Objective To establish a nested-real-time-PCR for the detection of IL-21R mRNA in peripheral blood mononuclear cells(PBMC)of patients with rheumatoid arthritis,and explore its clinical significance.Methods The outer primers of span intron and inner primers were designed for nested-real-time-PCR.To increase specific template,the concentration of primers and enzyme,cycle times were decreased in the first amplification,and the real-time fluorescent quantity assay was performed in the second amplification.The ?-actin gene was used as internal reference,and the results were presented as the ratios of IL-21R mRNA to ?-actin mRNA.Results IL-21R mRNA was detected in 60 patients with RA and 30 healthy controls.The expression of IL-21R mRNA gene in the patients with RA was increased,and the statistical analysis has significant differences(P0.05),but a significant difference was found in the patients of grade III compared with those of grade I and II(P

SOX17,a member of the SOX family of transcription factors,is conserved in many species and plays an important role in the formation of embryo endoderm.A number of studies have found that SOX17 gene expression is silenced or decreased might be due to promoter bypermethylation,which plays a vital role in tumorigenesis and tumor progression,and may contribute to aberrant activation of canonical Wnt signaling pathway in several human malignant tumors.

Lots of studies have identified the relationship between hyperuricemia and cardiovascular diseases. Many factors affect the metabolism of uric acid, such as diet, drug, internal environment of organism, etc. However, the relationship between hyperuricemia and cardiovascular diseases is not quite clear. In this review, we present recently published data about the association between hyperuricemia and selected cardiovascular diseases.

Objective We reported previously that single nucleotide polymorphisms SNP) + 11G ＞ A in intron 3 of the human urate transporter 1 (hURAT1) gene are associated with hyperuncaemia in Han Chinese.The aim of the present study was to evaluate the effect of the variants on hURAT1 function.Methods The wild-type,mutant-type hURAT1 and exon 5-null hURAT1 were constructed,and respectively microinjected into the zebrafish embryo yolks.The subcellular localization of different genotypes of hURAT1 was detected by confocal laser scanning microscope.Results Compared with wild type,the mutant recombinant plasmid transcribed two types of mRNA spliceosome,the wild type and the exon 5-null type.The hURAT1 wild type protein was prominent localized on cell membrane,while the mutant type and exon 5-null hURAT1 proteins were distributed uniform in the cytoplasm but not on the cell membrane.Conclusion The hURAT1 variant + 11 G ＞ A resulted in an alternative splicing of hURAT1 mRNA-exon 5-null type.Its protein product exhibited a different subcellular localization compared with that of wild type.

