Objective To establish high performance liquid chromatography(HPLC)characteristic chromatograms of Xinqingduyin Granules(composed of Taraxaci Herba,Lonicerae Japonicae Flos,Chrysanthemi Indici Flos,etc.)and content determination of chicory acid and glycyrrhizic acid,and to optimize the preparation process of Xinqingduyin Granules.Methods Using the characteristic chromatograms of Xinqingduyin and the retention rate of chicory acid and glycyrrhizic acid as indexes,we carried out orthogonal experiment to optimize the extraction process of Xinqingduyin,and studied the concentration process.The molding process of Xinqingduyin Granules was conducted by screening the types and dosage of auxiliary materials,then three batches of pilot experiments were carried out.Results HPLC characteristic chromatograms of Xinqingshuyin Granules and the determination methods of chicory acid and glycyrrhizic acid were established.The optimal preparation technology was as follows:8 times amount of water was added,the drug was decocted for 3 times,with 1 hour per time.After the extract was concentrated under reduced pressure at 80℃,the appropriate amount of steviol glycoside and lactose was added into the extract and mixed.One-step granulation and packaging were adopted.The retention rates of chicoric acid and glycyrrhizic acid in the 3 batches of Xinqingduyin Granules,which were prepared on the pilot scale,were(54.56±1.63)%and(54.96±1.08)%,and the rate of finished product was(87.47±0.49)%,respectively.The quality is uniform,and the characteristic map of Xinqingduyin Granules showed high similarity with that of decoction prepared from the same batch of slices.Conclusion The optimized preparation technology is reasonable,feasible and reproducible.This preparation can be used to obtain the granule with similar materials of Xinqingduyin decoction.

Objective: To test the effectiveness of psychological flexibility training on career adaptability among middle school students who undertook psychological courses based on acceptance commitment therapy and the adolescent mental flexibility model(DNA-V), and to provide a reference plan to improve the mental health of middle school students. Methods: This study recruited 110 junior high school students (60 boys and 50 girls) from a middle-school in Beijing. The students were randomly divided by class into a DNA-V face-to-face course group(offline group n=33), a DNA-V online course group(online group n=40), and a regular school psychology course group(control group n=37). Louise Hayes DNA-V intervention program was condensed into a six-hour middle-school DNA-V psychology curriculum. Using the Avoidance and Fusion Questionnaire for Youth and the Career Adaptability Scale, changes in psychological flexibility and career adaptability were measured before(T1), one week after(T2), and two months after (T3) the intervention. Results: Linear mixed models were used for the analysis, while controlling for demographic variables. Psychological flexibility and career adaptability in the offline group were higher at T2 and T3 than at T1(psychological flexibility t=4.22, 3.11; career adaptablity t=3.05, 4.16, P<0.01), while the difference between T2 and T3 was not statistically significant. The psychological flexibility and career adaptability of the online group were not statistically significant at T1, T2, and T3. The psychological flexibility and career adaptability of the control group increased from T1 to T2(t=4.64, 2.47, P<0.05), but T3 decreased back to a level close to T1. Conclusion In terms of both psychological flexibility and career adaptability, the DNA-V face-to-face psychology course resulted in a retention period of at least two months.

Human bioequivalence testing is an important part of evaluating the quality of a formulation. Although these drugs have a large amount of safety data and clinical application data, they may still have ethical risks in healthy subjects. The definition of healthy volunteers, the general inclusion and exclusion criteria, auxiliary inclusion and exclusion criteria, and inclusion and exclusion criteria considering drug specificity are summarized. The basis for determining whether abnormal test values are clinically significant when screening healthy subjects and the considerations for improving the screening pass rate are discussed. It is expected to provide useful reference for the smooth implementation of human bioequivalence testing.

Objective To prepare quality control samples for St.Louis encephalitis virus(SLEV)molecular detection by constructing pseudovirus containing target sequences of SLEV.Methods According to the principles of armored RNA technique, the prM gene sequence of SLEV was cloned into the prokaryotic expression vector to generate recombinant plasmid pSE380-MS2-SLEV.Then, recombinant E.coli transformed with the corresponding plasmid was induced with IPTG to produce recombinant pseudovirus particles.The particles were purified by chloroform and further characterized by double enzyme digestion and transmission electron microscopy.The temperature sensitivity experiments and quantitative RT-PCR were performed to validate the potential of these pseudovirus particles as quality control samples.Results PCR amplification and sequencing analysis confirmed that the prM gene sequence of SLEV was cloned into vector pSE380-MS2.Transmission electron microscopy showed that homogenous spherical particles with a diameter of about 25 nm were produced upon IPTG induction.The SLEV genomic RNA within the pseudovirus particles was resistant to DNaseⅠand RNase A digestion, and remained stable for 20 days at 37℃.These samples were validated with quantitative RT-PCR for SLEV.Conclusion The RNase-resistant and stable pseudovirus particles containing prM fragment of SLEV are constructed successfully, which can be used as positive quality control samples for RNA extraction and molecular detection.

