Abstract:Objective：To explore the effects of glutamine synthetase (GS) inhibitor on the homoeostasis of amino acid neurotransmitters and monoamine neurotransmitters in the striatum of mice. Methods：Three-month-old C57BL/6J mice were intraperitoneally injected with glutamine synthetase inhibitor-methionine sulfoximine (MSO) for 2 weeks (0.1 mL/10 g，qod). The protein levels of GS and glutamate transporters were detected by Western blot. The concentrations of amino acid neurotransmitters (glutamic acid and glutamine) and monoamine neurotransmitters (dopamine and 5-hydroxytryptamine) in the striatum were measured by high performance liquid chromatography. Results：Compared with the saline control group，MSO effectively inhibited GS protein expression and increased the levels of glutamate transporters (glutamate transporter 1 and glutamate aspartate transporter) in the striatum. Mice with MSO injection exhibited lower levels of glutamate，glutamine and aspartic acid，and higher levels of γ-aminobutyric acid and serine，but showed no change in the concentrations of monoamine neurotransmitters. Conclusion：Systematical inhibition of glutamine synthetase could affect the homoeostasis of metabolism of amino acid neurotransmitters in the brain of mouse，and lead to the decrease of the excitatory neurotransmitter glutamate level and the increase of inhibitory neurotransmitter γ-aminobutyric acid level.

Abstract:Objective:To in vestigate the method of culturing neural stem cell s(NSCs) in vitro and measurement of n6/n3 ratios and DHA contents in these NSCs cultured from subventricular zone in adult mice. Methods：Subventricular zone was isolated from the brains of 8～10 weeks old C57BL/6 mice, and the NSCs were cultured in floating in vitro. Some marker genes of NSCs and differentiated cells were detected by immunofluorescendce; polyunsaturated fatty acids and n6/n3 ratios were measured by gas chromatography. Results：NSCs proliferated continuously in vitro, most cells were Nestin positive. NeuN，TuJ1，GFAP and O4 positive cells were detected by immunofluorescence in differentiated neural cells. Polyunsaturated fatty acids were richer in NSCs. Conclusion：NSCs cultured from subventricular zone in adult mice have proliferation and self-renewal abilities. n3 polyunsaturated fatty acids and DHA in NSCs are significantly higher than those in tails.

Abstract:Objective：To investigate the inhibitory effects of apigenin on interleukin 6 (IL-6)-induced delay in neutrophil apoptosis and its underlying mechanisms.Methods：Neutrophils(Neu) isolated from healthy volunteers were randomly divided into the control group，apigenin group，IL-6 group，IL-6+apigenin group，QVD+IL-6+apigenin group，and z-VAD+IL-6+apigenin group.Neu apoptosis was detected by flow cytometry.The protein expression of cleaved caspase-3 was determined by Western blotting analysis.Results：Compared with the control group，apigenin significantly induced Neu apoptosis (P＜0.05)，and IL-6 significantly inhibited Neu apoptosis (P＜0.05).Apigenin partially reversed the anti-apoptotic effect of IL-6 on Neu (P＜0.05).Compared with the IL-6 group，cleaved caspase-3 expression was significantly increased in the IL-6+apigenin group.Moreover，caspase inhibitors QVD-Oph and z-VAD reduced the levels of cleaved caspase-3 and abolished the ability of apigenin to counteract IL-6-induced delay in Neu apoptosis.Conclusion：Apigenin overrides IL-6-induced delayed apoptosis of Neu in a caspase-dependent manner，which provides a promising target for future therapy of neutrophilic inflammatory airway disorders.