Objective To investigate the effects of ω-3 polyunsaturated fatty acids (ω-3 PUFA) on inflammatory response of intestine and bacteria translocation in rats with traumatic shock (TS) in order to explore the underlying mechanism.Methods A total of 36 male Wistar rats provided by Academy of Military Medical Sciences Animal Center were assigned randomly (random number) into 3 groups (n =12 in each group):sham operation group,TS model group and PUFA pretreatment group.Rat models of IS were established by comminuted fracture of femur and depletion of blood,and 2 mg/kg ω-3 PUFA or normal saline were injected 12 hours and 2 hours before modeling.Blood specimens were collected and intestinal tissue samples were obtained 120 min after modeling.The serum levels of tumor necrosis factor-o (TNF-α),IL-1β,IL-10 and 8-iso-prostaglandin F 2α (8-iso-PGF2α) were measured with ELISA.Light microscopic examination was carried out for histopathological assessment of the intestina tissue and the intestinal mucosa damage index ( IMDI ) was calculated.The number of marked bacilli found in mesenteric lymph nodes,lung,liver,spleen,and kidney tissues were counted under a fluorescent microscope.The percentages for categorical variables and mean ± SD for continuous variables were expressed. Chi-square test and unpaired t-test were used for comparisons among groups,and statistical significance defined as P ＜ 0.05.Results The levels of TNF-α,IL-1β,IL-10 and 8-iso-PGF2α,the IMDI and the positive rates of bacteria translocation in TS model group were [ (325.14 ±21.17) ng/ml,(26.93 +2.58) μg/L,(7.59 ± 1.26) μg/L,(259.73 +61.32) pg/ml,(4.15 +0.37) and 58.33％,respectively] and those in PUFA group were [ (251.47 + 19.16) ng/ml,(17.81±1.94) μg/L,(9.44±1.85) μg/L,(171.44±39.25) pg/ml,(3.28±0.43) and 36.67％,respectively ].And those biomarkers in both TS group and PUFA group were higher obviously than those in sham group [ (37.02 ±5.54) ng/ml,(2.49 ±0.67) μg/L,(2.93 ±0.74) μg/L,(81.26 ± 15.18) pg/ml,(0.33 ±0.12) and 6.67％,respectively,P＜0.01].Compared with TS model group,the levels of TNF-α,IL-1β and 8-iso-PGF2α,the IMDI and the positive rates of bacteria translocation were lower,and the levels of IL-10 were higher in PUFA group ( P ＜ 0.01 or P ＜ 0.05 ).Conclusions The supplementation of ω-3 PUFA lessens the injury of intestina mucosa after traumatic shock,and it may be associated with the inhibition of inflammatory response by intestine and bacteria translocation.was carried out for histopathological assessment of the intestina tissue and the intestinal mucosa damage index ( IMDI ) was calculated.The number of marked bacilli found in mesenteric lymph nodes,lung,liver,spleen,and kidney tissues were counted under a fluorescent microscope.The percentages for categorical variables and mean ± SD for continuous variables were expressed. Chi-square test and unpaired t-test were used for comparisons among groups,and statistical significance defined as P ＜ 0.05.Results The levels of TNF-α,IL-1β,IL-10 and 8-iso-PGF2α,the IMDI and the positive rates of bacteria translocation in TS model group were [ (325.14 ±21.17) ng/ml,(26.93 +2.58) μg/L,(7.59 ± 1.26) μg/L,(259.73 +61.32) pg/ml,(4.15 +0.37) and 58.33％,respectively] and those in PUFA group were [ (251.47 + 19.16) ng/ml,(17.81±1.94) μg/L,(9.44±1.85) μg/L,(171.44±39.25) pg/ml,(3.28±0.43) and 36.67％,respectively ].And those biomarkers in both TS group and PUFA group were higher obviously than those in sham group [ (37.02 ±5.54) ng/ml,(2.49 ±0.67) μg/L,(2.93 ±0.74) μg/L,(81.26 ± 15.18) pg/ml,(0.33 ±0.12) and 6.67％,respectively,P＜0.01].Compared with TS model group,the levels of TNF-α,IL-1β and 8-iso-PGF2α,the IMDI and the positive rates of bacteria translocation were lower,and the levels of IL-10 were higher in PUFA group ( P ＜ 0.01 or P ＜ 0.05 ).Conclusions The supplementation of ω-3 PUFA lessens the injury of intestina mucosa after traumatic shock,and it may be associated with the inhibition of inflammatory response by intestine and bacteria translocation.

Objective To investigate the effects of ω-3 polyunsaturated fatty acid (ω-3 PUFA) on acute lung injury (ALI) in a rat model of traumatic shock. Methods Thirty-six male Wistar rats aged 3 months were randomly assigned into 3 groups ( n = 12 each): control group (group C) ; traumatic shock group (group TS) and ω-3 PUFA + TS group (group to-3 PUFA) . Traumatic shock was induced by fracture of femur and hemorrhage according to the method described by Feeney in groups TS and ω-3 PUFA. In group ω-3 PUFA, ω-3 PUFA 2 ml/kg was injected via the caudal vein at 12 and 2 h before induction of traumatic shock. Arterial blood samples were taken at 120 min after traumatic shock was successfully induced for determination of serum concentrations of TNF-α, IL-1β, IL-10 and 8-iso-PGF2α by ELISA. The animals were then sacrificed and lungs were removed for-determination of W/D lung weight ratio and microscopic examination. Results Traumatic shock significantly increased serum concentrations of TNF-α, IL-1β, IL-10 and 8-iso-PGF2α, W/D ratio and pathologic scores of lung tissues in groups TS and ω-3 PUFA as compared with group C.ω-3 PUFA significantly attenuated traumatic shuck-induced increase in serum concentrations of TNF-α, IL-1β and 8-iso-PGF2α , W/D ratio and pathologic scores of lung tissues but further increased the serum IL-10 concentration in group ω-3 PUFA as compared with group TS. Conclusion ω-3 PUFA can significantly inhibit the svstemic inflammatory response and ameliorate traumatic shock-induced ALI.

