Objective To explore the changes in inositol requiring enzyme 1 (IRE1),apoptosis signal regulating kinase 1 (ASK1) and c-Jun N-terminal kinase (JNK) mRNA levels in peripheral blood CD4+ T cells of children with acute paraquat (PQ) poisoning.Methods Blood samples of 30 cases of acute PQ poisoning (PQ group),who visited Tianjin Children's Hospital from June 2014 to June 2016,with 18 male and 12 female,aged from 2 to 14 years old,were collected,and the clinical and laboratory data were documented.Peripheral venous blood samples were collected after paraquat was taken.Thirty healthy children at the same age and of the same sex were selected as a healthy control group,18 male and 12 female,aged from 2 to 14 years old.CD4+ T cells in the peripheral blood were separated,and IRE1,ASK1 and JNK mRNA levels in peripheral blood CD4+ T cells were measured by real time polymerase chain reaction (Real-time PCR) method.Specificity of PCR products was validated through agarose gel electrophoresis.The data were statistically analyzed by SPSS 13.0 software.Results All of the 30 children had mucosal lesions,nausea,vomiting and abdomen pain,19 cases with oliguria and anuria,16 cases with alimentary tract bleeding,12 cases with headache and dizziness,11 cases with short of breath,dyspnea and difficult breathing,8 cases with convulsion,5 cases with jaundice.The IRE1,ASK1 and JNK mRNA levels in PQ group were significantly higher than those in healthy control group (1.70 ± 0.16 vs.1.02 ± 0.18,3.56 ± 0.85 vs.1.05 ± 0.31,5.22 ± 0.87 vs.1.01 ± 0.33,t =15.26,15.21,24.78,all P ＜ 0.01).Conclusions PQ increased the expressions of IRE1,ASK1 and JNK in peripheral blood CD4+ T cells,which may be related to PQ-induced oxidative stress and immune activation and lead to a complex cytokine network via endoplasmic reticulum stress and CD4+ T cell apoptosis and then results in the occurrence and development of multiple organ failure.

Objective To shorten the turn around time of positive blood culture results by optimizing the blood culture positive specimen processing flow.Methods In January 26,2015,the microbiology department started the blood culture positive specimen processing flow optimization project,and applied the Lean Six Sigma method in the microbiological process management.The TAT data of 124 positive blood cultures containing Enterobacteriaceae were collected before and after the start of the project in about two months.We analyzed the turnaround time median,mean and standard deviation and reference Z value,process performance index,millions of error opportunities.We decompose the turnaround time into six time periods to find the key points of the process improvement and the influencing factors,and then put forward the reform measures to optimize the blood culture inspection process.MiniTab17.0 statistical software was used to process capability analysis and double sample t test.Results After the implementation of the project,the average turnaround time of the blood culture was shortened from 77.10 h to 64.03 h,improved by 13.06 h(16.94％).Process performance greatly improved in Ppk value increased from 0.49 to 0.88,the benchmark Z value increased from 1.48 to 2.63.After the improvement,except the positive alarm time of blood culture,the mean of the other decomposition time was significantly shorter than before.Conclusions The application of Six Sigma in process management can greatly improve the work efficiency and process performance.This project can save a lot of manpower,material and financial resources,reduce the waiting,shorten turnaround time,that achieve the desired results.

Objective To investigate the effects of conductive education with routine rehabilitation on cerebral palsy. Methods 59 caseswere divided into the conductive group (n=38) and the control group (n=21). The conductive group received conductive education with routinerehabilitation and the control group received routine rehabilitation only. They were assessed with the gross motor function, intelligentand activity of daily living (ADL) before and after treatment. Results The improvement of gross motor was not significantly different betweenthese groups (P>0.05), while the intelligence, ADL improved more in the conductive group than in the control group (P=0.001). ConclusionConductive education can promote the development of the children in ADL and intelligent.