Abstract:Objective：To study the role of IKKα in macrophages in renal inflammation after renal ischemia-reperfusion injury(IRI)in mice model.Methods：A total of 20 C57BL/6 wild type (WT) mice and 20 IKKαMKO (KO) mice (healthy，male，and aged 8～10 weeks) were randomly divided into the control (Sham) group and the renal IRI group，respectively.The morphology change of renal tissue and the infiltration of inflammatory cells were observed by hematoxylin and eosin (HE) staining.The expressions of interleukin 10(IL-10)，interleukin 6(IL-6)，Ki67，CD68，iNOS，and Arg-1 were detected by immunohistochemistry.The protein expressions of IL-10 and IL-6 were detected by Western blotting.Results：Compared with the Sham group，obvious renal tissue injury and increasing inflammation infiltration were observed in the IRI group by HE staining. Immunohistochemical results on expressions of both IL-10 and IL-6 were increased after renal IRI occurred，IL-10 increased while IL-6 decreased by time.Compared with the WT-IRI group，the renal pathological damage was more severe (P＜0.01)，the expressions of IL-6，CD68 and iNOS were significantly increased (P＜0.01)，but the expressions of IL-10 (P＜0.01) and Ki67 (P＜0.05) were distinctly decreased in the KO-IRI group.Conclusion：IKKα knockout in macrophages aggravates renal ischemia-reperfusion injury and inhibits renal repair in mice model，which can be related to intensifying the early infiltration and inflammatory reaction of macrophages(M1 mainly).

Abstract:Objective：To detect the expression of interleukin(IL)-33 in renal cell carcinoma(RCC) and its effect on invasion of human 786-0 cells.Methods：IL-33 protein in 34 patients with different grades of RCC tissues and para-adjacent tissues was detected with immunohistochemistry assay.Transwell invasion assay was used to investigate the effect of IL-33 on invasion of 786-0 cells.Western blot assay was conducted to detect the markers of epithelial-mesenchymal transition(EMT) in 786-0 cells after IL-33 treatment.Results：IL-33 protein expression was significantly higher in RCC tissues compared to para-adjacent renal tissues(P＜0.001).IL-33 enhanced 786-0 cells invasion in vitro.Moreover，the expression of E-cadherin was significantly decreased while the expression of vimentin was significantly increased after IL-33 treatment.Conclusion：IL-33 is overexpressed in RCC tissues and may contribute to renal cancer cells invasion by inducing EMT.

Abstract:Objective：To observe the effects of astaxanthin on paraquat poisoning-induced pulmonary fibrosis in SD rats， and explore the mechanism of pulmonary fibrosis. Methods：The rats were randomly divided into three groups， including the normal control group， the PQ model group， and the astaxanthin group. The PQ group and the astaxanthin group were intraperitoneal injected with PQ. In the next day， the rats were treated by intragastric administration once a day. The control group and the PQ group were treated by the same dosage of solvent． On day 7， day 14， day 28 after treatment， the rats in each group were sacrificed. The levels of hydroxyproline （HYP） and TGF-β1 were measured. The levels of matrix metalloproteinase （MMP） 1， MMP2， MMP9 and tissue inhibitor of matrix metalloproteinase1 （TIMP1） were determined in the serum of rat by ELISA， and the pathologic changes of lung tissues were observed by HE staining and Masson staining. Results：①HE staining and Masson staining results showed that the degree of fibrosis in the astaxanthin group was significantly lower than that of the PQ group. ②The hydroxyproline content of model group was significantly higher than that of the normal group （P<0.05）. In the intervention of astaxanthin， the level of hydroxyproline in rat was decreased compared with the model group （P<0.05）. ③TGF-β1 was expressed in the PQ model group. After intervention of astaxanthin， the expression of TGF-β1 was decreased， especially at day 28， and the difference was statistically significant （P<0.05）. ④The expression levels of MMP1， MMP2 and MMP9 in the model group and the astaxanthin group were higher than those in the normal control group. The expressions of MMP1， MMP2， MMP9 after PQ poisoning were gradually increased， reached the highest point in day 14， then the expressions in day 14 began to decline but still higher than those of the control group. Compared with the model group， the contents of MMP9 and MMP2 in the astaxanthin group were decreased， and the difference was statistically significant （P<0.05） in the day 14 and day 28. TIMP1 had no significant difference between the model group and the control group during the intervention period （P>0.05）. Conclusion：Astaxanthin can reduce the content of hydroxyproline， MMP9， and MMP2， inhibit the secretion of TGF-β1 in lung tissue， and decrease the collagen deposition in lung tissue of rats with pulmonary fibrosis， further reduce PQ-induced pulmonary fibrosis in rats.

