Nitric oxide (NO), which is involved in the regulation of the cardiovascular system, nervous system, immune system, reproductive system, digestive system and other physiological activities, is an important biological substance with activity. Under normal physiological conditions, neuronal nitric oxide synthase (nNOS) can precisely regulate the nervous system NO production, release, diffusion and inactivation processes. But an excess of NO associates with the development of cerebral ischemia, Alzheimer's and Parkinson's psychosis nervous system diseases, while inhibition of nNOS activity can regulate the content of NO in vivo, and produce a therapeutic effect on some of the nervous system diseases. This review mainly describes the structure and regulation of nNOS and recent developments of small molecule inhibitors of nNOS.

Objective To explore the diagnostic value of anti-cyclic citrullinated peptide (CCP) 3.1 IgG/IgA antibody detected by enzyme-linked immunosorbent assay (ELISA) in patients with rheumatoid arthritis (RA).Methods The ELISA was used to measure the anti-CCP3.1 antibody in the serum of 169 RA patients,100 patients with other rheumatic diseases (including systemic lupus erythematosus,Sjogren's syndrome and osteoarthritis) and 72 healthy controls.The diagnostic value of CCP3.1 was assessed and compared with the second generation of anti-CCP IgG (CCP2) antibody,the correlations between anti-CCP3.1 antibody and the clinical and laboratory parameters were analyzed.Two-independent samples t test,chi-square test and Spearman's correlation were adopted for statistical analysis.Results ① The average cut-off concentration of anti-CCP3.1 antibody was (1122±1429) U/ml in RA,(13±14) U/ml in other rheumatic diseases and (6±5) U/ml in healthy controls.② The area under curve of ROC for anti-CCP3.1 antibody and anti-CCP2 antibody were 0.923 and 0.936 respectively.There was no difference between the sensitivity (82％ vs 79％)and specificity (97％ vs 99％) of anti-CCP3.1 antibody and anti-CCP2 antibody.The Kappa values between anti-CCP3.1 antibody and anti-CCP2 antibody was 0.763.③ We also found that anti-CCP3.1 antibody was positive in 20％(7/35) of anti-CCP2 antibody negative,43％(18/42) of RF negative,62％(47/76) of AKA negative,71％(49/69) of APF negative and 13％ of autoantibodies negative patients,indicated that antiCCP3.1 antibody had a potential value in the diagnosis of serum negative patients with RA.④ The presence of anti-CCP3.1 antibody was correlated with RF,HRF-IgG,APF,AKA,GPI and IgA (P＜0.05),except disease activity.Conclusion The sensitivity of anti-CCP3.1 antibody is slightly higher than anti-CCP2 antibody.The Anti-CCP3.1 antibody is a very valuable parameter for the diagnosis of RA,especially in serum negative patients.

