Objective To evaluate the efficacy of Yishen-Tongdu decoction combined with sulfasalazine and meloxicamin for ankylosing spondylitis(AS).Methods A total of 86 AS patients were recruited and randomized into an observation group and a control group,43 in each group.The control group was treated by sulphasalazine and meloxicam,while the observation group was treated by Yishen-Tongdu decoction combined with sulfasalazine and meloxicam.The physical function and the disease activity were measured with the Bath Ankylosing Spondylitis Functional Index (BASFI) and the Bath Ankylosing Spondylitis Disease Activity Index (BASDAI),respectively.The erythrocyte sedimentation rate (ESR) and serum C-reactive protein (CRP) were determined.Results The BASFI (3.25 ± 1.18 vs.4.18 ± 0.96;t=4.544,P＜0.01) and BASDAI (2.33 ± 1.46 vs.3.26 ± 1.43;t=5.245,P＜0.01) after treatment in the observation group were significantly decreased than those in the control group.Serum CRP level (8.62 ± 14.71 mg/L vs.12.57 ± 16.32 mg/L;t=3.143,P＜0.05) and ESR (14.93 ± 17.15 mm/h vs.18.61 ± 20.98 mm/h;t=3.615,P＜0.05) in the observation group were significantly lower than those in the control group.The occiput-wall distance (2.07 ± 0.59 vs.2.68 ±0.69 cm;t=5.332,P＜0.01),finger-floor distance (12.88 ± 1.92 vs.13.26 ± 1.71 cm;t=3.593,P＜0.05),and jaw-sternum distance (1.58 ± 0.63 vs.2.43 ± 0.64 cm;t=4.671,P＜0.01) in the observation group were significantly decreased than those in the control group,while the chest expansion (4.99 ± 0.73 cm vs.4.26 ± 0.68 cm;t=4.226,P＜0.01),and Schober test (6.57 ± 0.91 vs.6.13 ± 0.87em;t=3.733,P＜0.01) in the observation group were significantly increased than those in the control group.Conclusion Yishen-Tongdu decoction combined with sulfasalazine and meloxicam can improve the physical function and the disease activity and tts effect is superior to sulfasalazine and meloxicam in patients with AS.

Objective Analysis the bacteria distribution and drug resistance change in our hospital from 2011 to 2013 ,and pro‐vide evidence for clinical rational use of antimicrobial drugs .Methods Isolate all kinds of pathogens from outpatient ,inpatient from 2011 to 2013 in statistics and the analysis of drug resistance .Results The top four highest separation rates were Escherichia coli , Pseudomonas aeruginosa ,Acinetobacter baumannii and Staphylococcus aureus .Methicillin‐resistant staphylococcus aureus (MRSA) separation rate of 3 years were 52 .17% ,57 .9% ,55 .52% respectively .Conclusion Production extended spectrum β‐lactamases (ELBLs) of Escherichia coli has a tendency to increase;the third generation cephalosporins and aztreonam resistance rates also show an upward trend ,Carbapenems Enterobacteriaceae is still the most active drugs;Glycopeptides ,linezolid ,moxifloxacin ,and rif‐ampicin always maintain a high activity against MRSA ;the resistant rates of non‐fermenting bacteria is higher ,we need to concerned about the emergence of multi‐drug resistant .Recommend giving antibiotics under the guidance of susceptibility results .

AIM: Aquaporins(AQP) are very important for the water transport across cell membrane. Aquaporin 7(AQP 7) and aquaporin 8(AQP8) expressed in the rat testis, but the biophysical functions of these two aquporins in testis and the regulatory mechanism of their expression remain unclear. The aim of this study was to find out if the expression of these two aquporins was correlative with the function of testis, and if panaxadiol saponins (PDS) affected their expression. METHODS: 4 weeks, 8 weeks and 12 weeks old rats were divided into saline and PDS injection groups. Semi-quantified RT-PCR method was employed to detect aquaporin 7 and 8 gene expression: using total RNA extracted from rat testis as PCR template, then using optical scanner to measure the OD value of bands on agrose electrophoresis. OD value of target gene products was divided by which of contol gene. The ratio was identified as the quantity of target gene expression. Western blot was also employed to detect expression of these two proteins in the testis. RESULTS: ①OD value of AQP 7 mRNA products in testis of 4 weeks, 8 weeks, 12 weeks rats were 48 227, 51 536, 59 567 respectively and the ratio divided by OD value of control gene was 0.82, 0.85, 0.99; OD value of AQP 8 mRNA products in testis of 4 weeks, 8 weeks, 12 weeks rats were 23 092, 39 302, 43 316 respectively, the ratio divided by OD value of control gene was 0.39, 0.65, 0.72. Thus, AQP 7 and 8 mRNA expression increased with age. ②After PDS injection for 2 weeks. AQP 7 and 8 mRNA expression in 4 weeks old rats increased to 0.97 and 0. 72,which in control group were 0. 82 and 0. 39; the ratio decreased to 0.77 and 0.55 in 8 weeks old rats, which in control group were 0.85 and 0.65. There was no PCR product of neither aquaporins in 12 weeks old PDS injected rats, except products of control gene. ③ Western blot of AQP 7 and 8 protein showed little difference between PDS and saline injected 12 weeks old rats. CONCLUSION: ①Quantity of AQP 7 and AQP 8 expression was related to testis function of rats. ②PDS influenced the expression of AQP 7 and AQP 8 mRNA, perhaps by promoting the secretion of LH in pituitary.

