This study aimed to investigate the drug susceptibility of wild-type and mutant avian influenza A (H7N9) virus neuraminidase (NA) to oseltamivir and zanamivir. Codon optimized DNA of H7N9 (A/ Hangzhou/1/2013) NA was synthesized and constructed into the pcDNA3.1/His vector (NA(H7N9-WT)). Mutant NA(H7N9-H274Y) and NA(H7N9-R292K) plasmids were constructed by directed mutagenesis PCR using NA(H7N9-WT) plasmid as the template followed by sequencing. NA plasmids were transfected into 293T cells and cell lysates containing NAs were collected 48 h post-transfection. Wild-type and mutant NAs were analyzed by Western blotting and their activities were tested by the 4-MUNANA-based assay. All three NAs were expressed and enzymatic activities were confirmed. The effects of oseltamivir and zanamivir on all three NAs were then tested. It showed that the half maximal inhibitory concentrations (IC50s) of oseltamivir carboxylate on NA(H7N9-WT), NA(H7N9-H274Y) and NA(H7N9-R292K) were 1.6 nM, 15.1 nM, and > 1 000 nM with fold changes of 9 and > 625, respectively. The IC50 values of zanamivir on NA(H7N9-WT), NA(H7N9-H274Y), and NA(H7N9-R292K) were 1.1 nM, 1.4 nM, and 38.0 nM with fold changes of 1.3 and 34, respectively. These results indicated that oseltamivir and zanamivir could significantly inhibit NA(H7N9-WT). NA(H7N9-R292K) showed high-level resistance to both drugs (34-fold and 625-fold) and NA(H7N9-H274Y) was sensitive to both (1.3-fold and 9-fold). These results indicated that both oseltamivir and zanamivir could be used for patients infected with the H7N9 virus. However, when patients carried the H7N9 virus with a NA R292K mutation, other medications would be preferred over oseltamivir or zanamivir.
Antiviral Agents ; pharmacology ; Humans ; Influenza A Virus, H7N9 Subtype ; drug effects ; enzymology ; genetics ; Influenza, Human ; virology ; Microbial Sensitivity Tests ; Mutation ; Neuraminidase ; antagonists & inhibitors ; genetics ; metabolism ; Oseltamivir ; pharmacology ; Viral Proteins ; antagonists & inhibitors ; genetics ; metabolism ; Zanamivir ; pharmacology

Objective To investigate the methods of reducing radiation dose in CT coronary angiography through optimizing individualized scan dosage protocol.Methods Two hundred patients (group A)underwent coronary CTA examination which was performed with fixed 120 kV and variable mA according to their BMI.The mA was set as 150-300 mA(BMI ＜ 18.5 kg/m2),300-500 mA (18.5 kg/m2 ≤ BMI ＜ 25.0 kg/m2),and 500-800 mA(BMI ≥ 25.0 kg/m2).When all examinations were finished,a linear regression was employed to analyze the correlation between mA and BMI,body surface(Suf),image noise(SD)respectively.The results of the analysis were used to formulate a regression equation,which was further used to establish a table list for quick search on how much mA that individualized coronary CTA scan would need.Another 200 patients(group B)enrolled for the individualized scan were scanned under new protocol that previous study established.The tube voltage was 100 and 120 kV.The tube current was variable according to the data in the table list.One-way ANOVA and Kruskal-wallis H test were used for statistics.Results Regression equation between mA and BMI,Suf,SD was:mA =17.984 × BMI + 169.149 × Suf-2.282 × SD-361.039.The SD(group A:32.08 ± 5.80,group B:28.60±4.47),dose index volume(CTDIvol)[group A:(41.97 ± 11.37)mGy,group B:(33.18±10.07)mGy],effective dose(ED)[group A:(10.91 ±3.07)mSy,group B:(8.83 ±2.72)mSv]had significant differences between the two groups(F =43.45,63.71,49.07 respectively,P ＜0.01 for all).The SD and ED results obtained in group B were better than those in group A.Conclusion Better performances were obtained when BMI combined Suf was used as a new individualized protocol than when BMI was used only,which means good image quality and lower radiation dosage in coronary CTA examination.