Objective To compare the condition of illness and pathological characteristics of experimental autoimmune encephalomyelitis (EAE)in C57 BL/6 mouse models induced by myelin oligodendrocyte glycoprotein 35-55 (MOG35-55)at different doses,and provide a reliable animal model for further study of multiple sclerosis(MS).Methods Male SPF-grade C57 BL/6 mice were divided randomly into four groups:normal group and three EAE model groups (MOG35-55 high-dose,middle-dose and low-dose model groups).200,100,50μg MOG35-55/mice were mixed with complete Freund's adjuvant(CFA),respectively,to prepare complete antigen in different concentrations.The mice were anesthetized and injected s.c.over flanks with the complete antigen and injected i.P.with pertussis toxin to establish immunization-induced C57BL/6 mouse-model of EAE.The mice of the normal group were injected with normal saline instead.Since the day of immunization,the incidence,body weight and neurological score of the mice were observed.The mice of different neurological scores in different periods were anesthetized and perfused with saline and followed by 4% paraformaldehyde.The brain and spinal cord of the mice were removed and fixed in the same fixative solution.The brains and spinal cords of the mice were examined by histopathology with hematoxylin-eosin(HE) staining.The mice on the 40th day were sacrificed and perfused with 2% paraformaldehyde and 2% glutaraldehyde, 1 mm~3 pieces of cerebral white matter and intumescentia lumbalis of the spinal cord were taken and ultrathin sections were prepared according to conventional techniques for electron microscopy. Results All the MOG_(35-55) in three different doses induced mouse models of EAE. The disease was with an incidence rate of 100% and a chronic monophasic course. The body weight of the mice in the three groups decreased obviously compared with those in the normal group. The maximum value of neurological score was 1.33,2.25 and 2.50 in the mice of high-, middle-and low-dose groups, respectively. The major histopathological changes observed in the brain and spinal cord of the EAE mice were different degrees of inflammatory cell infiltration around small vessels showing sleeve-like changes, dcmyelination and neuronal karyopyknosis in the acute and remission stages. The main site of the brain inflammation was in white matter around encephalocoele, and also in the DG and CA zones of hippocampus. The spinal cord inflammation was most severe in the lumbosacral region. The above mentioned pathological changes in the low-dose group were more prominent than those in the middle-dose and high-dose groups. The major ultrastructural changes were scattered around encephalocoele, interstitial edema, especially around small blood vessels, and swollen mitochondria with damaged cristae, and some karyopyknosis in vascular endothelial cells. Some tight junctions were blurred. Some dispersed lymphocytes and mononuclear cells were seen in the perivascular space. In lumbar intumescentia of the spinal cord, there were some myelin figures in the white matter myelin sheath. Some of them showed demyelization and structurtal fusion. The cytoplasmic organelles of axons were considerably reduced or even disappeared. The vascular basement membrane showed an increased thickness and focal necrosis in some areas. Conclusion The mouse models of immune-induced EAE are successfully established with MOG_(35-55), especially that induced with MOG in a dose of 50 μg. This mouse model is stable, with a high incidence and low mortality rate, and can be applied for EAE research in the future.

Objective To investigate the possible role of abnormal signal transduction in smooth muscle cell in the pathogenesis of diabetic gastroparesis. Methods Healthy male SD rats were randomly divided into 2 groups: normal,model group,diabetes was induced by Ⅳ of alloxan,50mg/kg. Gastric emptying was measured by method of nutritious semisolid paste. The level of cAMP in gastric smooth muscle cell was measured by radioimmunoassay. Gsαand GiαmRNA positive cells were quantitatively measured by in situ hybridization and computer image analysis system respectively. Results ( 1 ) Gastric emptying rate of the normal was significantly higher than that of model group (P < 0.05 ) ; (2)the level of cAMP of the normal was significantly lower than that of model group( P < 0.05 ) ; (3)the expression of GsαmRNA in the model group was significantly higher than that of the normal groups( P < 0.05 ) ; (4) there was no marked difference in the expression of GiαmRNA between 2 groups( P > 0.05). Conclusion Altered expression of GSαmRNA may play a key role in the pathogenesis of diabetic gastroparesis.

Nitric oxide (NO), which is involved in the regulation of the cardiovascular system, nervous system, immune system, reproductive system, digestive system and other physiological activities, is an important biological substance with activity. Under normal physiological conditions, neuronal nitric oxide synthase (nNOS) can precisely regulate the nervous system NO production, release, diffusion and inactivation processes. But an excess of NO associates with the development of cerebral ischemia, Alzheimer's and Parkinson's psychosis nervous system diseases, while inhibition of nNOS activity can regulate the content of NO in vivo, and produce a therapeutic effect on some of the nervous system diseases. This review mainly describes the structure and regulation of nNOS and recent developments of small molecule inhibitors of nNOS.
Alzheimer Disease ; physiopathology ; Brain Ischemia ; physiopathology ; Humans ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase Type I ; antagonists & inhibitors ; metabolism ; Parkinson Disease ; physiopathology

