急、慢性布鲁菌病患者核苷酸结合寡聚化结构域样受体蛋白3炎性小体观察

目的比较急、慢性布鲁菌病（简称布病）患者治疗前外周血核苷酸结合寡聚化结构域样受体蛋白3（NLRP3）、半胱氨酸天冬氨酸蛋白酶1（Caspase-1）、白细胞介素1β（IL-1B）和IL-18水平，了解急、慢性布病免疫特点，寻找布病临床治疗免疫靶点。方法将2016年3—4月在乌兰察布市地方病防治中心治疗的急、慢性布病患者（急、慢性组）各42名作为研究对象，同时选取当地健康人群20名作为对照组。布病的诊断标准按《布鲁氏菌病诊断标准》进行。采用酶联免疫吸附试验（ELISA）法测定急、慢性组和对照组血清NLRP3、Caspase-1、IL—1β和IL-18水平。结果急、慢性组和对照组血清NLRP3、Caspase．1、IL-1β和IL．18水平[（1264．40±424．74）、（1350．67±468．93）、（192．96±61．52），（162．74±54．23）、（172．44±60．56）、（120．10±61．52），（47.23±13．79）、（46．68±14．72）、（27．71±8．71），（202．23±65．24）、（169．19±54．33）、（108．62±41．39）ng／L]比较，差异均有统计学意义（F=61．96、6．26、16．68、18．31，P均〈0．01）；与对照组比较，急、慢性组NLRP3、Caspase-1、IL-1β和IL-18水平均明显升高（P均〈0．05）；与慢性组比较，急性组IL．18水平明显升高（P〈0．05），而急、慢性组NLRP3、Caspase-1、IL-1β水平无明显差异（P均〉0．05）。结论急、慢性布病患者通过NLRP3炎性小体发动的固有免疫应答相同．IL-18水平差异可能对急、慢性布病免疫应答产毕了影响．

Cloning and Analysis of a Disease Resistance Gene Homolog from Soybean

Conserved domains e.g. nucleotide binding site (NBS) were found in several cloned plant disease resistance genes. Based on the NBS domain, resistance gene analogs (RGAs) have been isolated previously and were used as probes to screen a soybean (Glycine max L. Merr.) cDNA library. A full-length cDNA, KR3, was obtained by screening the library and rapid amplification of cDNA ends (RACE) method. Sequence analysis revealed that the cDNA is 2 353 bp in length and the open reading frame (ORF) codes for a polypeptide of 636 amino acids with a Toll-Interleukin-1 receptor (TIR) and a NBS domain. Sequence alignment showed that it was similar to N gene of tobacco. The phylogenetic tree analysis of R proteins with NBS from higher plants was performed. The KR3 gene has low copies in soybean genome and its expression was induced by exogenous salicylic acid (SA).

Isolation of Resistance Gene Analogs from Wheat Based on Conserved Domains of Resistance Genes

Two pairs of degenerate primers were designed based on nucleotide-binding site (NBS) and serine/threonine kinase domain. PCR was performed with the primers and cDNA from the Triticum aestivum-Haynaldia villosa translocation line 6VS/6AL. Amplified products were cloned and sequenced. Nine clones with NBS and one with serine/threonine kinase domain were obtained. The NBS clones were classified to six groups according to their nucleotide sequence identities (90% or higher). These resistance gene analogs (RGAs) all have open reading frames (ORF), and their amino acid sequences show high similarity to Yr10 in wheat, Mla1 and Mla6 in barley, RPS2 in Arabidopsis and other resistance (R) genes with conserved motifs. They were preliminarily mapped on the chromosomes of homoeologous groups 1, 2 and 5 of common wheat by nulli-tetrasomic analysis. The 5'-end sequence of an RGA N5 was obtained by 5'-RACE PCR. It encodes six leucine zipper (LZ) and has high sequence similarity to RPS2.

