The Coat protein and Maturase gene of E.coli bacteriophage MS2 was amplified by PCR,then the gene was cloned into pET32a to construct the intermediate vector pET32aCP.The conservative sequence of FMDV internal ribosome entry site(IRES) was cloned into the downstream of pET32aCP bacteriophage gene to construct the prokaryotic expression vector pCPES.The recombinant plasmid pCPES transformed into E.coli strain BL21(DE3) was induced to express with 1mmol/L IPTG.The expression products were purified by sucrose density gradient centrifugation.The expression products observed by TEM were circular viruslike particles,and the diameter of these particles was about 26nm.The stability of viruslike particles was detected,and the viruslike particles was identified by RTPCR.The results showed that the viruslike particles contain the FMDV IRES RNA and have good stability.The viruslike particles have great prospect as the standard and quality control in the area of RNA virus detection.

AIM: Using functional magnetic resonance imaging, to inspect v sual cortex physical reactions stimulated by rotating grating, and to its components. METHODS: On 1.5T MR scanner, GRE-EPI imaging sequence was carried on 9 h discover ealthy volunteers, the visual cortex response data were processed after delinearation by SPM99.RESULTS: Different components of rotating grating excited different areas of the visual cortex. Dramatic response in the central part of the occipital lobe, which was related to white light stimuli, located at primary visual cortex. Response area in bilateral Broadaman 19 areas was related to vision-motion function. And weak response area in the central part of the occipital lobe was related to shape perception.CONCLUSION: Rotating grating conclude plenty of visual information, and it excites different areas of the optic center as a stimuli. fMRI is a valuable equipment to study the physiology of visual cortex.

Objective To explore the main risk factorrelated to the incidence of chroniobstructive pulmonary disease (COPD) among Chinese people in mainland so ato provide the basifothe decision making on COPD prevention .MethodEighpublished literatureof case-control studieon the risk factoof COPD were collected and analyzed quantitatively and synthetically by the metanalysi;the RevMan 5 .2 software waadopted to perform the consistency tesand calculate the pooled oddradio (OR) value and 95% CI .The risk factorwith the Ovalue＞1 were performed the calculation of population attributable risk pro-portion(PARP) .ResultThe pooled oddradio value,95% CI and PARP were smoking OR= 2 .12(1 .58 -2 .86) ,PARP=28 .16% ;occupational exposure OR=1 .82(1 .04-3 .18) ,PARP=11 .60% ;family history of respiratory disease OR=1 .82(1 .36-2 .44) ,PARP=14 .25% ;coal and biomasfuel focooking and heating OR=3 .29(1 .01-10 .67) ,PARP=41 .29% ;low body masindex OR=2 .58(1 .78-3 .74) ,PARP=5 .71% ;low educational degree OR=1 .24(1 .02 -1 .50) ,PARP=12 .93% ;history of re-currenrespiratory tracinfection during childhood OR= 2 .10 (0 .99 -4 .47 ) ,PARP= 13 .39% ;passitive smoking OR= 1 .00 (0 .89-1 .11) .Conclusion Smoking ,occupational exposure ,family history of respiratory disease ,coal and biomasfuel focooking and heating ,low body masindex ,low educational degree and history of recurrenrespiratory tracinfection during childhood are the risk factorinfluencing the incidence of COPD among Chinese people in mainland .

Animals ; Brain Injuries ; blood ; cerebrospinal fluid ; chemically induced ; Endotoxins ; toxicity ; Female ; Male ; Natriuretic Peptide, C-Type ; analysis ; Neurotensin ; analysis ; Rabbits