AIM:To investigate the autophagy induced by sepsis and acute kidney injury , and the regulation of phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway in this process.METHODS: The rats were subjected to cecal ligation and puncture ( CLP) or sham operation .Histopathologic changes of the renal tissues were examined by HE staining .Blood urea nitrogen ( BUN) and serum creatinine ( SCr) were measured by chemical colorime-try.The protein expression of microtubule-associated protein light chain 3 I/II (LC3 I/II), beclin-1 and p-Akt at different time points after CLP was detected by Western blotting .In vitro, human proximal tubular epithelial cell line HK-2 were treated with LPS to induce autophagy .The protein expression of LC 3 I/II and p-Akt in the HK-2 cells after LPS treatment at different time points and different concentrations was detected by Western blotting .These molecules in HK-2 cells and apoptosis of HK-2 cells treated with LPS plus PI3K inhibitor or Akt inhibitor were also detected .RESULTS: Compared with sham group , the severe changes of renal histopathological injuries in CLP groups were observed , the levels of BUN and SCr in CLP groups were significantly increased .LC3 I/II, beclin-1 and phosphorylation of Akt gradually increased after CLP.After treatment with LPS, the expression of p-Akt (308) in the HK-2 cells gradually increased in a dose-and time-dependent fashion.The expression of beclin-1 and p-Akt (472) reached a peak at 8 h or 10 mg/L LPS treatment.Treat-ment with PI3K or Akt inhibitor down-regulated the expression of LC3 and promoted the apoptosis of HK-2 cells.CON-CLUSION:Autophagy in the kidney is induced by sepsis and acute kidney injury .PI3/Akt signaling pathway may be in-volved in this process .

Objective:To optimize the parameters of the equation of sagittal craniofacial structures with different classes of malocclusion using genetic algorithms(GAS), and to explore the rules .Methods:A total of 240 patients with average angle malocclusion aged 8-18 years old were divided into three groups: Angle Class Ⅰ(n=79), Angle Class Ⅱ(n=76)and Angle Class Ⅲ(n=85) groups.In each group 10 cases were randomly selected as the test samples, the rest as the experimental samples.The cephalometric analysis was performed on all the patients'' cephalograms, and the results of Ba-N,Ba-A,Ba-S,S-Ptm,Ptm-A,Ba-Ar,Ar-Go,Go-PoG,Ba-PoG and N-S-Ar were analyzed by two independent samples t-test and One-Way ANOVA. The relevant influencing factors of craniofacial structures were found.The parameters of the equation was optimized to obtain the relevant equations using GAS.The predicted values of the optimized equation were compared with the measured values.Results:There were no significant differences in sex between Angle Class Ⅰ, Class Ⅱ and Class Ⅲ groups(P> 0.05);when the men and women with the same type were combined,the Ba-A,Ptm-A,Ar-Go,and Ba-PoG had statistically significant differences between Angle Class Ⅰ, Class Ⅱ, and Class Ⅲ groups (P<0.05).The correlation analysis results showed that in Angle Class Ⅰgroup:Ba-A was positively correlated with Ba-N (r=0.683),Ptm-A was positively correlated with Go-PoG (r=0.738), Ar-Go was positively correlated with Ba-PoG (r=0.833), and negatively correlated with Go-PoG (r=-0.560) and Ba-PoG was positively correlated with Go-PoG (r=0.669);in Angle class Ⅱ group,Ba-A was positively correlated with Ba-PoG and Ba-N(r=0.884,r=0.883), Ptm-A was positively correlated with Ba-A (r=0.742),Ar-Go was positively correlated with Ba-PoG (r=0.401)and negatively correlated with Go-PoG (r=-0.317) and Ba-PoG was positively correlated with Ba-A and Go-PoG(r=0.883,r=0.488);in Angle Class Ⅲ group,Ba-A was positively correlated with Ba-N and Ba-PoG(r=0.891,r=0.829),Ptm-A was positively correlated with Ba-A (r=0.807)and negatively correlated with Ba-S (r=-0.404),Ar-Go was positively correlated with S-Ptm (r=0.548) and Ba-PoG was positively correlated with Ba-A (r=0.829).The equation of sagittal craniofacial structure with different occlusal classes was established by GAS.In Angle Class Ⅰgroup:Ba-A(mm)=10.963 9+0.859 8×Ba-N,Ptm-A(mm)=6.897 6+0.557 0×Go-PoG,Ar-Go(mm)=-2.548 2+0.511 8×Ba-PoG-0.5272×Go-PoG,Ba-PoG(mm)=17.515 6+1.021 3×GO-POG;in Angle Class Ⅱ group:Ba-A(mm)=-2.121 3+0.567 6× Ba-PoG+0.513 2× Ba-N,Ptm-A(mm)=13.788 7+0.349 4×Ba-A,Ar-Go(mm)=2.447 7+0.368 8×Ba-PoG-0.427 9×Go-PoG,Ba-PoG(mm)=-7.140 2+0.751 3×Ba-A+0.295 4×Go-PoG;in Angle Class Ⅲgroup:Ba-A(mm)=3.281 0+0.545 3×Ba-N+0.394 4× Ba-PoG,Ptm-A(mm)=3.535 8+0.63 1×Ba-A-0.614 2×Ba-S,Ar-Go(mm)=-9.002 1+1.004 3×S-Ptm,Ba-PoG(mm)=-2.091 2+1.057 5×Ba-A.There were no significant differences between the predicted values of GAS and the measured data (P> 0.05), and the error was small.Conclusion: The optimal relation equation of craniofacial structure of sagittal malocclusion is established by GAS with the quantitative regularity.