Clinical significance of NOD2/CARD15 and Toll-like receptor 4 gene single nucleotide polymorphisms in inflammatory bowel disease

AIM： To evaluate the role of genetic factors in the pathogenesis of Crohn＇s disease （CD） and ulcerative colitis （UC）, we investigated the single nucleotide polymorphisms （SNPs） of NOD2/CARD15 （R702W, Gg08R and L1007finsC）, and Toll-like receptor 4 （TLR4） genes （D299G and T399I） in a selected inflammatory bowel disease （IBD） population coming from Southern Italy. METHODS： Allele and genotype frequencies of NOD2/ CARD15 （R702W, Gg08R and L1007finsC） and TLR4 （D299G and T399I） SNPs were examined in 133 CD patients, in 45 UC patients, and in 103 healthy controls. A genotype-phenotype correlation was performed. RESULTS： NOD2/CARD15 R702W mutation was significantly more frequent in CD （9.8%） than in controls （2.4%, P = 0.001） and in UC （2.3%, P = 0.03）. No significant difference was found between UC patients and control group （P 〉 0.05）. In CD and UC patients, no significant association with G908R variant was found. L1007finsC SNP showed an association with CD （9.8%） compared with controls （2.9%, P = 0.002） and UC patients （2.3%, P = 0.01）. Moreover, in CD patients, G908R and L1007finsC mutations were significantly associated with different phenotypes compared to CD wild-type patients. No association of IBD with the TLR4 SNPs was found in either cohort （allele frequencies： D299G-controls 3.9%, CD 3.7%, UC 3.4%, P 〉 0.05; T399I-controls 2.9%, CD 3.0%, UC 3.4%, P 〉 0.05）. CONCLUSION： These findings confirm that, in our IBD patients selected from Southern Italy, the NOD2/ CARD15, but not TLR4 SNPs, are associated with increased risk of CD.

Dependence of the E. coli promoter strength and physical parameters upon the nucleotide sequence

The energy of interaction between complementary nucleotides in promoter sequences of E. coli was calculated and visualized. The graphic method for presentation of energy properties of promoter sequences was elaborated on. Data obtained indicated that energy distribution through the length of promoter sequence results in picture with minima at –35, –8 and +7 regions corresponding to areas with elevated AT (adenine-thymine) content. The most important difference from the random sequences area is related to –8. Four promoter groups and their energy properties were revealed. The promoters with minimal and maximal energy of interaction between complementary nucleotides have low strengths, the strongest promoters correspond to promoter clusters characterized by intermediate energy values.

Features of trinucleotide repeat instability in vivo

Unstable repeats are associated with various types of cancer and have been implicated in more than 40 neurode-generative disorders. Trinucleotide repeats are located in non-coding and coding regions of the genome. Studies of bacteria, yeast, mice and man have helped to unravel some features of the mechanism of trinucleotide expansion. Looped DNA structures comprising trinucleotide repeats are processed during replication and/or repair to generate deletions or expansions. Most in vivo data are consistent with a model in which expansion and deletion occur by different mechanisms. In mammals, microsatellite instability is complex and appears to be influenced by genetic, epigenetic and developmental factors.

The mitochondrial Na^＋/Ca^2＋ exchanger may reduce high glucose-induced oxidative stress and nucleotide-binding oligomerization domain receptor 3 inflammasome activation in endothelial cells

Background The mitochondrial Na^＋/Ca^2＋ exchanger, NCLX, plays an important role in the balance between Ca2. influx and efflux across the mitochondrial inner membrane in endothelial ceils. Mitochondrial metabolism is likely to be affected by the activity of NCLX because Ca^2＋ activates several enzymes of the Krebs cycle. It is currently believed that mitochondria are not only centers of energy produc- tion but are also important sites of reactive oxygen species （ROS） generation and nucleotide-binding oligomerization domain receptor 3 （NLRP3） inflammasome activation. Methods ＆ Results This study focused on NCLX function, in rat aortic endothelial cells （RAECs）, induced by glucose. First, we detected an increase in NCLX expression in the endothelia of rats with diabetes mellitus, which was induced by an injection of streptozotocin. Next, colocalization of NCLX expression and mitochondria was detected using confocal analysis. Suppression of NCLX expression, using an siRNA construct （siNCLX）, enhanced mitochondrial Ca^2＋ influx and blocked efflux induced by glucose. Unexpectedly, silencing of NCLX expression induced increased ROS generation and NLRP3 inflammasome activation. Conclusions These findings suggest that NCLX affects glucose-dependent mitochondrial Ca^2＋ signaling, thereby regulating ROS generation and NLRP3 in- flammasome activation in high glucose conditions. In the early stages of high glucose stimulation, NCLX expression increases to compensate in order to self-protect mitochondrial maintenance, stability, and function in endothelial cells.

