Venezuelan equine encephalitis (VEE) is a zoonotic disease caused by the Venezuelan equine encephalitis virus (VEEV) complex. This disease has not yet been reported in China, and it is therefore essential to establish a rapid and accurate method for detection of the virus in order to prevent and control this disease. In this study, a one-step real-time quantitative RT-PCR method was developed for the detection of the VEEV complex. A pair of specific primers and a Taqman probe were designed corresponding to a conserved region of the VEEV gene nspl, allowing the detection of all known strains of different sub- types of the virus. Using RNA synthesized by in vitro transcription as template, the sensitivity of this method was measured at 3.27 x 10(2) copies/microL. No signal was generated in response to RNA from Chikungunya virus (CHIKV), nor to RNA encoding the nsp1 fragment of Eastern equine encephalitis virus (EE-EV) or Western equine encephalitis virus (WEEV), all of which belong to the same genus as VEEV. This indicates that the method has excellent specificity. These results show that this one-step real-time quantitative RT-PCR method may provide an effective tool for the detection of VEEV in China.
China ; DNA Primers ; genetics ; Encephalitis Virus, Venezuelan Equine ; classification ; genetics ; isolation & purification ; Encephalomyelitis, Venezuelan Equine ; virology ; Humans ; RNA, Viral ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; methods

Objective To explore the effect on walking ability of the orthotic insole produced by the International College of Biomechanics (ICB) when it is combined with an ankle foot orthosis (AFO).Methods A total of 40 stroke survivors with hemiplegia were randomly divided into an observation group and a control group,each of 20.In addition to conventional medical treatment and rehabilitation,the control group received AFOs and walking rehabilitation training.The observation group accepted walking training with an ICB orthotic insole combined with an AFO.The experiment lasted 4 weeks.Timed up and go test (TUGT) times,10 m maximum walking speed (10 m MWS) and Berg balance scale (BBS) ratings were recorded before and after the treatment.Results Before the treatment there was no significant difference between the group averages in terms of any of the measures.After the treatment,significant improvement was observed in all of the measurements,with the improvements in the observation group significantly better than those in the control group,on average.Conclusion An ICB orthotic insole combined with an AFO results in better improvements in the walking ability of hemiplegic subjects than an AFO alone.

OBJECTIVETo investigate the potential protective effect of the mitochondria-targeted antioxidant Mitoquinone (MitoQ) on post-thaw human sperm.

METHODSSemen samples were collected from 60 normal fertile men, each divided into six parts of equal volume to be incubated at 37 °C in normal saline (G0, control) or in the extender with 2 nmol/L (G1), 20 nmol/L (G2), 200 nmol/L (G3), 2 µmol/L (G4), and 20 µmol/L of MitoQ (G5). After one hour of incubation, the samples were subjected to computer-assisted semen analysis (CASA) for sperm motility, flow cytometry for reactive oxygen species (ROS), thiobarbituric acid assay for the concentration of malondialdehyde (MDA), and MitoTracker fluorescent staining and flow cytometry for the sperm mitochondrial membrane potential (MMP). Then, the semen were cryopreserved with none (B0), 200 nmol/L (B1), and 2 µmol/L of MitoQ (B2), followed by detection of the changes in the ROS, MDA, and MMP of the post-thaw sperm.