Objective To establish a one-step recombinase polymerase amplification (RPA) method for pathogen screening and rapid detection in the field targeting for five hemorrhagic fever related viruses (Zaire ebola virus, Sudan ebola virus, Marburg virus, Lassa virus and Yellow fever virus). Methods The specific nucleic acid (NA) fragments of each virus were selected as target genes by genome sequence analysis, and the primers and probes for RPA assays were designed according to the sequence. A series of diluted template genes were used for RPA detection to determine the sensitivity. The hemorrhagic fever-related viral nucleic acids were used for RPA detection to determine the specificity. The amplification experiments were carried out at different temperature ranging from 37℃ to 42℃ to validate the reaction temperature range. Results The RPA reaction systems of the five hemorrhagic fever viruses could effectively amplify the target genes, the sensitivities were between 1.5×102 and 1.5×103 copies. No cross reactions existed with the other hemorrhagic fever-related viral genes. Meanwhile, RPA assay could effectively amplify the target genes at 37-42℃. Conclusion The isothermal RPA assays of five hemorrhagic fever viruses are established, which may amply target genes fast and react at a wide temperature range, and be potentially useful for in field pathogens detection.

Until the recent emergence/re-emergence of human-pathogenic viruses in ticks, tick-borne viruses have been neglected as causative agents of human disease (particularly in China). To gain insight into the diversity of tick-borne viruses in Xinjiang Uygur Autonomous Region (northwestern China), we conducted illumina deep sequencing-based screening for virus-derived small RNAs in field-collected Ixodes persulcatus ticks. We found 32, 631 unique virus-matched reads. In particular, 77 reads mapped to the tick-borne group within the genus of Flavivirus, and covered 3.8%-2.4% viral genomes. In addition, 32 unique reads were specific to the Siberian subtype of tick-borne encephalitis viruses (TBEV-Sib) which have never been reported in Chinese TBE loci. We confirmed the potential existence of TBEV-Sib by amplification (using reverse transcription-polymerase chain reaction) of genomic fragments from the envelope gene or 3' genomic terminus from the pools of examined ticks. Both sequences demonstrated high homology to TBEV-Sib strains attached geographically to southern Siberia with nucleotide identity of 97.2%-95.5% and aminoacid identity of 99.4%-98.3%, respectively. In conclusion, we report, for the first time, detection of TBEV-Sib in the natural TBE loci of China. These novel data may provide genetic information for further isolation and epidemiologic investigation of TBEV-Sib.
Animals ; Arachnid Vectors ; virology ; China ; Encephalitis Viruses, Tick-Borne ; classification ; genetics ; isolation & purification ; Encephalitis, Tick-Borne ; transmission ; virology ; Genome, Viral ; Humans ; Ixodes ; virology ; Molecular Sequence Data ; Phylogeny

Objective By prokaryotic expression and purifying the human bocavirus recombinant protein VP2, to establish the indirect enzyme-linked immunoassay for detection of virus. Methods We amplified the human bocavirus recombinant protein VP2 gene fragments from WHL-1 template by PCR , and cloned into the expression vector pET28a, then conversed into the BL21 (DE3) and expressed the fusion protein detected by Western Blot detection , the obtained the antibody and detected the human bocavirus in serum in Guanghzhou area in healthy people. Results The Recombinant prokaryotic expression identified correct by double enzyme, and it could occur specific reaction with the virus positive serum. The best optimal antigen coating concentration were serum multiples and blocking BSA was 2 mg/mL , 1 ∶ 200 and 1%. The best working dilution of enzyme-labeled secondary antibody was 1 ∶ 4 000. The best working hours was 1h. This detection method had good specificity and reproducibility. The cut-off of the indirect ELISA method was 0.1 and the sensitivity and specificity of the developed ELISA method were 92% and 98% respectively. The coincidence rate of determination results by the developed kit and control kit was 97%. Conclusion The competitive ELISA established by prokaryotic expressing and purifying the human bocavirus protein VP2 protein , provides a basis in detecting the human bocavirus serum antibody.