Dinucleotide microsatellite repeats are essential for the diagnosis of microsatellite instability in colorectal cancer in Asian patients

AIM: The molecular diagnosis of microsatellite instability (MSI) in colorectal cancer (CRC) is based on the analysis of five microsatellite markers. Among them, the two mononucleotide microsatellite repeats are considered more informative for this analysis than the three dinucleotide ones. The aim of this study is to establish the most relevant markers for MSI analysis in colorectal cancers from Asian patients. METHODS: The MSI analysis of 143 CRC cases in a routine molecular diagnostic laboratory was reviewed. Analysis by fluorescence-based PCR of the five recommended microsatellites was performed, followed by data interpretation according to internationally accepted guidelines. The results were analyzed to address (1) the rate of success in the analysis of histopathological samples not specifically preparedfor molecular analysis; (2) the relative importance ofindividual markers in the diagnosis of high-MSI (H-MSI).RESULTS: MSI analysis was unsuccessful in 34 cases (24%), but for tissues archived in recent years the unsuccessful rate was 5%. We found the D2S123 marker the mostvulnerable to inadequate tissue preservation, failing to amplify in 58 instances. Approximately 30% (32/109) of the cases were H-MSI, while 7/109 (6%) were low-MSI.A detailed analysis of the H-MSI cases revealed that the dinucleotide repeats (and D5S346 in particular) were more relevant than the mononucleotide repeats in assigning the correct MSI status. CONCLUSION: The analysis of dinucleotide repeats isessential for the establishment of MSI status in Asian CRC patients.

Nucleotide-binding Oligomerization Domain-1 Ligand Induces Inflammation and Attenuates Glucose Uptake in Human Adipocytes

Objective To investigate the effects of stimulant for nucleotide-binding oligomerization domain 1 （NOD1） on secretion of proinflammatory chemokine/cytokines and insulin-dependent glucose uptake in human differentiated adipocytes. Methods Adipose tissues were obtained from patients undergoing liposuction. Stromal vascular cells were extracted and differentiated into adipocytes. A specific ligand for NOD1, was administered to human adipocytes in culture. Nuclear factor-κB transcriptional activity and proinflammatory chemokine/cytokines production were determined by reporter plasmid assay and enzyme-linked immunosorbent assay, respectively. Insulin-stimulated glucose uptake was measured by 2-deoxy-D-[ 3 H] glucose uptake assay. Furthermore, chemokine/cytokine secretion and glucose uptake in adipocytes transfected with small interfering RNA （siRNA） targeting NOD1 upon stimulation of NOD1 ligand were analyzed. Results Nuclear factor-κB transcriptional activity and monocyte chemoattractant protein-1 （MCP-1）, interleukin （IL）-6, and IL-8 secretion in human adipocytes were markedly increased stimulated with NOD1 ligand （all P〈0.01）. Insulin-induced glucose uptake was decreased upon the activation of NOD1 （P〈0.05）. NOD1 gene silencing by siRNA reduced NOD1 ligand-induced MCP-1, IL-6, and IL-8 release and increased insulin-induced glucose uptake （all P〈0.05）. Conclusion NOD1 activation in adipocytes might be implicated in the onset of insulin resistance.

Coagulation factor Ⅶ gene polymorphisms are not associated with the occurrence or the survival of hepatocellular carcinoma:a report of 37 cases