Objective To investigate the therapeutic effect of quercetin on acute gouty arthritis and its influence in re-nal function in rats. Methods Seventy male Sprague-Dawley rats were randomly divided into normal control group, model group, colchicine group (0.5 mg/kg), allopurinol group (20 mg/kg), quercetin 100, 200 and 400 mg/kg groups (n=10 for each group). Rats were administered various drugs by oral gavage once a day for seven consecutive days throughout the experi-ment. On the fifth day, the animal model of acute arthritis was set up by giving monosodium urate crystal combined with hypo-xanthine. The inflammatory reaction was detected by measuring the circumference of right hind leg anklejoint with a tie line method at 0, 2, 6, 12, 24 and 48 h. The swelling ratio was calculated. The serum levels of uric acid (UA),β2-microglobulin (β2-MG), cystatin C (Cys-C), urea nitrogen (Urea) and creatinine (Cr) were detected by colorimetry and enzyme-linked im-munosorbent assay.Rats were sacrificed at the end of experiment, and the kidney was weighed and the renal index was calcu-lated. Results Treatment with quercetin, colchicine or allopurinol can significantly attenuate swelling rate in rats of acute gouty arthritis. The serum levels of UA were significantly higher in colchicine group and quercetin group than those of nor-mal control group and allopurinol group. The serum levels of UA was significantly lower in allopurinol group than that of nor-mal control group. After 48-h modeling, there was no significant difference in serum UA level between seven groups except allopurinol group. The levels ofβ2-MG and Cys-C were the lowest in normal control group than those of other groups. The se-rum levels of Urea and Cr and renal index were the highest in allopurinol group compared with those of other groups ( P<0.05). Conclusion Quercetin shows a significant effect of anti-inflammatory on acute gouty arthritis in rats. The model es-tablishment may lead to different degrees of renal damage. Quercetin has no protective effect against renal injury, and allopu-rinol aggravates kidney injury.

Objective To detect genetic polymorphism of myeloperoxidase (MPO) gene and catalase (CAT) gene and their activities, and to analyze their relationship with arsenic poisoning caused by coal-burning. Methods One hundred and thirty arsenic poisoning patients were chosen as case group in Jiaole Village, Xingren County, Guizhou Province(an endemic area). One hundred and forty healthy residents living in 13 km away were chosen as control group. Their blood was collected. Polymerase chain reaction-restriction fragment length polymorphism technique(PCR-RFLP) was used to detect polymorphism of MPO-463G/A and CAT-262C/T. Ultraviolet spectmphotometer method was used to detect myeloperoxidase activity. Chromatometry method was used to detect catalase activity. Results The genotype frequency of MPO-463G/A at GG, GA, AA site was 47.24%(60/127), 44.09%(56/127),8.67% (11/127) in case group and 42.34% (58/137),48.17% (66/137)1,9.49% (13/137) in control group, respectively. The difference between the two groups was not significant(χ2 = 0.642, P > 0.05). The genotype frequency of CAT-262C/T, at CC, CT, TT site was 65.60%(82/125),28.80%(36/125),5.60%(7/125) in case group and 76.51%(101/132), 18.94% (25/132) ,4.55% (6/132) in control group, respectively, without significant difference (χ2 =3.845, P>0.05). The relationship between polymorphism of MPO-463G/A and CAT-262C/T and the risk of arsenic poisoning was not found in this study(ORadj= 1.36, 95%CI: 0.74-2.50 for MPO; ORadj=1.35, 95%CI: 0.69-2.63 for CAT). The activities of MPO and CAT were (25.30±8.70)U/L and (2.80± 1.09)×103 U/L in case group, while (22.76±7.59)U/L and (3.90±1.01)×103U/L in control group with a significant difference(F=0.760 for MPO, F=0.855 for CAT, all P < 0.05). The genotype of MPO-463G/A and CAT-262C/T was not found to have relationship with the activities of MPO, CAT(F=1.312,2.822 for MPO; F= 0.151,0.036 for CAT, P>0.05). Conclusions Genetic polymorphism of MPO-463G/A and CAT-262C/T is not found to have relationship with arsenic poisoning. Arsenic can lead to the change of MPO and CAT activity, which, however, may not be affected by MPO-463G/A and CAT-262C/T polymorphism.

