To prepare virus-like particles containing FluA/B mRNA as RNA standard and control in Influenza RNA detection, the genes coding the coat protein and maturase of E. coli bacteriophage MS2 were amplified and cloned into D-pET32a vector. Then we inserted 6 histidines to MS2 coat protein by QuikChange Site-Directed Mutagenesis Kit to construct the universal expressing vector D-pET32a-CP-His. In addition, the partial gene fragments of FluA and FluB were cloned to the down-stream of expressing vector. The recombinant plasmid D-pET32a-CP-His-FluA/B was transformed to BL21 with induction by IPTG. The virus-like particles were purified by Ni+ chromatography. The virus-like particles can be detected by RT-PCR, but not PCR. They can be conserved stably for at least 3 months at both 4 degrees C and -20 degrees C. His-tagged virus-like particles are more stable and easier to purification. It can be used as RNA standard and control in Influenza virus RNA detection.
Escherichia coli ; genetics ; metabolism ; Influenza A virus ; genetics ; metabolism ; Influenza B virus ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; RNA, Viral ; genetics ; metabolism ; Recombinant Fusion Proteins ; genetics ; metabolism ; Ribonucleases ; chemistry ; Virion ; genetics ; metabolism

Abstract?AlM:To analyze of the clinical features of acute retinal pigment epitheltis ( ARPE) .?METHODS: The clinical data of 36 ARPE patients ( 40 eyes) attending this center from January 2008 to January 2014 were reviewed retrospectively. Of them, 21 patients (58.3%) were male (male :female=1:0. 71). The mean age was 40. 92±7. 13 years old (range:17~60y). The mean best-corrected visual acuity (BCVA) was 0. 50±0. 26 with a range of 0. 3 ~ 1. 0. Thirty-two patients were unilateral cases. All the patients were examined for BCVA, funds photography, fluorescein fundus angiography ( FFA ) , optical coherence tomography ( OCT) . FFA was shown as three types: type ▏ to multiple black light or grape variety fluorescent spot; Type II for l lesions visible fluorescence leakage; Type Ⅲ lesions with choroid neovascularization ( CNV ) . OCT was the following three forms: multiple RPE lesions layer reflection intermittent, proliferation ( type ▏); pigment epithelial detachment with limitations neural epithelium ( typeII);types l and ll with CNV ( type Ⅲ) .?RESULTS: Ocular fundus showed that the lesions were multiple dark-gray spots with a dark circumscribed area at the macular or nearby in all 40 eyes. FFA showed:21 eyes were type ▏, 17 eyes were type II and 2 eyes were typeⅢ, BCVA between type ▏ and type II was statistically significant (P<0. 05), the same was between type 芋. BCVA between different cases in the same type and between type II, Ⅲ, was no statistical difference ( P>0. 05). OCT showed 21 eyes wwere type ▏, 17 eyes were type II and type Ⅲ 2 eyes. BCVA average between type▏ andIIwas statistically significant (P<0. 05). The mean BCVA was no statistically significant difference between type II and Ⅲ patients (P>0. 05).?CONCLUSlON:ARPE fundus demonstrated the multiple dark gray discrete lesions, the degree of visual impairment related with the presence of pigment epithelial barrier and lesion location. OCT and FFA characterized three types. FFA is shown asblack light orgrape variety fluorescent spot, and is the basis of diagnosis. OCT can display the lesions organization form of each layer clearly. lt plays a more and more important role in the diagnosis and differential diagnosis of ARPE.

ObjectiveTo investigate the correlation between serum C-reactive protein(CRP),tumor necrosis factor-alpha(TNF-α) and atherosclerosis in patients with subclinical hypothyroidism.Methods Seventy-five patients with subclinical hypothyroidism were divided into two groups according to the level of TSH:mild group (42 cases) with TSH between 5.5 and 10.0 mU/L and severe group (33 cases) with TSH ＞10.0 mU/L.Thirty healthy physical examinees or volunteers were selected as control group.TG,LDL-C,HDL-C,TSH,FT4,FT3,CRP,TNF-α and carotid intima-media thickness(CIMT) of all cases were detected and measured.ResultsCIMT of mild,severe and control groups were(0.88 ± 0.20),( 1.12 ±0.21 ),(0.62 ±0.21 ) mm,respectively.CIMT of mild and severe groups were both significantly higher than that of control group,CIMT of severe group was significantly higher than that of mild group (P ＜ 0.01 ).There were no significant differences of CRP and TNF- α among three groups (P ＞ 0.05 ).LDL-C of mild and severe groups were significantly higher than that of control group [ (3.22 ± 0.37),(3.49 ± 0.38 ) mmol/L vs.(2.48 ±0.41 )mmol/L,P ＜0.01 ].HDL-C and TG between mild and control groups had no statistical significance (P ＞0.05).Compared with control and mild groups,HDL-C of severe group decreased [(0.92 ±0.10)mmol/L vs.( 1.21 ± 0.14),( 1.17 ± 0.11 ) mmol/L] and TG increased [ ( 1.50 ± 0.49) mmol/L vs.( 1.11 ±0.53),(1.27 ±0.47) mmol/L](P＜ 0.01 or ＜ 0.05).The levels of TG,LDL-C,TSH,CRP and TNF-α had positive correlation with CIMT (r =0.52,0.37,0.48,0.39,0.45;P ＜0.05 or ＜0.01 ).FT4 was negatively correlated with CIMT (r=-0.24,P＜0.05).HDL-C had no relation with CIMT (r =0.06,P ＞0.05).ConclusionsThere are many abnormalities such as metabolism disorder of blood lipid in patients with subclinical hypothyroidism.The risk of atherosclerosis increases in these patients and inflammatory reaction may not be an important factor of subclinical hypothyroidism complicated with atherosclerosis.