Objective: Coagulation factor VII(FVII) triggers the extrinsic pathway of blood coagulation. In our previous study, we showed that FVII plays an important role in tumorigenesis of hepatocellular carcinoma(HCC). However, the role of FVII polymorphism in HCC is still unknown. The present study aimed to investigate the relationship between HCC carcinogenesis and single nucleotide polymorphism of FVII.Methods: Thirty-seven HCC patients and 30 healthy donors were recruited in this study. Four common FVII gene polymorphisms– a decanucleotide insertion at position –323(–323 ins10-bp), a G to T substitution at position –401(–401 G/T), a G to A substitution at position –402(–402 G/A), and a T to C substitution at position –122(–122 T/C) – were analyzed by sequencing or commercialized assays using genomic DNA isolated from blood samples. Clinicopathological parameters between control and HCC subjects were compared according to the specific genotypes.Results: The most common nucleotide variation was –402 G/A. However, no statistically significant difference was observed between healthy controls and HCC subjects for all four polymorphisms in terms of genotype distribution and allele frequencies,indicating that these polymorphisms may not affect HCC tumorigenesis. Furthermore, no association was found between–402 G/A polymorphisms and tumor stage, recurrence, and overall survival.Conclusions: Our results indicate that FVII polymorphisms may not be a key factor that clinically impact tumorigenesis and outcomes of HCC, although further investigations should be conducted to confirm our findings.

Saturated Fatty Acid Induces Insulin Resistance Partially Through Nucleotide-binding Oligomerization Domain 1 Signaling Pathway in Adipocytes

Objective To investigate the potential role of nucleotide-binding oligomerization domain 1 （NOD1）, a component of the innate immune system, in mediating lipid-induced insulin resistance in adipocytes. Methods Adipocytes from Toll-like receptor 4 deficiency mice were used for stimulation experiments. The effect of oleate/palmitate mixture on nuclear factor-κB （NF-κB） activation was analyzed by reporter plasmid assay. The release of proinflammatory chemokine/cytokines production was determined by using real-time PCR. Insulin-stimulated glucose uptake was measured by 2-deoxy-D-[SH] glucose uptake assay. Chemokine/cytokine expression and glucose uptake in adipocytes transfected with small interfering RNA （siRNA） targeting NOD 1 upon fatty acids treatment were analyzed. Results Oleate/palmitate mixture activated the NF-κB pathway and induced interleukin-6, tumor necrosis factor-R, and monocyte chemoattractant protein-1 mRNA expressions in adipocytes from mice deficient in Toll-like receptor 4, and these effects were blocked by siRNA targeting NOD1. Furthermore, saturated fatty acids decreased the ability of insulin-stimulated glucose uptake. Importantly, siRNA targeting NOD 1 partially reversed saturated fatty acid-induced suppression of insulin-induced glucose uptake. Conclusion NOD1 might play an important role in saturated fatty acid-induced insulin resistance in adipocytes, suggesting a mechanism by which reduced NOD1 activity confers beneficial effects on insulin action.

Role of Smad7 in inflammatory bowel diseases

Crohn's disease and ulcerative colitis,the major forms of inflammatory bowel diseases(IBD) in man,are complex diseases in which genetic and environmental factors interact to promote an excessive mucosal immune response directed against normal components of the bacterial microflora.There is also evidence that the pathologic process is due to defects in counterregulatory mechanisms,such as those involving the immunosuppressive cytokine transforming growth factor(TGF)-1.Indeed,studies in human IBD tissues and murine models of colitis have documented a disruption of TGF-1 signalling marked by a block in the phosphorylation of Smad3,a signalling molecule associated with the activated TGF-receptor,due to up-regulation of Smad7,an intracellular inhibitor of Smad3 phosphorylation.Knock-down of Smad7 with a specific antisense oligonucleotide restores TGF-1/Smad3 signalling,thus resulting in a marked suppression of inflammatory cytokine production and attenuation of murine colitis.These findings together with the demonstration that Smad7 antisense oligonucleotide is not toxic when administered in mice have paved the way for the development of a Smad7 antisense oligonucleotidebased pharmaceutical compound that is now ready to enter the clinics.In this article we review the available data supporting the pathogenic role of Smad7 in IBD and discuss whether and how Smad7 antisense therapy could help dampen the ongoing inflammation in IBD.