Objective To express recombinant HBHAΔC and HBHAΔN protein,and compare the HBHA series protein activity with each other.It will be provide a experimental basis for the research on clinical diagnostic of HBHA.Methods The HBHAΔC and HBHAΔN gene fragments were cloned and expressed by transforming E.coli BL-21.Test the protein heparin binding ability by CL-6B column.And then added protein to the BCG 7H9 culture medium,to observe the induced BCG aggregation.Results nHB-HA,rHBHA and HBHAΔN protein have heparin binding ability.Meanwhile nHBHA,rHBHA and HBHA Δ C protein have in-duced BCG aggregation effect.Conclusion The HBHAΔC and HBHAΔN protein were successfully obtained.It was proved that the HBHA C-terminal could be combined with heparin and the N-terminal involved could induce the aggregation of BCG.This results provide a basis for further study on molecular mechanism of TB infection and clinical application.

Objective To establish the gas chromatographic(GC) fingerprint of oleic acid of Whitmania pigra Whitman for its quality control. Methods Ten batches of Whitmania pigra from different sources and processed by different methods were analyzed with Agilent 6890N gas chromatography detector on DB-WAX(30 mm × 0.32 mm × 0.25μm)column at the vaporizing temperature of 270℃, column temperature of 130℃and flame ionization detector (FID) temperature of 280℃. We used a software of Similarity Evaluation System for Chromatographic Fingerprint of TCM(Version of 2004A) published by Chinese Pharmacopoeia Commission to calculate GC similarity. Results Oleic acid content of Whitmania pigra processed by different methods had significant differences (F2,7 = 7.350, P = 0.019). The oleic acid content of samples dried after washing with clean water significantly differed from that of the samples processed by alumen or the slices dried naturally(P = 0.021, P= 0.009). The similarity of the fingerprints was in the range of 0.458 - 0.998. The similarity of samples from Lipu of Guangxi Province was the lowest. Conclusion The fingerprints of most of the samples have very high similarity. The established GC fingerprints can be used to effectively identify the qualified or inferior Whitmania pigra products, which will provide some reference for the quality control of Whitmania pigra.

Objective To purify native and recombinant heparin-binding hemagglutinin(HBHA)protein,and investigate the activity of HBHA polyclonal antibody against aggregation of Bacillus CalmetteGuerin(BCG)induced by HBHA.Methods After growing BCG to the stationary phase in the 7H9 liquid medium,the native HBHA protein(nHBHA)was obtained by CL-6B column chromatography.At the same time,the HBHA gene fragment was cloned and expressed by transforming Escherichia coli BL-21.Then the polyclonal antibody against rHBHA was prepared by immunizing rabbit.Different comcentration of the HBHA protein was added to the BCG liquid medium,and the aggregation of the BCG was observed.Then,add the HBHA protein that incubated with anti-HBHA antibodies to the BCG culture medium and observe the aggregation of BCG.Results The purity of native HBHA was 99% and the concentration was 1.016 mg/ml.The expressed product contained 36% of total somtic protein.After purified,the purity of the recombinant HBHA protein was 97.1% and the concentration was 10.98 mg/ml.Both the rHBHA and nHBHA could induce the aggregation of BCG.When then concentration of nHBHA is 0.2μg/ml,BCG could be induced to aggregate,while the rHBHA concentration is 2μg/ml could induce the aggregation.Both aggregations could be suppressed by the polyclonal antibody against rHBHA.Conclusions The native and recombinant HBHA are successfully obtained.It is proved that the rHBHA could induce the aggregation of BCG similar as nHBHA,and polyclonal antibody against rHBHA could also suppress the activity of nHBHA.It suggested that rHBHA could be further used in clinical diagnosis and vaccination.

Advanced schistosomiasis is the most serious clinical type of schistosomiasis. Its diagnosis and treatment are relat?ed to many special departments,such as gastroenterology,general surgery,neurology,endocrinology,radiology,traditional Chinese medicine,blood purification,endoscopy,intervention,and ICU. It is necessary to apply a multidisciplinary treatment (MDT)mode. However,the mode has no universal standard and guide in practice. It is very important for the implementation of MDT mode of advanced schistosomiasis to form a treatment expert team,formulate the formal working procedures,and standard?ize the treatment schedules. The standardized implementation of MDT mode will be important to provide a more effective clinical decision on advanced schistosomiasis.