One hundred and seventy six consecutive patients with acute ischemic stroke who received intravenous recombinant tissue plasminogen activator ( rt-PA ) in 4.5 hours from symptom onset during February 2009 to July 2013 were included in the study.Modified Rankin Scale was used to evaluate the recovery of neurological functions.Patients were divided into good ( 0 -1 ) or poor ( 2 -6 ) outcome groups according modified Rankin Scale score.Univariate analysis and multivariate logistic regression analysis were used to determine the differences of clinical data between the two groups.The age of patients with good outcome was significantly lower than that of poor outcome group [ ( 61.4 ±11.5 ) vs.( 69.0 ± 13.2) years,P =0.000].Compared to patients with poor outcomes, patients with good outcome group showed lower rate of diabetes [ 13%( 12/93 ) vs.29%( 24/83 ) , P =0.009 ] , lower blood glucose level [(5.05 ±0.97) vs.(5.83 ±1.72) mmol/L,P=0.020], higher uric acid level[(404.4 ±151.7) vs.(345.6 ±107.5) μmol/L,P=0.028],shorter onset to treatment time [(1.92 ±0.94) vs.(2.30 ±1.01) h, P=0.019],lower baseline National Institute of Health Stroke Scale score [(14.0 ±5.2) vs.(16.0 ± 6.2),P=0.025],lower systolic blood pressure level at 2 h[(140.8 ±18.3) vs.(149.0 ±18.9) mmHg (1 mmHg=0.133 kPa),P=0.005]and 24 h [(137.6 ±21.9) vs.(147.1 ±17.4) mmHg,P=0.009] after thrombolysis.Logistic regression analysis showed that uric acid levels were not related to hemorrhagic transformation independently (P =0.172,OR =0.965,95%CI:0.917 -1.016), but were related to outcome independently (P=0.047,OR=0.957,95%CI:0.916-0.999).

Objective:To investigate the effects of rocuronium in the intravenous anesthesia in children with cleft lip and palate repair.Methods:According to random number table,96 children with cleft lip and palate aged 1-3 years underwent general anesthesia for surgery were divided into 3 groups (n =32).The children in group A,B and C received intravenous injection of rocuronium at 0.6,0.9 and 1.2 mg/kg,respectively.The anesthesia effects were observed.Accelerometry (TOF-Watch(@) SX) was used to monitor neuromuscular transmission.Results:Intubating conditions ranged from good to excellent in all children.The onset time of anesthesia in Group C was significantly shorter(P ＜ 0.05),the duration of neuromuscular block was significantly longer in group C than in group A and B (P ＜ 0.05).The recovery index in group C was significantly longer than that in group A and B (P ＜ 0.05,A vs B,P ＞ 0.05).Muscle relaxant residual incidence in group C was higher than that in group A and B (P ＜ 0.05,A vs B,P ＞ 0.05).Conclusion:Rocuronium at 0.6 mg/kg or 0.9 mg/kg are effective for excellent tracheal intubation,with faster onset time and recovery time,low incidence of postoperative residual curarization(PORC) in children with cleft lip and palate repair.

Objective To discuss the correlation between the level of serum cystatin C and lipids in patients with system lupus erythematosus.Methods Used automatic biochemical analyzer to detect serum cystatin C (CysC),triglyceride (TG),total cholesterol (TC),high density lipoprotein cholesterol (HDL-C),low density lipoprotein cholesterol (LDL-C)and hsCRP levels in 136 cases of SLE patients and 113 cases of healthy people.Data obtained using SPSS13.0 software to carry on sta-tistical analysis.Results Outcome of SLE patients group compared with healthy controls,hsCRP (13.5 ± 4.85 mg/L vs 2.03±0.88 mg/L),CysC (2.63±1.95 mg/L vs 0.85±0.37 mg/L),LDL-C (3.06±1.21 mmol/L vs 2.33±0.41 mmol/L),TC (5.32±2.63 mmol/L vs 4.02±1.67 mmol/L)and TG (1.92±0.83 mmol/L vs 1.44±0.8 mmol/L)were signifi-cantly higher the difference between groups was statistically significant(t=2.45~12.4,P <0.05).Compared with healthy controls,HDL-C (1.12±0.31 mmol/L vs 1.52±0.85 mmol/L)was decreased (P <0.01).In SLE patients group,the ser-um CysC level and hsCRP,TC,TG and LDL-C were positively correlated,and the level of HDL-C was negative to the level of CysC.The health control group was no significant correlation.Conclusion Serum lipid levels of SLE patients were posi-tive to the level of CysC.Suggest that joint detection of SLE patients serum CysC and blood lipids index is helpful to the di-agnosis of SLE treatment and condition monitoring.