The quantitative expressions of lymphangiogenic factors and metastasis in human colorectal cancers

Objective： The purpose of this paper was to study the expression levels of newly described lymphatic endothelial markers – LYVE-1, Prox-1, podoplanin and 5’-nucleotidase, and their correlation with metastasis of human colorectal cancers. Methods： Tumor and corresponding tumor-side normal tissue samples were obtained from resected specimens immediately after operation. Expression level of each factor was determined by quantitative real-time PCR （RT-PCR） and Western blot technique. Results： Expression levels of lymphatic endothelial markers LYVE-1, Prox-1, podoplanin and 5’-nucleotidase were significantly different in tumor and tumor-side normal groups. Expression levels of Prox-1 and podoplanin were higher in patients with positive lymph node metastasis than those without metastasis. LYVE-1, but not 5’-nucleotidase expression level was higher in both cancer and normal groups. Conclusion： These results indicate that combined quantitative analysis of lymphangiogenic markers LYVE-1, Prox-1 and podoplanin in colorectal cancer specimens may be useful in predicting metastasis of colorectal cancer to regional lymph nodes. However, the role of 5’-nucleotidase in predicting metastasis of colorectal cancer still remains to be further analyzed.

Design of Queue for DNA-based Computer

This paper discusses the design of the queue for DNA-based computer on the point view of data structure. The nucleotide encodings for all components of the queue are given out formally. The linear double-stranded DNA molecules are used as the storage structure of the queue, and the basic bio-operations over the queue are described. Furthermore, the comparison between the queue of the electronic computer and that of DNA-based computer are elucidated. To prove the feasibility of our work, nucleotide encodings for an instance of queue are given out. All the biological technology mentioned in this paper can be practically implemented in the laboratory. Based on this work, other data structures could be developed in DNA-based computer.

Gold nanoparticle-based anodic stripping voltammetry detection of transcription factor NF-kB

Transcription factor and sequence specific DNA interactions play important roles in drug genome and transcription diagnosis. Gold nanoparticles show high sensitivity, stability and compatibility for biological molecules as electrochemical intercalators. Here unimolecular hairpin oligonucleotides were self-assembled onto Au electrode surface and elongation on solid phase was carried out to double strand oligonucleotides with transcription factor NF-r,13 binding site. Gold nanoparticle-catalyzed Ag deposition was detected by anodic stripping voltammetry （ASV） for NF-kB binding. It was indicated that this method for sequence specific DNA binding protein detection shows pronounced specificity, sensitivity and we can find application in transcription regulation research, open reading frame characterization and functional gene inspection by this method.

Effect of electroacupuncture on NF-κB and NLRP3 inflammasome in uterine tissues of rats with primary dysmenorrhea

Objective: To observe the effect of electroacupuncture (EA) on nuclear factor kappa B (NF-κB) and nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) inflammasome in uterine tissues of rats with primary dysmenorrhea (PD), thus to explore the possible mechanism of EA for PD. Methods: Fifty female Sprague-Dawley (SD) rats were randomly divided into a normal group, a model group, an EA at non-acupoint group, an EA at acupoint group and a Western medicine group, with 10 rats in each group. Except for the normal group, rats in the other four groups were treated with estradiol benzoate combined with oxytocin for 11 d to establish PD rat models. From day 1 of the modeling, rats in the normal group and the model group were only properly grasped without any intervention;Guanyuan (CV 4) and Sanyinjiao (SP 6) were selected for EA treatment in the EA at acupoint group;rats in the EA at non-acupoint group were treated with EA at 5 mm away from the acupoints selected above;rats in the Western medicine group were treated with ibuprofen via gavage. Rats in each group were treated for 10-day successively. On the 11th day, except for the normal group, rats in the other groups were intraperitoneally injected with oxytocin (2 U/rat), and the writhing number within 30 min in each group was compared;the pathological changes in rat uteruses were observed by hematoxylin-eosin (HE) staining, and the pathological damage scores were evaluated. Protein expression levels of NF-κB p65, phospho-NF-κB p65, NLRP3, cysteine aspastic acid-specific protease 1 (caspase-1), interleukin (IL)-1β and IL-18 were detected by Western blot. Results: Compared with the normal group, the writhing number increased significantly (P<0.05), and the extensive exfoliation of the endometrium, severe edema, and histopathological score all increased significantly in the model group (P<0.05) as well as the protein levels of NLRP3, caspase-1, IL-1β and IL-18, and the ratio of phospho-NF-κB p65/NF-κB p65 in rat uterine tissues (all P<0.05);compared with the model group, the numbers of writhing reaction decreased within 30 min (P<0.05), the endometrial exfoliation was rare, the edema degree was mild, and the histopathological scores decreased significantly (all P<0.05) in the EA at acupoint group and the Western medicine group;compared with the model group, the phospho-NF-κB p65/NF-κB p65 ratio and the NLRP3, caspase-1, IL-1β and IL-18 protein levels of rat uterine tissues in the EA at acupoint group were significantly lower (P<0.05);compared with the model group, the caspase-1, IL-1β and IL-18 protein levels of the rat uterine tissues decreased significantly (all P<0.05), and the differences in the NLRP3 and phospho-NF-κB p65/NF-κB p65 levels were statistically insignificant (all P>0.05) in the Western medicine group;compared with the Western medicine group, the phospho-NF-κB p65/NF-κB p65 ratio, also the NLRP3, IL-1β and IL-18 protein levels of the uterine tissues decreased significantly in the EA at acupoint group (all P<0.05), while the difference in the caspase-1 level was statistically insignificant (P>0.05);there were no significant differences between the EA at non-acupoint group and the model group in any indicators (all P>0.05). Conclusion: EA at acupoints significantly improves the pain and pathological damages of PD rats. The mechanism may be related to the reduced uterine inflammation via inhibiting NF-κB phosphorylation and NLRP3 activation in uteruses of PD rats.

