Evaluation of performance and application of three nucleic acid extraction methods for quantification of plasma Epstein-Barr virus DNA / 中华检验医学杂志

Objective To evaluate and compare the analytical performances and application values of three nucleic acid extraction methods for quantification of plasma Epstein-Barr Virus ( EBV ) DNA. Methods It used silica membrane spin column , boiling and automated magnetic bead method to extract viral nucleic acid in parallel , and combined real-time fluorescence quantitative PCR assays for quantitative EBV-DNA quantification.The performances of three methods were determined and compared by using the third-party reference materials , and the clinical values were analyzed by pairing detecting 100 NPC patients and 100 healthy subjects in pair .Results The accuracy and imprecision of three methods were all in line with requirements , and the results of clinical samples were linearly correlated . But actually the reproducibility and intermediate imprecision of the magnetic bead method were smaller and stable than those of the spin column method and the boiling method ( all <3％);the limit of detection for the magnetic bead method was 3.334 ×101 IU/ml, better than that of spin column method (4.159 ×101 IU/ml) and boiling method (8.511 ×101 IU/ml);the linear range of the magnetic bead method was 5.4 ×101 -5.4 ×105 IU/ml, slightly wider than that of the boiling method (5.4 ×102 -5.4 ×105 IU/ml); the ability of anti -Hb interference ability of magnetic bead method is better than that of boiling method ;and the positive rate and the mean viral load of the NPC samples measured with the magnetic bead method were significantly higher (95％, 8.342 ×103 IU/ml) than those measured with the spin column method (84％, 4.707 ×103 IU/ml) and the boiling method (78％, 2.571 ×103 IU/ml) ( P all<0.05).Conclusion The automated magnetic bead nucleic acid extraction method offered better analytical performance and higher clinical value for EBV DNA quantification in plasma .

Mild hypothermia protects cerebral ischemia-reperfusion injury in rats by downregulating inhibitor of differentiation 2 / 国际脑血管病杂志

Objective To investigate the protective effect of mild hypothermia on cerebral ischemia-reperfusion injury in rats and the effect of mild hypothermia on the expression of inhibitor of differentiation 2 (Id2) protein.Methods A total of 72 adult male rats were randomly divided into a sham operation group,a normothermia group,and a mild hypothermia group.A model of middle cerebral artery occlusion was induced by a suture method.The mild hypothermia group was treated with low temperature (anal temperature 33±1 ℃,tympanic membrane temperature 31±1 ℃).Modified Neurological Severity Score (mNSS) was used to evaluate neurological deficits,triphenyltetrazolium chloride staining was used to detect infarct volume,and Western blot was used to detect the Id2 expression in the ischemic cortex at ischemia-reperfusion 6,12,24,and 72 h,respectively.ResultsThe mNSS scores in the mild hypothermia group were significantly lower than those in the normothermia group,the infarct volumes were significantly smaller than those in the normothermia group at ischemia-reperfusion 6,12,24,and 72 h (all P<0.001).Western blot analysis showed that the Id2 expressions in the ischemic cortex in the mild hypothermia group were significantly lower than those in the normothermia group at ischemia-reperfusion 6,12,24,and 72 h (all P<0.05).Conclusion s Mild hypothermia can decrease neurological deficits and reduce infarct volume after cerebral ischemia-reperfusion,its mechanism may be associated with the down-regulation of the Id2 expression.

Effect of TanshinoneⅡA on Renal Tumor Growth Factor-beta 1 and Nuclear Factor-kappa B in Diabetic Nephropathy Rats / 广州中医药大学学报