RESULTSThe percentage of progressively motile sperm and total rate of sperm motility were significantly higher in G3 ([30.8 ± 10.2]% and [70.6 ± 9.0]%) and G4 ([32.7 ± 13.5]% and [70.3 ± 11.9]%) than in G0 ([17.6 ± 5.0]% and [54.9 ± 11.5]%) (P < 0.05). The level of ROS dropped markedly with the increased concentration of MitoQ, 86.5 ± 31.6 in G3, 93.6 ± 42.0 in G4, and 45.1 ± 15.0 in G5, as compared with 160.8 ± 39.7 in G0 (P < 0.05). The content of MDA was remarkably lower in G3 ([0.9 ± 0.5] µmol/mg) and G4 ([0.9 ± 0.5] µmol/mg) than in G0 ([1.9 ± 1.1] µmol/mg) (P < 0.05), but not in G5 ([1.7 ± 0.7] µmol/mg), which was even higher than in G3 and G4 (P < 0.05). The MMP showed a significant reduction in G5 (1156 ± 216) in comparison with G0 (1701 ± 251) (P < 0.05) but exhibited no remarkable difference between G0 and G1 (1810 ± 298), G2 (1995 ± 437), G3 (1950 ± 334), or G4 (1582 ± 314). The percentage of progressively motile sperm and total rate of sperm motility after freezing-thawing were significantly decreased as compared with those of the fresh semen (P < 0.01), but both were remarkably higher in B1 ([3.2 ± 2.3]% and [ 43.0 ± 9.5]%) than in B0 ([0.8 ± 0.6]% and [26.5 ± 11.4]%) (P < 0.05). The ROS level was significantly lower in B1 and B2 than in B0 (34.6 ± 12. 3 and 37.0 ± 10.5 vs 56.9 ± 14.3, P < 0.05), and so was the MDA content ([1.4 ± 0.5] and [1.4 ± 0.6] µmol/mg vs [2.6 ± 1.0] µmol/mg, P < 0.05), but the MMP was markedly higher in B1 and B2 than in B0 (1010.0 ± 130.5 and 880.6 ± 128.6 vs 721.1 ± 24.8, P < 0.05).

CONCLUSIONAddition of MitoQ to the freezing extender at 200 nmol/L may effectively improve the quality of human sperm and MitoQ is a good protective addictive for human sperm cryopreservation.

Antioxidants ; Cryopreservation ; Humans ; Male ; Malondialdehyde ; analysis ; Membrane Potential, Mitochondrial ; Mitochondria ; Organophosphorus Compounds ; pharmacology ; Oxidative Stress ; drug effects ; Reactive Oxygen Species ; Semen ; Semen Analysis ; Semen Preservation ; Sperm Motility ; Spermatozoa ; drug effects ; Ubiquinone ; analogs & derivatives ; pharmacology

Objective To explore the activity and its possible neural mechanism of brain default mode network by using resting state functional magnetic resonance imaging (fMRI) in patients with mild cognitive impairment (MCI). Methods The 20 amnestic MCI patients and 25 healthy controls were included in this study, and all subjects underwent mini-mental state examination (MMSE), auditory verbal learning test (AVLT) and fMRI. The data were analyzed by amplitude of low frequency fluctuation (ALFF), and the enhanced and weakened regions of ALFF were observed and compared in both MCI patients and healthy controls. Results MMSE and AVLT tests showed that the memory function was seriously impaired in MCI patients compared with healthy controls, which is based on the short and long delayed episodic memory impairment (2.4±1.7 vs. 6.6±1.4, t=3.70, P＜0.01; 2.1±1.6 vs. 6.7±1.5, t=4.16, P＜0.01). The resting state fMRI showed that MCI patients had significant decreases of ALFF in hippocampal formation, parahippocampal cortex and lateral temporal cortex as compared with health controls (t=2.58, 2.43 and 1.75, all P＜0.01), which were closely relevant to the episodic memory. And they had significant increases in temporal-parietal joint and inferior parietal lobule (t=3.14 and 2.77, both P＜0.01). Conclusions MCI patients show significant decreased active intensity of some DMN nodes that is related to episodic memory in resting state. Increased active intensity in MCI patients would be some type of compensation.

Objective To develop a short Chinese version of the center for epidemiological studies depression scale(CES-D).Method A stratified random nationwide sample with 30801 normal people and a mentally ill sample with 415 patients were tested.Items in the short version were chosen according to both the result of independent t-test in current study and the result of reliance and validity.Cut-off scores were offered to check up depressions.Result The internal consistency reliability of the 9-item Chinese version CES-D ranged from 0.85 to 0.88.The test-retest reliability was 0.49(P＜0.001).All item-total correlations were higher than 0.5.The correlation between sumscores of the original scale and sunscores of the Chinese short version ranged from 0.94 to 0.96 (P＜ 0.001) in all the samples.The sum scores of initial diagnosis depression sample were significantly higher than that of treated depression sample and common sample.A two-dimension structure was testified.The cut-off scores of the short version were 10 and 17.Conclusion The 9-item Chinese version of CES-D can be used in various samples with reliable and valid effect.