Regulatory effects of moxibustion on ubiquitin and NLRP3 proteins in colon of ulcerative colitis rats

Objective:To observe the effects of moxibustion on colonic inflammation,and the expressions of ubiquitin and nucleotide-binding oligomerization domain(Nod)-Iike receptor protein 3(NLRP3)proteins in rats with ulcerative colitis(UC),and to explore the anti-inflammatory mechanism of moxibustion in the UC treatment.Methods:Clean grade male Sprague-Dawley(SD)rats were randomly divided into a normal group(NG),a model group(MG),a moxa-stick moxibustion group(MSMG)and a Western medicine group(WMG).UC model was prepared by freely drinking 35 g/L d ext ran sulfate sodium(DSS)solution.Bilateral Tianshu(ST 25)were selected for mild moxibustion treatment in the MSMG;mesalazine solution was intragastrically administrated in the WMG.Rats in the NG and MG were only grasped and fixed as in the MSMG without any treatment.After treatment,hematoxylin-eosin(HE)staining was performed to observe and score the colonic pathological damage under light microscope;immunofluorescence method was used to determine the expression of colonic ubiquitin protein;immunohistochemical method was used to detect the expressions of colonic interleukin(IL)-1β and NLRP3 proteins.Results:The colon tissue was severely injured,and the pathological score was significantly increased in the MG than in the NG(P<0.01),and the protein expressions of ubiquitin,NLRP3 and IL-1β in the colon were significantly increased(all P<0.01).Compared with the MG,the colonic damage was repaired,the inflammation and pathological scores were reduced,and the ubiquitin,NLRP3 and IL-1β protein expressions were decreased in the MSMG and WMG(all P<0.01).Correlation analysis revealed that the ubiquitin protein expression was correlated with the colonic pathological score and the NLRP3 protein expression(r=0.677,P<0.01;r=0.536,P<0.05).Conclusion:Moxibustion can down-regulate the protein expressions of ubiquitin,NLRP3 and IL-1β in the colon of UC rats,which may be one of the mechanisms to promote the repair of colonic inflammatory lesions and exert anti-inflammatory effects.

Synthesis and the anti-HIV activity of novel dinucleotides containing L-isonucleosides

Three dinucleotides containing L-isonucleosides at 5＇-end were synthesized by an elegant phosphoramidite one-pot method. Their binding modes with HIV integrase were simulated and their anti-HIV activities in pseudotyped virus system were examined.