Objective To observe the effect of tanshinone ⅡA on renal transforming growth factor beta 1 (TGF-β1) and nuclear factor-kappa B (NF-κB) p65 mRNA and protein expression in diabetic nephropathy (DN) rats, thus to evaluate the therapeutic effect and mechanism of tanshinone ⅡA. Methods SD rats were used as the experimental animal. DN rat model was induced with 40 mg/kg of streptozocin ( STZ) . The rats were randomized into normal group, model group, and tanshinone ⅡA ( 10 mg·kg -1·d -1, im) group. On the experimental day 30, we examined the body weight, water in-take volume, 24-hour urine protein, fasting glucose ( Glu) , serum creatinine ( Cr) , blood urea nitrogen ( BUN) , total protein ( TP) and albumin ( Alb). Renal slices after periodic acid Schiff staining ( PAS) were used for the observation of renal pathology. Semi-quantitative reverse transcription polymerase chain reaction ( RT-PCR) was used for the detection of renal TGF-β1 and NF-κB p65 mRNA expression, and Western blotting method was used for the measurement of TGF-β1 and NF-κB p65 protein expression in rats of different groups. Results Compared with the normal group, body weight was decreased, water in-take volume and 24-hour urine protein were increased, serum Glu, Cr, and BUN levels were elevated, TP and Alb levels were decreased, renal pathological damage occurred, and renal TGF-β1 and NF-κB p65 mRNA and protein expressin were promoted in the model group (P<0.05 or P<0.01). Tanshinone ⅡA group had an effect on decreasing water in-take volume, 24-hour urine protein, serum levels of Glu, Cr and Bun, increasing TP and Alb levels, relieving renal pathological damage, and reducing the protein and mRNA expression of renal TGF-β1 and NF-κB p65 ( P<0.05 or P<0.01 compared with the model group). Conclusion Tanshinone ⅡA has protective effect on kidney probably through inhibiting renal TGF-β1 and NF-κB p65 expression in DN rats.

Knockout and identification of the surface antigen 43 gene in escherichia Coli JM109 / 重庆医学

Objective To knockout and identify the Antigen 43 (Ag43) in the Escherichia Coli JM109 .Methods Mutation group Ⅱ introns RNA protein complexes (RNP) gene sequence was obtained by Sigma Company′s TargeTron Gene Knockout Sys‐tem and Ag43 gene specific designed PCR primers amplification ,then ,to acquired Ag43 specific recombinant RNP plasmid pACD4K‐Ag4 ,this gene sequence was inserted into the plasmid pACD4K‐C of RNA′s expression .Finally ,pEGFP‐Ag43 was trans‐formed into JM109 and inserted the group Ⅱ intron into the Ag43′s locus by IPTG inducing expression .Results The best insertion locus was between 1 812 and 1 913 .Through the agarose electrophoresis gel ,the RNP gene sequence was consistent with the expec‐ted value (350 bp) .The pEGFP‐Ag43 vector was correctly constructed which was proofed by endonuclease Nhe Ⅰ and Hind ⅡI di‐gestion as predicted products (3 646 and 4 029 bp;7 000 and 550 bp ,respectively ) .The PCR and gene sequence results indicated that the group Ⅱ intron was inserted into the locus between 1 812 and 1 913 in the Ag43 gene .Conclusion Successful knockout of the Ag43 in Escherichia Coli JM109 found basis to further study the Ag43′s function and regard the coli as host bacteria of Ag43 chimeric protein recombinant .

Application of Framework of ICF in Community- based Rehabilitation Assessment for Children with Cerebral Palsy / 中国康复理论与实践

@#Objective To explore the theoretical framework and method of community-based rehabilitation assessment for children with cerebral palsy based International Classification of Functioning, Disability and Health (ICF). Methods The system of assessment of community-based rehabilitation was developed based on the theoretical framework ICF, combined with multi-tools and self-compiled work scales.The system was used for 50 rural children with cerebral palsy after 1-year of community-based rehabilitation. Results and Conclusion The system of assessment is applicable for rural community-based rehabilitation of children with cerebral palsy, with satisfactory efficacy and compliance.

Clear cell sarcoma of gastrointestinal tract：clinicopathologic analyses and review of litera-tures / 临床与实验病理学杂志