There have been many studies on the nutrition and the growth status of children from rural and remote western regions of China,whereas researches on children from urban low-income families are scarce.This study aimed to investigate the growth and nutritional status of children under five years of age from urban low-income families in China.There were 169 children aged 25-60 months recruited from Xiangtan and Jilin,two cities with a population of 2.81 million and 4.26 million respectively,in China in this cluster cross-sectional study.Data were collected on demographic and socioeconomic characteristics,the feeding practices and the incidence of anemia and diarrhea.The results showed that the prevalence of low birth weight and macrosomia was 7.l％ and 9.5％ for the two cities,respectively,which was higher than that for other cities in China (1.5％ and 5.9％).Of all the sampled children,14.6％ and 8.2％ suffered anemia and diarrhea,respectively.Multivariate analysis showed that legumes or nuts fed in a 24-h recall increased the risk of anemia (OR=4.9).Children whose caregivers began to introduce complementary foods relatively late would have high diarrhea prevalence (OR=1.4).In conclusion,the prevalence of anemia and diarrhea in under-five children from urban low-income families in China is relatively high.The growth and nutritional status of these children is greatly affected by feeding practices.A series of measures should be taken by relevant government departments to improve the health of these children.

OBJECTIVETo further select the items based on the pre-test version of quality of life scale in patients with viral myocarditis.

METHODSTotally 100 patients with viral myocarditis were enrolled in this study. Methodologies including frequency distribution, discrete trend, t-test, Cronbach's α coefficient, correlation coefficient and factor analysis were applied to select items from different perspectives.

RESULTSA total of 17 items were selected by frequency distribution method from the perspective of central tendency, 15 items were selected by discrete trend method from the perspective of sensitivity, 16 items were selected by t-test method from the perspective of sensitivity and discrimination, 16 items were selected by Cronbach's α coefficient method from the perspective of internal consistency, 12 items were selected by correlation coefficient method from the perspective of representation and independence, and 18 items were selected by factor analysis method from the perspective of representation.

CONCLUSIONItem selection of quality of life scale in patients with viral myocarditis was successfully conducted based on the clinical epidemiological data using a variety of statistical methods.

Adolescent ; Adult ; Female ; Humans ; Male ; Myocarditis ; virology ; Quality of Life ; Surveys and Questionnaires ; Young Adult

Wheat grain peroxidase 1 (WP1) belonged to class III plant peroxidase with cofactor heme, which not only has antifungal activity, but also influences the processing quality of flour. In order to enhance functional expression of WP1 in prokaryotic system by increasing endogenous heme synthesis, we constructed a recombinant plasmid pACYC-A-L containing hemA and hemL of Esherichia coli. Then, we co-transformed it into host strain T7 Express with secretive expression vector (pMAL-p4x-WP1) or non-secretive expression vector (pET21a-MBP-WP1), respectively. The MBP-WP1 fusion protein was further purified by amylose affinity chromatography and its peroxidase activity was assayed using 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonate) (ABTS) as substrate. At 12 h after induction at 28 degree, the extracellular 5-aminolevulinic acid (5-ALA) production of T7 Express/pACYC-A-L was up to 146.73 mg/L, simultaneously the extracellular porphrins also increased dramatically. The peroxidase activity of functional MBP-WP1 obtained from T7 Express/ (pACYC-A-L + pMAL-p4x-WP1) was 14.6-folds of that purified from T7 Express/ pET21a-MBP-WP1. This study not only successfully enhanced functional expression of wheat peroxidase 1 in Esherichia coli, but also provided beneficial references for other important proteins with cofactor heme.
Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; Heme ; biosynthesis ; genetics ; Peroxidases ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Transformation, Genetic