High mobility group box 1 (HMGB 1 ) is a family member of the high mobility group. HMGB1 binds to cell surface specific receptor-receptor for advanced glycation end products (RAGE) and toll-like receptor 2 (TLR2), and thus it exhibits extensive biological activities. HMGB1 participates in pathophysiological processes including inflammatory response and regulation of blood-brain barrier permeability in central nervous system, arid it is closely correlated with the diseases such as ischemia stroke, Alzheimer's disease and gliocytoma.

The study was designed to measure the effect of S.baicalensiswater extract (SBWE) on 3T3-L1 cells and its adiponectin (ADP) mRNA (Adipoq) and promoter luciferase activity.Cell survival rate was determined by MTT assay.The expression of Adipoq was measured by real-time PCR,while the luciferase report systems of Adipoq were used to transfer 3T3-L1 cells.The luciferase activities of the transferred cells were compared by luciferase assay.It was found that the mRNA expression of Adipoq was decreased in comparison with the control group.The luciferase activity showed a stronger ADP promoter activity in 3T3-L1 cells in SBWE treated group than that of control one.In conclusion,SBWE treatment improves the cykomine expression and luciferase reporter gene activity which will be an essential method for further studies of obesity therapy in traditional Chinese medicine.

Objective To explore gene polymorphism of G/A genotype of Fok Ⅰ rs2228570 (G/A) of vitamin D receptor (VDR) gene in Han male population of Chinese coastal area,and thus to investigate the relationship between the gene polymorphism of VDR and gout.Methods Altogether 504 gout patients and 523 healthy controls were enrolled.The possible association between the polymorphism of VDR rs2228570 and gout in Chinese coastal area was investigated and genotype frequencies and allelic frequencies were calculated by realtime PCR with Taqman(R)probe method.Hardy-Weinberg was used to verify the representativeness of the sample.Comparison between the groups were performed withx2 test and t-test.Results The frequencies of GG,AG,and AA genotypes were 32.1％,50.0％,and 17.9％,respectively among gout patients,while they were 27.9％,50.5％,and 21.6％ respectively among the controls.There was no statistically significant difference in VDR rs2228570 genotype frequencies between gout patients and controls(x2 =3.366,P＞0.05).The allele frequencies of G and A in gout cases were different from those in the controls(57.1％,42.9％;53.2％,46.8％;x2 =3.300,P＞0.05).Conclusions Results of the present study suggest that the G/A genotype of VDR Fok Ⅰ rs2228570 of the VDR gene is not associated with gout in male population of Chinese coastal area.

The aim of this study is to develop a rapid and accurately species typing method for Brucella isolates by using High Resolution Melting (HRM ) analysis .Six pairs of primers were used according to the reference for the sequence of pur‐pose gene .Nineteen biotypes of six species Brucella standard strains were identified by PCR‐HRM analysis and this analysis was used to detect the 35 clinical isolates .Results showed Brucella amplified specific melting curves were different from con‐trasted strains with primer Bspp .The six species Brucella standard strains have own characteristic curve shape from each oth‐ers by PCR‐HRM analysis with five pairs of primers .Thirty‐five clinical isolates of Brucella have entirely consistent with PCR‐HRM curve shape with Brucella melitensis standard strains .So ,PCR‐HRM analysis methods can accurately identify Brucella strains ,especially clinical isolated Brucella melitensis ,and may be used in clinical microbiology laboratories .