Abstract:Objective：To observe effects of metformin on proliferation and related gene expression of human pancreatic cancer cell line Patu8988 treated by diabetic serum.Methods：Patu8988 cells were cultured with diabetic serum，or healthy human serum as control.The two groups were treated with different concentrations of metformin (2.5，5.0，10.0，20.0，and 40.0 mmol/L.The proliferation was measured by cell counting kit-8 (CCK-8) assay，and the expressions of matrix metalloproteinase-2 (MMP-2)，matrix metalloproteinase-8 (MMP-8) and matrix metalloproteinase-11 (MMP-11) were examined by real-time PCR.Results：Compared with the control group，the proliferation of Patu8988 was promoted after treatment with diabetic serum (0.74±0.03 vs.0.62±0.02，P＜0.01)：Metformin inhibited the proliferation of Patu8988 treated with diabetic serum (P＜0.01)，and the expressions of MMP-2，MMP-8 and MMP-11 were downregulated.Conclusion：Metformin inhibits the proliferation of cancer cells induced by the serum from patients with diabetes，which may be related to the down regulation of MMP-2，MMP-8, and MMP-11.

Abstract:Objective:To construct a lentiviral vector containing Mxi1 gene and observe the effect of Mxi1 on proliferation and apoptosis of SGC-7901 cells. Methods:The Mxi1 gene was cloned into lentiviral expression vector by recombining DNA technology.The recombinant plasmid was identified by restriction enzyme digestion and gene sequencing，and then cotransfected 293T cells with the auxiliary packaging components plasmids to obtain recombinant lentivirus containing Mxi1 gene.SGC-7901 cells were infected with the recombinant lentivirus，and the infection efficiency was observed under fluorescence microscope.Expressions of Mxi1，cyclinB1 and caspase-8 gene were identified by RT-PCR and expression of Mxi1 protein was identified by Western blot. The proliferation and apoptosis of SGC-7901 cells were examined by CCK-8 and flow cytometry，respectively.Results:The Mxi1 gene was successfully cloned to lentiviral，expressions of Mxi1 gene and protein were confirmed by RT-PCR and Western blot.Mxi1-overexpressed SGC-7901 cells expressed less cyclinB1 and more caspase-8 compared to blank control cells and empty vector-overexpressed cells confinned by RT-PCR.CCK-8 and flow cytometry experimental results showed that the proliferation ability of Mxi1-overexpressed SGC-7901 cells was deteriorated significantly compared to blank control cells and empty vector-overexpressed cells，and the apoptosis rate of Mxi1-overexpressed cells was higher than that of blank control cells and empty vector-overexpressed cells.Conclusion:The Mxi1 recombinant lentiviral vector had been successfully constructed and effectively transfected SGC-7901 cells.Mxi1 could inhibit the proliferation and promote the apoptosis of SGC-7901 cells.

Abstract:Objective：To investigate whether the α7 nicotinic acetylcholine receptors agonist GTS-21 can ameliorate dextran sulfate sodium(DSS)-induced colitis and its potential mechanism. Methods：Male BALB/c mice(8-10 weeks old) were randomly divided into 3 groups，including the control group，the DSS-induced group，and the GTS-21 treatment group(n=8，each group). DSS model was performed with 3.5% DSS free drink，the GTS-21 treatment group was performed with 3.5% DSS free drink+GTS-21 10 mg/(kg·d)，ip，for 7 d. The control group was given normal saline，and DAI score was given to each group for every day. At day 8，all mice were sacrificed，the changes of the colonic mucosa were observed，the length and wet weight were measured，and the histological inflammation(HI) score was assessed by HE staining. The levels of cytokines were detected by ELISA. Results：①Compared with the control mice，DAI score was increased［(1.51±0.10)vs.0，n=8，P＜0.001］，colon length was shortened［(8.22±0.37)cm vs.(11.65±0.30)cm，n=8，P＜0.001］ and HI score was increased［(20.5±3.9)vs.(0.9±0.4)，n=8，P＜0.001］in mice-induced DSS，which showed that the model was successfully established.②Compared with DSS-induced mice，DAI score was decreased［(0.25±0.10)vs.(1.51±0.10)，n=8，P＜0.001］，shortened colon length was improved［(9.42±0.32)cm vs.(8.22±0.37)cm，n=8，P＜0.05］ and HI score was decreased［(7.5±2.0)vs.(20.5± 3.9)，n=8，P＜0.01］ in mice with administration of GTS-21，which suggested that GTS-21 improved DSS-induced colitis in mice.③The level of CXCL9/Mig in DSS-induced mice was increased，while it was significantly decreased in mice with administration of GTS-21. In addition，tumor necrosis factor α(TNF-α)，interleukin 1β(IL-1β) and interferon γ(IFN-γ) were significantly increased. ④CXCL9/Mig with the most obvious change detected by further ELISA showed that serum CXCL9/Mig increased significantly in the DSS group(P＜0.05)，and after administration of GTS-21，serum CXCL9/Mig was decreased(P＜0.05). Conclusion：α7nAChR agonist GTS-21 can attenuate the intestinal inflammation in DSS-induced mice，which may be due to reduction of CXCL9/Mig(a chemotactic factor)，and then reduction of inflammatory cell aggregation.