Triterpenes, which have large application potential in the treatment of cancer, are the main active components of genuine medicinal material Alisma orientale (Sam.) Juzep. Farnesyl pyrophosphate synthase (FPPS) is one of the important rate-limiting enzymes in the synthetic pathway of triterpenes. In this study the FPPS full length cDNA of the A. orientale, was cloned via homology-based cloning approach and rapid amplification of cDNA ends (RACE). The full length of the FPPS cDNA was 1 531 bp (accession no. HQ724508), which contained a full 1 032 bp ORF that encoded 343 amino acids. The deduced protein sequence exhibited five conserved motifs, two of which is riched of Asp (DDXXD). The result of real-time quantitative PCR (QRT-PCR) showed that FPPS gene was expressed in different organs of A. orientale. The expression increased from October to the first ten-day period of December, and then decreased. The FPPS gene expression was higher in leaves but lower in leafstalk, tuber and root. HPLC analysis of active components 23-acetyl-alismol B of A. orientale. during different periods indicated that its change trend should be consistent with FPPS gene expression. It can be primarily deduced that FPPS gene should be an important control point in the synthetic pathway of Alisma terpenes. This study may facilitate the quality of medicinal plants through gene engineering in the future.
Alisma ; enzymology ; genetics ; Amino Acid Sequence ; Base Sequence ; Cloning, Molecular ; Computational Biology ; Conserved Sequence ; DNA, Complementary ; genetics ; DNA, Plant ; genetics ; Gene Amplification ; Geranyltranstransferase ; genetics ; isolation & purification ; metabolism ; Molecular Sequence Data ; Phylogeny ; Plant Leaves ; enzymology ; genetics ; Plant Roots ; enzymology ; genetics ; Plants, Medicinal ; enzymology ; genetics ; RNA, Messenger ; metabolism ; Real-Time Polymerase Chain Reaction ; methods

OBJECTIVETo investigate the effect of Compound Qingqin Liquid (CQL) on the expression level of angiotensin II (Ang II) and COX-2 mRNA transcription and protein expression in the renal tissue of rats with uric acid nephropathy.

METHODSSD rats were randomly divided into the blank control group, the model group, the positive drug group, the high, moderate, and low dose CQL group according to number randomization principle. The model was established by gastrogavage of adenine, accompanied with yeast feeding. Distilled water was given by gastrogavage to rats in the blank control group and the model group. Allopurinol at the daily dose of 9.33 mg/kg was given by gastrogavage to rats of the positive control group. CQL at the daily dose of 3.77 g/kg, 1.89 g/kg, and 0.09 g/kg was respectively given by gastrogavage to rats in the high, moderate, and low dose CQL groups. All treatment lasted for 6 weeks. Rats were randomly divided at week 4 (3 in the blank control group, and 6 in the rest groups), and the rest rats were killed at week 6. The renal tissue was extracted. The expression level of Ang II and COX-2 mRNA transcription were detected by RT-PCR. The expression level of Ang II was detected by ELISA. The expression level of COX-2 protein was detected by Western blot and immunohistochemical assay.

RESULTSCompared with the blank control group, except the mRNA expression of Ang II at week 4, the mRNA and protein expression of Ang II and COX-2 obviously increased at week 4 and 6 in the model group (P < 0.01, P < 0.05). The COX-2 protein expression at week 4 was obviously lower in the high and moderate dose CQL groups than in the model group and the low dose CQL group (P < 0.05); the average integral of optical density value was obviously lower in the positive control group than in the model group. Except the mRNA expression of Ang II in the high dose CQL group at week 6, the mRNA and protein expression of Ang II obviously decreased in the positive control group and each dose CQL group (P < 0.01, P < 0.05). Of them, the effects were better in the high and moderate dose CQL groups than in the positive control group and the low dose CQL group (P < 0.05, P < 0.01). Besides, the mRNA expression of COX-2, the average integral of optical density value were obviously lower in the positive control group and each dose CQL group than in the model group (P < 0.05). The protein expression of COX-2 was obviously lower in the high and moderate dose CQL groups than in the model group (P < 0.05). Of them, the mRNA expression of COX-2 was better in the moderate dose CQL group than in the positive control group (P < 0.05); the protein expression of COX-2 was better in the high dose CQL group than in the low dose CQL group (P < 0.05).

CONCLUSIONCQL was capable of lowering the expression level of Ang II, COX-2 mRNA transcription and protein expression, thus suppressing the inflammatory pathological injury of the renal tissue.