Objective To verify and monitor the performance of accuracy, precision and comparability of 26 clinical biochemical analytes (29 methods) in the six centers involved in multi-centers reference intervals research, and to ensure the reliability of theirmeasurement results.Methods During the period of the systems evaluating, two levels of commercial quality control materials and fresh frozen human serum reference materials were applied to verify the performance of inter-laboratory precision and accuracy of analysis systems. During the period of samples testing, the commercial quality control materials were measured whenever samples were analysed, the fresh frozen serum reference materials were measured once a month.The coefficient of variations (CVs), bias and total errors were calculated to assess the precision, accuracy and comparability.Results Verification of precision and accuracy: ( 1 ) the ranges of CVs of 29 methods in the six laboratory laboratories were 0.4%-6.0%, the CVs of all 29 methods met the criterion . (2) The overall average bias of the analysis systems of 21 analytes (24 methods) ranged from -5.15%( ALT) to 4.46% ( Ur ) .Among 24 methods the overall average bias of TP, Glu-GOD, Ur, Cl, Ca exceeded the acceptable range.The quality assessment during the period of samples testing:(1) The overall average bias ranged from -1.95%(Ca) to 2.92%(Ur), median 1.26%, they all met the requirements of relevant standards.( 2 ) When commercial control materials were tested, the requirements of CVs were fulfilled for most methods in the six laboratories,and the CVs of TP, Alb, Cl, Ca exceeded the acceptable range.The overall average TE of all methods met the quality specification for the C-N controls material.For the C-P control material, only the overall average TE of TP (5.05%) exceeded thearceptable range while the other methods met the requirement in criterion.Conclusions The performance of precision and accuracy of the analysis systems used in the six laboratories passed the verification.During the period of sample testing, the performance of precision and accuracy of the most methods in the 6 laboratories met the requirements of quality specifications, and the overall performance was good.Because of the limitation of current technology the performance of some methods didn't fulfill the requirement of specifications, and need to be improved.

Objective To investigate the relationship between the phenotypes and the patterns of genetic mutations in the corresponding resistance genes (rpoB, katG, inhA, ahpC, rrs, rpsL, embB and gyrA) in resistant Mycobacterium tuberculosis (MTB) isolates. Methods Rifampicin-resistant gene (rpoB), isoniazid-resistant genes (katG, inhA, ahpC), streptomycin-resistant genes (rrs, rpsL), ethambutol-resistant gene (embB) and quinolinone-resistant gene (gyrA) were amplified by PCR with sequence-specific primers, then mutants screened by single-stranded conformation polymorphism (SSCP) were sequenced. Results rpoB mutation with predominant Ser450Trp pattern was 94. 9% (56/59) in 59 rifampicin-resistant isolates;katG mutation rate was 38. 9% (35/90) and the main pattern was Ser315Thr, but only 3 inhA mutants and no ahpC mutation were determined in 90 isoniazid-resistant isolates;gyrA mutation with main Asp94Gly then Ala90Val pattern was 82.4% (28/34) in 34 quinolinone-resistant isolates;the total mutation rate was 77.4% in 31 streptomycin-resistant isolates, of which 15 isolates mutated in rrs with main pattern A514C or A1041G, 10 isolates mutated in rpsL Lys88Arg;and embB mutation with main Met306Val accounted for 19.4% (6/31) in 31 ethambutol-resistant isolates. Conclusions The results showed that resistance of resistant MTB may be complicated, and DNA sequencing-based mutation analysis could efficiently detect the molecular makers such as rpoB, katG, gyrA, rrs, rpsL and embB in resistant MTB isolates. Meanwhile, it is notable that the rpoB mutation pattern in our isolates is different from previous report, further effort are needed to confirm the characteristics. The spectrum of potential resistance-related mutations in MTB clinical isolates may lay substantial foundation for the rapid molecular diagnosis and rational use of drug to MTB patients.

The medical assistance to advanced schistosomiasis patients established by the Chinese government is a major public facility for patients with advanced schistosomiasis. Since the medical assistance to advance schistosomiasis patients in Hu?nan Province started ten years ago,a set of mature and operable programs with whole program management and related technolo?gies has been developed. The author investigated the data on medical assistance to advanced schistosomiasis patients in Hunan Province during the last 10 years(from 2006 to 2015)retrospectively,and found that the program had high therapeutic effect and high satisfaction degree of both patients and the society. In order to improve the management of the medical assistance to ad?vanced schistosomiasis patients and share our experiences of the whole program management and related technologies with the colleagues of other provinces,this paper mainly illustrates the experiences of the program,as well as the existing problems and related strategies.