OBJECTIVETo explore the relationship between nitric oxide (NO) in central nervous system and exercise-induced fatigue stress and to study the effect of L-arginine (L-Arg), as a substrate of nitric oxide, on the exercise capacity and NO content in the exhausted rat brain and blood.

METHODSThrough an implanted cannula, the normal saline or L-Arg was microinjected into rat's intracerebroventrical for consecutive four days. Then an acute exhaustive model (on the speed of 18 m/min, an inclination of 5 degrees) was established with animal treadmill. The time of exercise till exhaustion was recorded, and the total workload was calculated that represented the exercise capacity. Nitrate and nitrite (NO3/NO2-, NOx-) levels in blood, hypothalamus and hippocampus were assayed.

RESULTSBoth the time of exercise till exhaustion and total workload in the LArg group increased respectively by 51.8% and 50.08% (P < 0.05), compared with those in the control. The NOx- content in hypothalamus in the L-Arg group (8.93 +/- 1.83) micromol/g pro was larger than that in the control (4.25 +/- 0.79) micromol/g pro, (P < 0.01). There was no significant difference in NOx- content in brain and hippocampus between the two groups. The total workload was positively correlated with NOx- concentration in hypothalamus (P < 0.01). However, there was no correlation between workload and changes in hippocampus NOx- content at fatigue.

CONCLUSIONIntracerebroventricular microinjection of L-Arg may enhance the exercise capacity and lead to up-regulation of NO by means of L-Arg-NO signal path in the hypothalamus. Hypothalamus may be a key site in brain in the modulation of physiological exercise.

Animals ; Arginine ; administration & dosage ; pharmacology ; Hippocampus ; metabolism ; Hypothalamus ; metabolism ; Lateral Ventricles ; Male ; Nitric Oxide ; metabolism ; Physical Conditioning, Animal ; Rats ; Rats, Sprague-Dawley

Through collecting and collating the development process of traditional Chinese medicine dispensing, the development of modern Chinese medicine dispensing on the basis of experience could be promoted. "Heyaofenji", "Hehe", " Heji" in ancient Chinese medicine, herbal medicine literature and law were collected, and then things were sorted out according to traditional Chinese medicine dispensing theory, skills and legal norms. Firstly, "Tang Ye Jing Fa" is the earliest book which marks the rudiment of traditional Chinese medicine dispensing. Secondly, traditional Chinese medicine dispensing theory formed in "Shen Nong's herbal classic". Thirdly, Zhang Zhongjing's "Treatise on Febrile Diseases" marked the formation of Chinese medicine dispensing skills. Lastly, Provisions in Tang Dynasty law marks the development of traditional Chinese medicine dispensing.
Chemistry, Pharmaceutical ; education ; history ; legislation & jurisprudence ; methods ; China ; Dosage Forms ; Drug Compounding ; history ; methods ; Drug and Narcotic Control ; history ; legislation & jurisprudence ; Drugs, Chinese Herbal ; chemistry ; History, Ancient ; Medicine in Literature ; Medicine, Chinese Traditional ; history ; methods

Objective To assess the effectiveness of absorbable screws and metallic devices for treating tibial plateau fractures.Methods Fifty-seven cases of tibial plateau fractures were treated with internal fixation using absorbable screws(Group A),cannulated screws(Group B)and condylar buttress plates(Group C)in our hospital.According to Schatzker classification,there were 15 cases with typeⅠfractures,21 with typeⅡ,nine with typeⅢand 12 with typeⅣ,who were followed up for 6-51 months. Statistical analysis was performed to compare bone union and functional restoration of the knee between three groups.Results Bone union was achieved in all cases.Based on Merchant scale for knee func- tion,excellence rate was 84%(16/19)for Group A,92%(11/12)for Group B and 85%(22/26) for Group C,with no significant statistical difference.Conclusion Absorbable screws is as effective as metallic devices for treating tibial plateau fractures and has advantages of minimal invasion,few complica- tions and avoiding secondary operation.