Abstract:Objective: To observe the effects on proliferation and apoptosis in human umbilieal vein endothelial cells （HUVECs） cultured with the serum of the children with coronary artery lesions of Kawasaki disease， in order to further understand the possible mechanism of vascular endothelial injury from Kawasaki disease. Methods: HUVECs were cultured and divided into 4 groups， including the normal serum group， the general fever group， the non-CALs group， and the CALs group. The proliferation was detected by MTT test and the apoptosis was detected by flow cytometry （FCM）. Results: The OD value of the HUVECs detected by MTT test after being cultured with the serum of the children with Kawasaki disease including the Non-CALs group and the CALs group （0.492±0.033 and 0.395±0.031， respectively） were lower than that of the general fever group and the normal serum group （0.592±0.033 and 0.612±0.036， respectively） （P<0.01）. The apoptosis of HUVECs detected by FCM after being cultured with the serum of the children with Kawasaki disease was higher than that of the other two groups （P<0.01）， and the apoptosis of CALs group was higher than that of the Non-CALs group. Conclusion: The serum from children with coronary artery lesions of Kawasaki disease could inhibit the proliferation of HUVECs and induce the apoptosis。

Abstract:Objectives：To learn the factors related to energy consumption among residents aged from 40～60 years in Nanjing. Methods：A total of 110 men and women were recruited in Nanjing，their height and body weight were measured，and then body mass index (BMI) and body surface area (BSA) were calculated. The body composition information was recorded according to the dual energy X-ray body scanner. The energy consumption was respectively observed for flat-lying，exercising，internet browsing，sleeping，and rest metabolism during 11 hours they lived in the metabolism chamber. The relationship was analyzed between the energy consumption of different activity states and age，gender，body weight，height，BMI or body composition. Results：①The basal metabolism of male and female subjects was (2 036.25 ± 226.45) kcal/d，(1 673.21 ± 173.37) kcal/d，respectively，the difference was significant (P<0.01)；There were no significant differences in basal metabolism between subjects of different age groups (P>0.05). The energy consumption of women at different activity states was significantly higher than that of male (P<0.01) when energy expenditure (EE)/fat-free mass (FFM)；However，there were no significant differences in energy consumption between subjects at different ages when EE/FFM (P>0.05). ②The energy consumption of male and female subjects was significantly correlated with FFM，body weight，BSA，BMI，and body fat (P<0.01). According to the multiple regression analysis，it was found that the energy consumption under different active states was affected by gender，body fat，and FFM (P<0.05). Conclusion：The energy consumption of different activity states is mainly related to the FFM and body fat among the residents aged from 40～60 years in Nanjing.

Abstract:Objective：The present study is aimed to evaluate the quality of two clinical practice guidelines on stomatology published lately in China and overseas so that the limitations of development of clinical practive guideline in China could be emphasized，which will be helpful to improve the methodological quality to develope evidence-based guide line and to update the current guidelines in the future. Methods：Guidelines on stomatology published from 2014 to 2016 were electronically searched in PubMed，Wanfang，CNKI and databases of National Guideline Clearing house. One guideline on stomatology developed in China and another one developed overseas were selected. The quality was evaluated by the AGREE II tool and then descriptive analysis was performed. Results：Only one clinical practice guideline on stomatology was published in China. i. e. Guidelines for the use of microscopes in endodontics in 2016. Another clinical practice guideline(Evidence-based clinical practice guideline for the use of pit-and-fissure sealants) developed in USA was selected. Based on 23 items organized into the original 6 quality domains，the selected foreign guideline was deemed to conform to 22 items except for 1 item. The selected domestic guideline did only conform to 1 item. Conclusion：Compared with the Evidence-based clinical practice guideline for the use of pit-and-fissure sealants developed in USA，the domestic clinical practice guideline has not yet conformed to the procedure to develop the evidence-based clinical practice guideline，which indicates that domestic stomatology experts should emphasize the transparency and methodology rigor when they develop new clinical practice guidelines.