Angiotensin II ; metabolism ; Animals ; Cyclooxygenase 2 ; genetics ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Kidney ; metabolism ; Kidney Diseases ; drug therapy ; metabolism ; Male ; RNA, Messenger ; genetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Uric Acid

OBJECTIVETo investigate the effect of compound qingqin liquid (CQL) on Toll-like receptor 2 (TLR2) and toll-like receptor 4 (TLR4) in rats with urate nephropathy, and to explore its renal protection mechanism.

METHODSTotally 55 SD rats were randomly divided into 5 groups, i.e., the normal control group (n =5), the model group (n =10), the positive drug group (n=10), and the high-, medium-, low-dose CQL groups (n=10) respectively. The urate nephropathy model was induced by intragastrically administering adenine and feeding yeast. Distilled water was intragastrically administered at the daily dose of 10 mL/kg to rats in the normal control group and the model group. Allopurinol was intragastrically administered at the daily dose of 9.33 mg/kg to rats in the positive control group. CQL was intragastrically administered at the daily dose of 3.77, 1.89, 0.94 g/kg to rats in the high-, medium-, and low-dose CQL groups. Rats of each group were executed in batches at the 4th and 6th week respectively. Their kidney tissues were taken out to determine the mRNA transcription level of TLR2 and TLR4 by reverse transcription-polymerase chain reaction (RT-PCR). The protein expression level of TLR2 and TLR4 were determined by Western blot. The protein expression level of TLR4 was also detected by immunohistochemical assay.

RESULTSAt week 4 and 6, the protein expression of TLR2 and TLR4 as well as the mRNA transcription of TLR4 increased in the model group, when compared with the control group (P < 0.05, P < 0.01). Compared with the model group, there was no statistical difference in the transcription level of TLR2 mRNA or TLR4 mRNA among the 3 CQL groups (P > 0.05) at week 4 and 6. Additionally, at week 6, the protein expression of TLR4 and TLR2 could be reduced by CQL (P < 0.05, P < 0.01).

CONCLUSIONCQL might protect kidney tissue against inflammatory injury by inhibiting the protein expression levels of TLR2 and TLR4.

Animals ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Kidney ; drug effects ; metabolism ; Kidney Diseases ; drug therapy ; metabolism ; Male ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley ; Toll-Like Receptor 2 ; genetics ; metabolism ; Toll-Like Receptor 4 ; genetics ; metabolism ; Uric Acid

Translational medicine research, as a bridge of basic medicine and clinical medicine, has become the foci of medicine and developed rapidly. To study the research hotspots and frontiers of translational medicine in re-cent 20 years,Web of Science data sources,bibliometrics methods and visualization techniques were used to study the translational medicine from the aspects of time and space and subjects, which may support the translational medicine research of China.

Objective　To investigate the surgical management of a gun shot wound of blood vessels and immersied, and evaluate its primary effect. Methods　100 rabbits were divided randomly into simple wounded group(SWG,n=50) and seawater immersion group(SIG,n=50).F emoral arteries were impacted by 0.38 gram steel spheres with velocity of 600 ～800 meters per second fired by 7.62 mm rifle. Animals in SIG were immersed in artificial seawater (pH 8.2～8.4, salinity 25.4,temperature 21℃) for 60 min, o f which those in SWG were spared. Grossly injuried artery was excised and restor ation of blood flow was reconstructed by end-to-end anastomosis or reversed au togenous venous grafting or cryopreserved arterial allografting. At 24 h,7,1 4,21 days after operation, blood flow was examined by Doppler ultrasonic detecti on and part of anastomotic sites and graft were collected for pathological obser vation. Results　In completely transected injury, the patency in SIG was 80.00%,while that in SWG was 86.67% in the first 3 weeks. In arterial c ontused injury ,patency in SIG was 86.67%,and that in SWG was 82.35% at the same time. Thrombosis occurred mostly in the first postoperative week. Atypical endo thelial cells were found at the anastomosis sites in the first postoperative week, and the anastomosis sites were lined with endothelium in 3 weeks postopera tively. Conclusion　Early curative effect could be obtained. Whe n grossly injuried artery is excised and followed by a routine surgical procedur e in the treating gunshot wounds immersed in seawater.