AIM: To find out whether different dosage of rare earth element-lanthanum can influence the expression of aquaporin 7(AQP 7) in the testis of rats. METHODS:Rats were fed with lanthanum nitrate ［La(NO3)3］ and killed 6 months later. Testes were then removed immediately to extract total RNA. Northern blot analysis is performed finally. RESULTS:0.1 mg/kg La(NO3)3 depressed the expression of AQP 7 in rat testis, while 20 mg/kg La(NO3)3 had no significant effect on it. CONCLUSION: AQP 7 expession is found in the rat testis; La(NO3)3 can depress the expression of AQP 7 in the rat testis.

AIM: To find out whether different dosage of rare earth element-lanthanum can influence the expression of aquaporin 7(AQP 7) in the testis of rats. METHODS:Rats were fed with lanthanum nitrate [La(NO 3) 3] and killed 6 months later. Testes were then removed immediately to extract total RNA. Northern blot analysis is performed finally. RESULTS:0.1 mg/kg La(NO 3) 3 depressed the expression of AQP 7 in rat testis, while 20 mg/kg La(NO 3) 3 had no significant effect on it. CONCLUSION: AQP 7 expession is found in the rat testis; La(NO 3) 3 can depress the expression of AQP 7 in the rat testis.

Objective To construct a eukaryotic expression plasmid of pcDNA 3.1-Ag85A-CD226, and to use it as DNA vaccine then further evaluate its immunogenicity through oral administration in a mouse model.Methods The CD226-PCR2.1-ToPo plasmid was used as the template to clone CD 226 gene by PCR.The CD226 gene was then inserted into pcDNA 3.1-Ag85A plasmid to construct the recombinant plas-mid of pcDNA3.1-Ag85A-CD226.After identified by restriction enzyme analysis and sequencing , the re-combinant plasmid was transfected into HEK 293 cells by using lipofection .The expression of Ag85A-CD226 gene in HEK293 cells was detected by RT-PCR, Western blot and indirect immunofluorescence assay .The purified recombinant plasmid was used to prepare the Ag 85A-CD226 DNA vaccine by liposomal encapsula-tion.The vaccine was administered intragastrically to mice .The activities of NK cells , the cytokine levels in the supernatants of spleen cell cultures and the mRNA level of cytokines in the intestines were evaluated to analyze the immunogenicity of Ag85A-CD226 DNA vaccine.Results The Ag85A-CD226 DNA vaccine was prepared successfully .The expression of Ag85A-CD226 fusion protein was detected in HEK293 cells.The activities of NK cells from mice vaccinated with Ag 85A-CD226 DNA vaccine were higher than those from other control groups (P<0.01).The level of TNF-α, IFN-γand IL-2 in the supernatants of spleen cell cul-tures and in the intestines were significantly up-regulated in comparison with other control groups ( P <0.01).The level of IL-4 in the supernatants of spleen cell cultures was down-regulated in the experimental group (P<0.01), but the level of IL-4 in intestines showed no significant difference among the five groups (P>0.05).Conclusion The Ag85A-CD226 DNA vaccine could significantly enhance Th1 type immune responses systemically and in the intestine as in comparison with those vaccinated with single dose of Ag 85A DNA vaccine or CD226 DNA vaccine.

OBJECTIVETo study the membrane mobility of aquaporin 7 (AQP7) by cloning stably transfected CHO cells with expression of pEGFP-C1-AQP7, in which AQP7 cDNA was fused downstream and in frame to pEGFP-C1 gene.

METHODSThe full sequence of AQP7 was amplified by RT-PCR and then recombined in the downstream of the green fluorescent protein gene in the pEGFP-C1 vector. The recombinant vector pEGFP-C1-AQP7 was stably transfected into CHO cells. With fluorescent microscopy, immunocytochemical stain and Western blot, pEGFP-C1-AQP7 showed a predominant intracellular vesicular localization.

RESULTS(1) The sequence of AQP7 cDNA of the Wistar rat was logged into the GenBank (access number: AY157737). (2) Identification demonstrated that pEGFP-C1-AQP7 fusion protein stably expressed in CHO cells. (3) With fluorescence microscopy, pEGFP-C1-AQP7 showed a predominant intracellular vesicular localization.

CONCLUSIONThe CHO cell line with stable pEGFP-C1-AQP7 expression was set up successfully for advanced research.

Animals ; Aquaporins ; biosynthesis ; genetics ; CHO Cells ; Cricetinae ; Cricetulus ; DNA, Complementary ; genetics ; Green Fluorescent Proteins ; biosynthesis ; genetics ; Male ; Molecular Sequence Data ; Rats ; Rats, Wistar ; Recombinant Fusion Proteins ; biosynthesis ; Reverse Transcriptase Polymerase Chain Reaction ; Testis ; metabolism ; Transfection