Objective To investigate the correlation between RDW and disease activity in patients with IBD and evaluate clinical significance of RDW as a potential indicator to monitor IBD activity. Methods 256 patients with IBD were divided into two groups. One was UC group including 136 patients with 80 active period cases and 56 emission period cases. Another was CD group including 120 patients with 75 active period cases and 45 emission period cases. 60 bacillary dysentery diseases and 80 healthy controls were selected as bacillary dysentery group and healthy control group. RDW, Hb, WBC, PLT, CRP, ESR, MCV were tested and monitored with development of disease at different stages. We compared RDW with CRP,ESR, PLT, Hb, MCV parameters. By ROC curve analysis, the sensitivity and specificity of RDW was estimated in identifying the IBD's activity. Results The mean values of RDW in active UC group, remission UC group, bacillary dysentery group and control group were ( 16. 1 ± 2. 7), ( 13.5 ± 2. 1 ), ( 13.0 ± 2. 0)and ( 12. 8 ± 1.8), respectively. There was significant difference among four groups ( F = 51.9, P < 0. 01 ).RDW in active UC group was significant higher than that in remission UC group, bacillary dysentery group and healthy control group ( t = 8. 12, 9. 67, 11.85, P < 0.05) and RDW in remission UC group was significant higher than that in bacillary dysentery group and healthy control group as well ( t = 2. 45, 2. 67,P <0. 05). The mean values of RDW in active CD group, remissive CD group,bacillary dysentery group and control group were ( 16. 9 ± 2. 2 ), ( 13. 8 ± 1.1 ), ( 13.0 ± 2. 0), ( 12. 8 ± 1.8 ). There was significant difference among four groups ( F = 113.9, P < 0. 01 ). RDW in the active CD group was significant higher than that in remission CD group, bacillary dysentery group and healthy control group (t = 11.47,18.63,18. 72, P < 0. 05 ) and RDW in remission CD group was also significant higher than that in bacillary dysentery group and healthy control group ( t = 3.60, 3. 72, P < 0. 05 ). RDW in UC and CD groups demonstrated a positive correlation with CRP and ESR (r=0. 484, 0. 525, 0. 286, 0. 358 and P<0. 01, <0. 01, < 0. 05, < 0. 01, respectively) but an inverse correlation with Hb and MCV (r = -0. 378, -0. 271,- 0. 329, - 0. 298 and P < 0. 01, < 0. 01, < 0. 05, < 0. 01, respectively). In UC groups RDW represented a larger area under ROC curve (0. 8.54) compared with CRP, ESR, PLT, Fib and MCV. When the cut-off value of RDW was 14. 0, the sensitivity and specificity for identifying active UC were 82% (65/80) and 72% (40/56) respectively. In CD groups, the area of RDW under ROC curve was the largest (0. 925 )among all indicators. When the cut-off of RDW was 14.5, the sensitivity and specificity for identifying active CD was 88% (66/75) and 82% (37/45) respectively. Conclusion RDW in patients with IBD is a useful indicator to estimate the IBD activity and predict disease progression.

Objective To investigate Helicobacter pyloric (Hp) infection in children who underwent gastroscopy in Nanjing area.Methods From Jan.2001 to Dec.2010,2 990 pediatric patients with gastrointestinal symptoms who underwent endoscopy and Hp rapid urease test of gastric mucosa specimens.Results There were 2 990 patients who underwent gastroscopy during the 10 years including 1 718 cases of male and 1 272 cases of female.The rate of Hp infection was 53.8％ (924/1 718 cases) and 52.4％ (667/1 272 cases) in male and female,respectively.There was no significant difference (x2 ＝ 0.532,P ＞ 0.05).From Jan.2001 to Dec.2005,611 patients were tested and the rate of Hp infection was 66.9％ (409/611 cases).From Jan.2006 to Dec.2010,2 379 patients were tested and the rate of Hp infection was 49.7％.There was significant difference(x2 =58.13,P =0.01) between the 2 groups.The rate of Hp infection were 49.6％,48.1％,55.2％,60.0％ in 0-6 years group,＞6-9 years group,＞9-12 years group,and ＞ 12 years group,respectively.There was significant difference among the 4 different age groups (x2 =23.66,P ＜ 0.01).The rate of Hp infection in nodular gastritis(61.4％) was higher than that in chronic superficial gastritis (50.6％).There was significant difference between the 2 groups (x2 ＝ 7.42,P ＜ 0.01).The rate of Hp infection in allergic purpura (46.2％) was higher compared with the children with normal gastroscopy results (15.9％).There was significant difference between the 2 groups (x2 =10.19,P ＜ 0.01).Conclusions There is gradually downward trend by year in the rate of Hp infection of children who underwent gastroscopy in Nanjing area by year.The rate of Hp infection is increasing with the age after the age of 6 years.Hp infection may play a role in the nodular gastritis and allergic purpura.