Purpose To study the clinicopathologic and genetic features of clear cell sarcoma of the gastrointestinal tract. Methods One case of clear cell sarcoma of the gastrointestinal tract was analyzed by histology, immunohistochemistry, fluorescence in situ hy-bridization ( FISH) , with review of relevant literatures. Results A female patient was admitted to the hospital with a history of 1-week abdominal pain. Computed tomography revealed a mass in right hemicolon. A grey-white ulcerated mass was observed in the colon, in-volving the entire thickness of the colon wall extending into the subserosa. Microscopically, medium-size oval or round cells were ar-ranged in sheets and scattered osteoclast-like multinucleated giant cells were also presented. Immunohistochemically, the tumor cells were strongly positive for S-100 protein, but negative for HMB-45, Melan-A, CD117, CD1a and PCK. FISH showed the rearrangement of EWSR1 in 74% of the neoplasm cells. Conclusions Clear cell sarcoma of the gastrointestinal tract is an extraordinarily rare gastro-intestinal neoplasm that shows peculiar histopathological, immunohistochemical, ultrastructural, and genetic features. Whether clear cell sarcoma-like gastrointestinal tumor represents a distinct entity awaits larger series, including cytogenetic or molecular biological in-vestigation.

Cholinergic dysfunctions are correlated with memory deficits in rats exposed to chronic hypoxia-hypercapnia / 中华神经科杂志

Objective To investigate the relationship between changes of cholinergic system and memory ability impairment in rats long-term exposed to hypoxia-hypercapnia.Methods Forty-eight SD rats were randomly divided into control group,2 weeks hypoxia-hypercapnia group and 4 weeks hypoxiahypercapnia group.The chronic hypoxia-hypercapnia rat model was set up.The memory ability was assessed by eight-arm radial maze.Morphological changes were observed by the HE staining and Nissl staining.Acetylcholine(ACh) content,choline acetyl-transferase (ChAT) activity and acetyl-cholinesterase (AChE) activity in the hippocampus were detected by spectrophotometry,while expression of α7 neuronal nicotinic acetylcholine receptor (α7 nAChR) protein by Western blot.Results Memory ability,especially the working memory was impaired in rats exposed to chronic hypoxia-hypercapnia.And the memory ability decreased more markedly in four weeks group.Compared with those of normoxic rats,the levels of ACh and the activities of AChE and ChAT in the hippocampus of the two weeks group were significantly decreased (ACh:(58.9 ±2.7) vs (47.4 ±3.2) μg/mg (protein) ; AChE:(0.326 ±0.019) vs (0.247 ±0.020) U/mg (protein) ; ChAT:(127.1 ±8.6) vs (90.4 ±6.9) U/g (tissue wet weight),t =7.674,8.139,9.408,all P ＜ 0.05).Compared with the two weeks group,those changes were more obvious in the four weeks group rats (ACh:(47.4 ±3.2) vs (32.5 ±3.2) μg/mg (protein); AChE:(0.247 ±0.020),(0.170±0.019) U/mg (protein); ChAT:(90.4 ±6.9),(55.6 ±6.0) U/g (tissue wet weight),t =9.279,8.036,10.781,all P ＜ 0.05).Compared with the normoxic rats,the expressions of α7 nAChR protein were significantly decreased in two weeks group rats (t =4.481,P ＜ 0.05).Moreover,the expressions of α7 nAChR protein were significantly decreased in four weeks group rats comparing with the two weeks group (t =4.965,P ＜ 0.05).Conclusions An impairment of rat' s memory ability may be induced by hypoxia-hypercapnia,and the injury degree is correlated with the exposure time.Cholinergic system dysfunctions may contribute to the memory function defects in chronic hypoxia-hypercapnia rats.

CT Diagnosis of Endo-bronchial Tuberculosis / 实用放射学杂志

Objective To evaluate CT diagnositic value for endo-bronchial tuberculosis(TB).Methods CT findings of 14 cases with endo-bronchial TB proved by fibro-bronscopy(FOB) were studied retrospectively.Results The bronchiarctia was seen at lobar bronchas or segmental bronchas in all 14 cases.Of them,in 6 cases,the bronchiarctia appeared as conic stenosis accompanied with atelactasis and a large amount of plearal effusions,in 8 cases,the bronchial walls were thichened including centrality stenosis in 3 cases,unilateral stenosis in 5 cases and accompanied with polyus in 3 cases.No hilar mass and opposite "S" sign were found.The "bronchial air sign" was showed at the area of ateectasis.9 cases were accompanied by active pulmonary TB(64.3%).Conclusion CT is of important value in diagnosing endo-bronchial TB and differentiating the tumorous or no-tumorous bronchiarctia,while the diagnonsis at early stage of this disease has to depend on FOB.