Discussion of the promising effect of electroacupuncture on cognitive improvement in D-galactose-induced aging rats based on NLRP3-ASC-Caspase-1 signaling pathway

Objective:To investigate the effect of electroacupuncture(EA)on cognitive function in D-galactose(D-gal)-induced aging rats,and the correlation between the effect and nucleotide-binding oligomerization domain(NOD)-like receptor protein 3(NLRP3)-ASC-Caspase-1 signaling pathway.Methods:Forty-six male Sprague-Dawley(SD)rats were randomly divided into a control group(n=10),a model group(n=12),an EA-7 d group(n=12)and an EA-21 d group(n=12).Except the control group,the other three groups received 42 consecutive days of intraperitoneal injection of D-gal to establish aging rat models with cognitive dysfunction.The control group received the same amount of normal saline via intraperitoneal injection.Two EA groups were given EA therapy for 21 consecutive days(began from the 22nd day of modeling)or 7 consecutive days(began from the 36th day of modeling)accordingly at Dazhui(GV 14),Baihui(GV 20),Shenshu(BL 23)and Zusanli(ST 36).After modeling/intervention,all four groups received behavioral evaluations by Morris water maze(MWM)test,novel object recognition(NOR)test and step-down passive avoidance(SDPA)test followed by the Western blot(WB)detection of the expression levels of hippocampal NLRP3 inflammasome-associated proteins NLRP3,ASC and Caspase-1.Results:MWM(place navigation test,PNT)results showed that the escape latency in the model group was significantly longer than that in the other three groups(P<0.05),and there was no significant difference among the other three groups on the 1st day of the test(P>0.05).From the 2nd day to the 4th day of the test,there was no significant difference between the EA-21 d group and the control group(P>0.05)in the escape latency;the escape latency was significantly shorter in the EA-21 d group than in the model group and the EA-7 d group(P<0.05).MWM(spatial probe test,SPT)results showed that the time spent in the target quadrant was significantly shorter and platform crossover number was significantly lower in the model group than in the other three groups(P<0.05).The time spent in the target quadrant was longer in the EA-7 d group than in the model group(P<0.05),but was shorter than that in the control group and the EA-21 d group(P<0.05).There was no significant difference in the swimming speed among the four groups(P>0.05).NOR results showed that there was no significant difference in the recognition ratio between the EA-7 d group and the EA-21 d group(P>0.05),and the recognition ratio was significantly higher in the two EA groups than in the model group(P<0.05),but was lower than in the control group(P<0.05).SDPA results showed that the electric shock number was higher in the model group than in the other three groups(P<0.05),and the differences among the other three groups were statistically insignificant(P>0.05).The model group had the shortest step-down latency,followed by the EA-7 d group,the EA-21 d group and the control group in order(P<0.05).The WB results indicated that the expression level of NLRP3 was significantly lower in the control group and the EA-21 d group than in the model group and the EA-7 d group(P<0.05).The expression levels of ASC and Caspase-1 were significantly higher in the model group than in the other three groups(P<0.05),and there was no significant difference among these three groups(P>0.05).Conclusion:NLRP3 inflammasome may be involved in the development of cognitive decline in aging rats;7 consecutive days of EA intervention can partially improve the cognitive impairment in aging rats though the effect is rather limited;21 consecutive days of EA intervention may improve the learning and memory abilities in aging rats via downregulating the expression levels of NLRP3 inflammasome-associated proteins in hippocampus.

A new quantitative structure-retention relationship model for predicting chromatographic retention time of oligonucleotides

An integrated approach is proposed to predict the chromatographic retention time of oligonucleotides based on quantitative structure-retention relationships （QSRR） models. First, the primary base sequences of oligonucleotides are translated into vectors based on scores of generalized base properties （SGBP）, involving physicochemical, quantum chemical, topological, spatial structural properties, etc.; thereafter, the sequence data are transformed into a uniform matrix by auto cross covariance （ACC）. ACC accounts for the interactions between bases at a certain distance apart in an oligonucleotide sequence; hence, this method adequately takes the neighboring effect into account. Then, a genetic algorithm is used to select the variables related to chromatographic retention behavior of oligonuclcotides. Finally, a support vector machine is used to develop QSRR models to predict chromatographic retention behavior. The whole dataset is divided into pairs of training sets and test sets with different proportions; as a result, it has been found that the QSRR models using more than 26 training samples have an appropriate external power, and can accurately represent the relationship between the features of sequences and structures, and the retention times. The results indicate that the SGBP-ACC approach is a useful structural representation method in QSRR of oligonucleotides due to its many advantages such as plentiful structural information, easy manipulation and high characterization competence. Moreover, the method can further be applied to predict chromatographic retention behavior of oligonucleotides.