The present study aimed to study the effects of applying mixed probiotics in the feed on growth performance,antioxidant activity and immune function in yellow-feathered broilers.Three hundred sixty yellow-feathered broilers at 10-day-old were divided into 4 groups.Group A was fed with basal diets,group B with basal diets added 50 g/t mixed probiotics,group C with basal diets added 100 g/t mixed probiotics and group D with basal diets added 300 g/t mixed probiotics.Each group included 9 replicates(10 birds per replicates),and the experiment lasted 50 days.Results showed that group D got the highest average daily gain(ADG) of all groups,and compared with the control group A,ADG was increased by 4.11％ (P＜0.05).Group B got the lowest feed-gain ratio of all groups,and the feed-gain ratio was decreased by 4.58％(P＜0.05) compared with the control group A.Group D got significantly increased duodenal villous length,serum antioxidant index and antibody titer(P＜0.05) compared with the control group A.The effects of mixed probiotics on the growth performance,antioxidant activity and immune function were distinct in yellow-feathered broilers,and 100 g/t mixed probiotics was recommended.

Objective To investigate the correlation between serum homocysteine (HCY) and chronic heart failure (CHF) hypercoagulable state in patients.Methods A total of 105 cases of patients with CHF was divided into three groups according to the New York Heart Association (NYHA) classification standard functions:heart functional grade Ⅱ group (42cases),cardiac function grade Ⅲ group (35 cases) and,NYHA class Ⅳ group (28cases).At the same time,40 healthy individuals were regard as the control group.HCY,fibrinogen (Fbg),D-dimer (DDI),HCY,N-terminal pro-brain natriuretic peptide (NT-proBNP) were detected by fasting venous blood samples which were collected within 24 hours after admission.Results Compared to the control group,the expression of Fbg,DDI,HCY and NT-proBNP increased,whereas,antithrombin Ⅲ (AT-Ⅲ) was reduced.Fbg,DDI,HCY,NT-proBNP,and AT-Ⅲ were found in all patient cases.Four groups were compared with each other,except for cardiac function Ⅱ group and the normal group had no significant difference between them (P ＞ 0.05),the difference between both other groups was significantly different (P ＜ 0.05),HCY had a positive correlation with Fbg,DDI,and NT-proBNP (r =0.268,0.295,and 0.404,P ＜ 0.05),and negative correlation with AT-Ⅲ (r =-0.240,P ＜ 0.05).Conclusions HCY might be a reliable indicator as a judge of CHF patients with hypercoagulable state,to detect HCY,FBG,DDI,and AT-Ⅲ in CHF patients.It benefits for judging thrombosis risk and determining the severity of the diseases.Anticoagulant therapy might be beneficial to reduce the long-term adverse events.

Objective To explore the influence of obstruction sleep apnea syndrome (OSAS) on plasma cystatin C (CC) levels in patients with primary hypertension.Methods A total of 244 cases of primary hypertension patients was chosen.The patients were divided into observation group (with OSAS) and control group (without OSAS) according to apnea hypopnea index (AHI).The observation group was then divided into three subgroups:mild OSAS group,moderate OSAS group,and severe OSAS group.The levels of CC were compared.Results First,the plasma CC levels in patients with primary hypertension had no statistical significance in the differences among different grades of hypertension (P ＞ 0.05).Second,CC levels of observation group were significantly higher than control group (P ＜ 0.05).Third,CC levels of the severe group were higher than the moderate group,and the plasma CC levels of the moderate group were also higher than the mild group and control group.Rank correlation analysis and comparison of CC levels and AHI showed that CC levels were positively correlated with AHI (r =0.585,P ＜ 0.01).However,there were no statistically significant differences between CC levels of the mild OSAS group and control group (P ＞ 0.05).Conclusions The patients with OSAS and primary hypertension had higher levels of CC,and aggravated with the progress of the degree of obstruction.CC may be involved in the progression of the disease,a high level of CC may aggravate the condition,it should be early prevention and treatment.