Objective:To establish the quantitative relationship equation of the crantiofacial vertical points in the skeletal classⅡ malocclusion patients with various vertical types by using genetic algorithms method,and to express the measured values in the patients with different gender with the same formula.Methods:A total of 155 skeletal class Ⅱ malocclusion patients without treatment,aged from 10 to 18 years old,were selected and divided into high-angle group(n=50),average-angle group(n=58),low-angle group(n=47);5 samples were randomly selected in each group as the test samples,the rest as the experimental sample.The cephalometic radiographs were performed and measured.The relevant influencing factors of craniofacial structure were ensured.The genetic algorithm was used to optimize the equation parameters to obtain the correlation equation.The error between the predicted value and the measured value was compared.Results:The various parameters had no significant differences between different gender in high-angle,average-angle and low-angle groups(P>0.05);then the men and the women with same type were combined,most of the indicators had statistically significant differences between three groups (P<0.05).The correlation analysis results showed that there was a positive correlation between age and Ans-U1(r=0.470),there was a positive correlation between N-Me and Ans-Me(r=0.964);for Ans-U1 ,there was a positive correlation with Ans-Me(r=0.805)and negative correlation with facial angle;there was a positive correlation between N-Go and N-Me (r=0.926);for L1-Me,there was a positive correlation with Ans-Me(r=0.898)and negative correlation with the angle of Go(r=-0.468)in high-angle group. In average-angle group,there was a positive correlation between age and N-Me (r=0.531);for Ans-U1,there was a positive correlation with Ans-Me(r=0.878)and negative correlation with the facial angle(r=-0.262);for Ans-Me,there was a positive correlation with N-Me(r=0.920), negative correlation with N-Ans(r=-0.560)and negative correlation with Ar-Go(r=-0.652);for N-Go,there was a positive correlation with S-Go(r=0.867), positive correlation with N-Ans(r=0.252)and positive correlation with L1-Me(r=0.754).For S-Ar,there was a positive correlation with S-Go(r=0.671), negative correlation with Ar-Go(r=-0.250),and positive correlation with L1-Me(r=0.552).In low-angle group,for age,there was a positive correlation with S-Go(r=0.602), negative correlation with the angle of Go(r=-0.346),and positive correlation with L1-Me(r=0.576);for N-Me,there was a positive correlation with Ans-Me(r=0.869),and positive correlation with N-Go(r=0.859),and negative correlation with the facial angle(r=-0.177);for N-Ans,there was a positive correlation with N-Me(r=0.605) and negative correlation with Ans-U1(r=-0.113);for Ans-Me,there was a positive correlation with N-Me(r=0.869),positive correlation with the facial angle(r=0.070),and positive correlation with Ans-U1(r=0.785);for N-Go,there was a positive correlation with N-Me(r=0.859)and positive correlation with S-Go(r=0.829).The quantitative relationship equations of the crantiofacial vertical points in skeletal class Ⅱmalocclusion patients with various vertical types in each group were established by using genetic algorithms.In high-angle group:Age=5.883 6+0.269×Ans-U1,N-M=22.026 6+1.494 5×Ans-Me,Ans-U1=34.959 4+0.454 5×Ans-Me-0.409 7×Facial angle,N-Go=-4.588 2+0.472 4×N-Me,L1-Me=-12.590 5+0.5322×Ans-Me+0.124 3×∠Go.In average-angle group:Age=-2.944 1+0.146 8×N-Me,Ans-U1=18.917+0.476 4×Ans-Me-0.230 2×Facial angle,Ans-Me=-0.620 5+1.014 5×N-Me-0.974 1×N-Ans-0.057 6×Ar-Go,N-Go=1.631 1+0.897 8×S-Go+0.919 7×N-S+0.168 8×L1-Me,S-Ar=-1.823 1+0.845 3×S-Go-0.867 0×Ar-Go+0.202 4×L1-Me.In low-angle group:Age=11.740 6+0.152 7×S-Go-0.169 9×∠Go+0.252 5×L1-Me,N-Me=61.153 0+0.964 3×Ans-Me+0.628 6×N-Go-0.689 2×Facialangle,N-Ans=-4.949 2+1.065 8×N-Me-2.316 5×Ans-U1,Ans-Me=-25.180 0+0.418 4×N-Me+0.280 3×Facial angle+0.477 6×Ans-U1,N-Go=8.684 2+0.409 9×N-Me+0.403 3×S-Go.There was no significant difference between the predicted values of equation established with genetic algorithms and the measured data (P>0.05).Conclusion:The quantitative relationship equation of the crantiofacial vertical points in the skeletal class Ⅱmalocclusion patients with various vertical types established with genetic algorithms may show the vertical quantitative relationship and predict the growth to a certain degree.