Abstract:Objective：To evaluate the reliability of cone-beam computed tomography (CBCT) on identification of facial symmetry landmarks，and provide evidences for the measurement and analysis of three-dimensional facial symmetry. Methods：The CBCT images of 57 orthodontic patients were measured by multi planar reconstruction (sagittal，axial and coronal to the point) by using mimics 10.0 software to calibrate 28 facial symmetry related landmarks. There were 2 observers in this study，each observer had repeated 3 times with 2 weeks and 4 weeks intervals，respectively. The intra- and inter-class correlation coefficient (ICC) of each dimension coordinate value of each landmark was calculated to evaluate the reliability of the landmarks for intra-observer and inter-observer. Results：The ICCs of Ba，PNS，Cg，N，Antcr (L，R)，D'(L，R)，K (L，R)，Maz (L，R)，Or (L，R)，Zyg (L，R)，Cd (L，R) were with high reliability，and could be used for the analysis of CBCT three-dimensional cephalometry of facial symmetry. Conclusion：In this study，there are 19 in 28 facial symmetry related landmarks with high reliability，which are of great value in clinical practice.

Abstract:Objective：To determine the contamination of Pseudomonasa spp. in dental unit waterlines(DUWLs) and water samples using bacterial culture technique as well as Miseq sequencing technology.Methods：Water samples were collected from 25 dental units(DU) at the Affiliated Stomatology Hospital of Nanjing Medical University.Before the opening of the clinic，a water sample of 400 mL was collected from each DU using triple change syringe.Each sample was divided into two parts.One part of the water samples was used for bacterial culture.The other one was used for extraction of the total bacterial DNA，which was subjected to PCR amplification using bacterial universal primers.The PCR products of each sample were constructed using Miseq sequencing technology to complete the sequencing analysis and biological information analysis.Results：No Pseudomonas spp. was detected by bacterial culture in 25 samples under the condition of different concentrations.The metagenomic libraries were successfully constructed by 16 samples.The detection rate of Pseudomonas spp. was 100%.The relative abundance of Pseudomonas spp. in 16 samples was accounted for(3.02± 2.12)%.Conclusion：Low level of contamination of Pseudomonas spp.existed in DUWLs. Most clones of the Pseudomonas spp. in DUWLs were dead or in viable but nonculturable(VBNC) condition.Thus，Pseudomonas spp. presented a low biohazard risk in DUWLs.When compared to bacterial culture，Miseq sequencing technology is more accurate in detecting particular pathogens. However，Miseq sequencing technology has limitation on identification of viable bacterial from dead ones，which may lead to false positive results.It needs further researches using high-throughput sequencing technology for selectively detecting viable pathogens.

Abstract:Objective：To evaluate the diagnostic value of serum mesothelin level on ovarian cancer. Methods：Relevant studies were identified by systematically searching PubMed，Embase，Web of Science，CBMdisc，WANFANG，CNKI and so on，articles related to the diagnostic value of serum mesothelin on ovarian cancer were selected. Data analysis was conducted by RevMan 5.3，Stata 12.0 and Meta-Disc1.4. Results：Nine studies with a total of 451 ovarian cancer cases and 617 controls were included. The results of meta-analysis showed that sensitivity and specificity were 0.80 (95%CI：0.76～0.83) and 0.81 95%CI：0.78～0.84)，respectively;positive likelihood ratio and negative likelihood ratio were 6.93 (95%CI：3.32～14.46) and 0.24 (95%CI：0.16～0.35)，respectively;diagnostic odds ratio was 45.69 (95%CI：17.49～119.35);the area under curve of the summary receiver operating characteristic(SROC) was 0.90，Q=0.83. Conclusion：Mesothelin shows a high specificity and a low sensitivity in the diagnosis of ovarian cancer，and it can be used as an effective way for diagnosis of ovarian cancer combined detection with other tumor markers.