CTB protein possessed mucosal adjuvant immunoactivity. The CTB gene was amplified by PCR method from a strain V. cholerae. The nucleotide sequence of CTB gene was 375 bp and shared 96.0%~99.2% homology with other 6 CTB genes. The recombinant plasmid pTWIN1-CTB transformed E. coli strain BL21(DE3) expressed with 0.8 mmol/L IPTG. The molecular weight of expression products was identical with expectative weight by SDS-PAGE electrophoresis. The CTB fusion proteins mainly assembled inclusion bodies and the outputs of proteins were approximately 20% of the total bacterial proteins. The CTB proteins possessed mucosal immunoactivity by GM1-ELISA assay.

OBJECTIVETo observe the therapeutic effect of Xinfeng Capsule (XFC) in treating adjuvant arthritis of rats and its effect on fas, fasL and bcl-2 expression in synovial membrane.

METHODSRats were randomly divided into the normal group, the model group, the model group, the methotrexate (MTX) group, the TPT group and the XFC group. Except for the rats in the normal group, animals were modelled to adjuvant arthritis with Freund's complete adjuvant, and the latter three groups were treated with MTX, Tripterygium wilfordii polysaccharide (TPT) and XFC respectively. The arthritis index (AI), change of body weight (BW) of rats were recorded, and the expressions of fas, fasL and bcl-2 in rats' synovial membrane were determined.

RESULTSCompared with before treatment, the AI in the three treated groups was lowered significantly (P < 0.01, P < 0.05). The BW increment in the XFC group after treatment was insignificantly different to that in the normal group (P > 0.05), while it was significantly lower in the other two treated groups than that in the normal group and the XFC group (P < 0.01). Compared with the model group, fasL expression was increased in the XFC group significantly (P < 0.05), but bcl-2 expression decreased, fas expression showed insignificant difference (P > 0.05). Comparison of the three gene expressions between the three treated groups showed no significant difference (P > 0.05).

CONCLUSIONXFC could decrease AI in rats with adjuvant arthritis in the same way as MTX and TPT, it effected better in increasing BW than the latter two. The effect of XFC might be performed by its action in enhancing fasL expression, inhibiting bcl-2 expression and promoting apoptosis of proliferated synovial membranous cell so as to restrain hyperplasia of synovial membrane.

Animals ; Apoptosis ; Arthritis, Experimental ; drug therapy ; metabolism ; Capsules ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Fas Ligand Protein ; Hyperplasia ; Membrane Glycoproteins ; biosynthesis ; genetics ; Phytotherapy ; Proto-Oncogene Proteins c-bcl-2 ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Wistar ; Synovial Membrane ; metabolism ; pathology ; fas Receptor ; biosynthesis ; genetics

OBJECTIVETo explore effective therapeutic methods for sensorineural hearing loss (SNHL).

METHODSThirty-four cases of mild and moderate SNHL who were definitely diagnosed with pure tone test and by otologists, including 24 cases of insufficiency of the liver and kidney, 10 cases of excess of liver-fire. They were treated with acupuncture at main points, Ermen (TE 21) or Tinggong (SI 19) or Tinghui (GB 2), and Yifeng (TE 17), Shenting (GV 24), Baihui (GB 20), Houding (GV19), Touwei (ST 9), Conger 1-3, Congnao 1-2; and adjuvant acupoints: Huangshu (KI 16), Qihai (CV 6), Guanyuan (CV 4), Taixi (KI 3) were added for insufficiency of the liver and kidney; Zhongwan (CV 12), Tianshu (ST 25), Yanglingquan (GB 34) and Taichong (LR 3) were added for excess of liver-fire. Sequential multipoint pulse stimulation was adopted, twice each week, for 20 consecutive weeks (40 sessions) and then pure tone test was carried out repeatedly.

RESULTSOf 34 cases (58 affected ears), 13 cases were cured and 21 improved, and the various linguistic frequency hearing threshold pure hearing values before and after treatment increased significantly (P < 0.05).

CONCLUSIONAcupuncture and continuous multipoint pulse stimulation has definite therapeutic effect on linguistic frequency hearing loss in the patient of SNHL.

Acupuncture Points ; Acupuncture Therapy ; Hearing Loss, Sensorineural ; Hearing Loss, Sudden ; therapy ; Humans ; Linguistics