Sepsis is a syndrome encompassing a uncertain pathobiology and threatens the health of human being. The new definition of sepsis indicates that sepsis is a life-threatening organ dysfunction caused by a dysregulated host response to infection. The exacerbation of inflammation accompanied with dysregulation of immunosuppression leads to the complicated changes of pathophysiology in sepsis. MicroRNAs (miRNAs) perform vital roles in the regulations of inflammation responses as well as organ damages associated with sepsis. The variety in the expression level of miRNA appears in the development of sepsis; monitoring and analyzing miRNA levels contributes to the diagnosis and prognosis of the disease. Controlling the expression levels of miRNAs related with sepsis is able to alleviate the organ damages and dysfunctions caused by the disease.

Objective:To elucidate the effect of hsa-microRNA-218(hsa-mir-218)on exogenous granulysin (GLS) expression in 293T cells.Methods:Total RNA was extracted from THP-1 cells induced by phorbol 12-myristate 13-acetatefor (PMA), and GLS gene was amplified by RT-PCR, and then cloned into pDsRed-Express-C1 to construct the GLS expression vector pDsRed-GLS.Then 293T cells were co-transfected with pDsRed-GLS and pGenesil-mir-218 (pGenesil-mir-control) and laser confocal microscopy was per-formed 36 h later to detect their co-expression .Total RNA and protein were extracted 48 h post transfection , and RT-PCR and Western blot were performed to detect the effect of hsa-mir-218 on exogenous GLS expression .Results:The GLS expression vector pDsRed-GLS was constructed successfully and laser confocal microscopy indicated that it was co -expressed with the interference vector .Compared with that of cells transfected with pGenesil-mir-control, Western blot showed a markedly decrease of GLS protein expression (50%) in the cells transfected with pGenesil-mir-218.However, GLS mRNA expression remained unchanged .Conclusion: hsa-mir-218 nega-tively regulates GLS expression at a post-transcriptional level , and this provides an experimental basis for future study of mechanism of GLS expression regulated by mir-218 .