result indicated that PEG-PEI/Fe_3O_4 nano-magnetic fluid had no teratogenicity or mutagenicity.

OBJECTIVETo study the relationship between donor TNFalpha - 308 (G/A) genotype and recipient acute graft versus host disease (aGVHD).

METHODSTNFalpha - 308 (G/A) and TNFalpha - 308 (G/G) genotypes were analyzed by DHPLC and DNA sequence in twenty-one degree III/IV (III/IV) aGVHD patients, and twenty-eight degree 0/I (0/I) aGVHD patients. Serum TNFalpha levels were determined by ELISA.

RESULTSThe frequency of TNFalpha - 308 (G/A) genotype was significantly higher in III/IV aGVHD patients than in 0/I aGVHD patients (8/21 vs 1/28) (P < 0.05); and so did for higher serum TNFalpha levels (P < 0.05). TNFalpha levels were higher in III/IV aGVHD patients than in 0/I aGVHD patients in TNFalpha - 308 (G/G) patients (P < 0.01).

CONCLUSIONTNFalpha - 308 (G/A) is positively related to serum TNFalpha levels and most likely contributes to high risk for III/IV aGVHD.

Acute Disease ; Base Sequence ; Blood Donors ; Chromatography, High Pressure Liquid ; methods ; DNA ; chemistry ; genetics ; Enzyme-Linked Immunosorbent Assay ; Gene Frequency ; Genotype ; Graft vs Host Disease ; blood ; genetics ; Hematopoietic Stem Cell Transplantation ; Humans ; Molecular Sequence Data ; Sequence Analysis, DNA ; Tumor Necrosis Factor-alpha ; genetics ; metabolism