Objective:To analyze the effects of espO gene knockout on the biological characteristics of enterhemorrhagic Escherichia coli (EHEC). Methods:Two-step methods mediated by the suicide plasmid pCVD442-Δ espO and plasmid pTrc99a were used to construct the espO gene-deleted strain (Δ espO) and the complemented mutant (CΔ espO), respectively. HeLa cells were infected with different EHEC strains to analyze the biological functions and lethal effects of espO gene during infection. Results:PCR, electrophoresis and gene sequencing showed that the Δ espO and CΔ espO mutants were successfully constructed. Compared with the wild-type strain, neither the Δ espO nor CΔ espO mutant showed significant difference in growth rate, indicating that the espO gene had no influence on the growth and replication of EHEC. Furthermore, EspO could activate the tumor necrosis factor receptor (TNF)-induced NF-κB signaling pathway, while the effector protein NleB could inhibit the process. EspO could not inhibit the death of HeLa cells induced by TNF or TNF-related apoptosis-inducing ligand (TRAIL) after EHEC infection. Conclusions:In this study, we successfully constructed the espO gene-deleted and complemented mutants of EHEC and preliminarily analyzed the interaction between espO gene and host cells and the effects of espO gene on cell apoptosis during infection, which provided reference for further research on the in vitro biochemical activity and in vivo pathogenic roles of EspO.

Objective To study the detection rate of early esophageal carcinoma using magnifying endoscopy,and to evaluate the relationship between the imaging patterns of microvasculature change and his- tological diagnosis.Methods Two hundred and fourteen patients with esophageal mucosa roughness,ero- sion,plaque,abnormal color and indentation in conventional endoscopy and 16 healthy volunteers were en- rolled.The magnifying endoscopy images were graded as four patterns by intraepithelial papillary capillary loop(IPCL)changes after iodine dyeing.The biopsies underwent pathologic evaluation.The comparison be- tween the imaging patterns of endoscopy and histological diagnosiswas was evaluated.Results 80.4%(90/ 112)esophagitis was type 2,and 85.7%(12/14)early esophageal carcinoma was type 3 and type 4.The difference was significant between early esophageal carcinoma and normal mueosa(?~2=27.32,P

Bronchiolitis ; diagnosis ; microbiology ; pathology ; Female ; Humans ; Infant ; Leukotriene D4 ; analysis ; Male ; Nasopharynx ; chemistry ; microbiology ; pathology

Objective:To investigate the effect of electroacupuncture (EA) at Ganshu (BL 18) and Shenshu (BL 23) on vascular endothelial growth factor (VEGF) and platelet endothelial cell adhesion molecule-1 (PECAM-1)/CD31 around the cerebral infarction focus in middle cerebral artery occlusion (MCAO) rats and the possible mechanism, thus to provide a new strategy for the treatment of cerebral ischemic stroke by acupuncture. Methods:A total of 180 healthy male Sprague-Dawley (SD) rats were randomly divided into a sham operation group, a model group, an acupoint group and a non-acupoint group, 45 rats in each group. MCAO model was established using the modified line-embolus method in all rats except for those in the sham operation group; rats in the acupoint group were treated with EA at Ganshu (BL 18) and Shenshu (BL 23); rats in the non-acupoint group were treated with EA at the control points; rats in other 2 groups were only subjected to bundling without treatment. Ten rats in each group were randomly selected on the 3rd day, the 14th day and the 21st day after acupuncture stimulation to test the neurological function impairment. The expression levels of CD31 and VEGF were also detected. Results:Compared with the model group and non-acupoint group, the neurological function score of the acupoint group was decreased at each time point, and the differences were statistically significant (P<0.05,P<0.01). The expressions of VEGF and CD31 in each group were the lowest on the 3rd day, reached the peak on the 14th day and still remained at high level on the 21st day. And the differences among groups were statistically significant both on the 14th day and the 21st day (P<0.05,P<0.01). Compared with the model group and the non-acupoint group, the expressions of VEGF and CD31 in the acupoint group were increased, and the differences were statistically significant (allP<0.05). Conclusion: EA at Ganshu (BL 18) and Shenshu (BL 23) can significantly improve the neurological function score of MCAO model rats, and shows protective effect on cerebral ischemia. The protective mechanism may be related to the up-regulation of CD31 and VEGF expression around the cerebral infarction focus in the MCAO model rats and induction of angiogenesis.