Rapid Determination of 13 Sedative Substances and Their Metabolites in Blood by LC-MS/MS / 法医学杂志

Objective To establish an analysis method for simultaneous determination of 13 sedative substances and their metabolites in blood by liquid-liquid extraction and liquid chromatography-tandem mass spectrometry （LC-MS/MS） technology and to apply the method to actual cases. Methods The samples were extracted with ethyl acetate after an internal standard was added. The extract was condensed until it was nearly dry and then its residues were dissolved with methanol, filtered through 0.22 μm filter and finally determined. The 13 sedative substances and their metabolites were separated through the C18 chromatographic column, then gradient elution was performed on them with methanol and 20 mmol/L ammonium formate （containing 0.1% formic acid） solution. After that, they were determined in the electrospray positive ion mode and quantified by internal standard method. Results The 13 sedative substances and their metabolites in blood showed good linearity in the range of 5-200 μg/L with correlation coefficients ranging from 0.990 3 to 0.999 8. The detection limits were 0.1-1.0 μg/L. Recovery rates of sedative substances were in the range of 71.2%-93.4% when solutions with concentrations of 10, 50 and 200 μg/L were added. The deviations of intra-day and inter-day relative standard deviations （RSD） were not more than 8.6%. Accuracies （bias） were within ±9.8%. Conclusion This method is rapid, simple, effective and sensitive, and can be applied to analysis of 13 sedative substances and their metabolites in blood in forensic toxicology.

Establishment and application of multiplex direct PCR for rapid detection of common food borne pathogens in swine products / 中国人兽共患病学报

We established a multiplex direct PCR for rapid detection of E.coli,Salmonella,Staphylococcus aureus,Listeria and Yersinia enterocolitica bacteria.Multiplex direct PCR primers were designed according to gene sequences of phoA (E.coli),inv A (Salmonella),nuc (S.aureus),hl y (Listeria),and ail (Y.enterocolitica).After the multiplex direct PCR were established,the specificity and sensitivity of primers were detected.Then,multiplex direct PCR was applied to examine 60 swine product samples,the detection specificity,accuracy and positive predictive value were calculated compared with the gold standard culture method.Results showed that multiplex direct PCR primers could be used for specific detection of E.coli,Salmonella,S.aureus,Listeria and Y.enterocolitica,with the minimal detectable limit of 10,1,100,1 and 1 CFU,respectively.For the examination of 60 swine product samples using multiplex direct PCR,15 were positive for E.coli,6 positive for Salmonella,21 positive for S.aureus,20 positive for Listeria,and 35 positive for Y.enterocolitica,with all positive detection rates higher than that of culture.The total detection sensitivity was 100％,accuracy was 94％,and positive predictive value was 81.44％.Multiplex direct PCR could be used for specific and sensitive detection of common food-borne pathogens,and the testing time was shorten to be 3 hours because of saving time for template extraction.Multiplex direct PCR might serve the detection of food-borne pathogens in food safety risk monitoring much better.

Clinical study of using spectral CT at the minimal dose of contrast medium in cranial CTA / 中国生化药物杂志

Objective To investigate the minimum dose of contrast medium when using spectral CT in cranial CTA. Methods Three groups of patients were required to undergo head CTA examination because of their condition,but different doses of contrast agents were injected.Among them,group 30 mL, group B, group 40mL, group C and group 50 mL were A. Results In group A,the lowest rate of excellent image (23.33%), B group, C group had higher rate of excellent image (93.33%, 96.67%), comparison of the data P<0.05; C group the highest rate of adverse reaction of contrast agent (P<0.05),data comparison P<0.05, A group, B group significantly adverse reactions of contrast agent there is no difference between the situation (P>0.05). Conclusion The use of 40 mL iodide contrast medium in the procedure of cranial CTA examination is effective and safe.

Clinical study of using spectral CT at the minimal dose of contrast medium in cranial CTA / 中国生化药物杂志

Objective To investigate the minimum dose of contrast medium when using spectral CT in cranial CTA. Methods Three groups of patients were required to undergo head CTA examination because of their condition,but different doses of contrast agents were injected.Among them,group 30 mL, group B, group 40mL, group C and group 50 mL were A. Results In group A,the lowest rate of excellent image (23.33%), B group, C group had higher rate of excellent image (93.33%, 96.67%), comparison of the data P<0.05; C group the highest rate of adverse reaction of contrast agent (P<0.05),data comparison P<0.05, A group, B group significantly adverse reactions of contrast agent there is no difference between the situation (P>0.05). Conclusion The use of 40 mL iodide contrast medium in the procedure of cranial CTA examination is effective and safe.

Evaluating the current protocol of influenza A (H1N1) based on the epidemic situations of Zhengzhou,a middle-sized city in China / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>From May 2009-January 2010, a total of 3768 biosamples were tested for influenza A (H1N1) infection at Zhengzhou center for disease control and prevention, China. 1452 cases were laboratory confirmed H1N1 infection and 2316 were considered suspected victims. To evaluate the current protocol of influenza A (H1N1) based on the epidemic situations of Zhengzhou, relationships among features were explored and whether additional clinical characteristics should be part of H1N1 diagnosis protocols were determined.</p><p><b>METHODS</b>Both clinical and epidemiologic findings as well as statistical analyses were described in this article. Test for independence between features related to the disease diagnosis has been proposed. Furthermore, logistic regression was carried out to measure the association among features and latent class analysis was performed to identify additional crucial features in laboratory confirmed H1N1 by building various latent models with different combinatorial features.</p><p><b>RESULTS</b>The mean generation time for H1N1 was estimated as 3.59 +/- 1.41 days (range = 2.01-7.26). The estimated infection rate was 0.258 +/- 0.088 3, and reproduction number was 1.94 (95% CI = 1.12-3.18). Our results revealed that the six features, including molecular detections using three separate primer/probe sets, gender, age and temperature, are all associated with clinical diagnosis of H1N1, and that three separate primer/probe sets for laboratory confirmed H1N1, age and temperature are associated with each other.</p><p><b>CONCLUSION</b>Additional clinical features applied into the H1N1 diagnosis with current three primers/probe sets can increase the diagnostic efficiency.</p>

Effects of dexamethasone on expression of nuclear factor kappa B/inhibitor kappa B alpha and apoptosis of lens epithelial cells / 中华实验眼科杂志

Background Researches demonstrated that the long-term application of glucocorticoids can induce cataract. However, its molecular mechanism is unclear. Objective Present study was to investigate the effects of dexamethasone on the regulation of nuclear factor kappa B( NF-κB)/ inhibitor kappa B alpha( IκBα) line on human lens epithelial cells (LECs) and the LECs apoptosis. Methods Human LECs line(HLE2B3) were cultured and passaged in DMEM containing 20% fetal bovine serum and treated by different concentrations of dexamethasone(0. 01,0. 1,1,10,100 μmol/L) for 24,36 and 48 hours respectively. The LECs cultured in free-serum DMEM without dexamethasone were as blank control group. The expressions of IκBo: in the LECs were examined by reverse transcription polymerase chain reaction ( RT-PCR) and Western blot, and the expressions of NF-κB neucleoprotein in LECs were detected by Western blot after exposure to dexamethasone. The apoptosis rate of LECs was determined by flow cytometer. Results Agarose gel electrophoresis showed that the amplified gene fragment was coincident to designed one. The expressing level of NF-κB neucleoprotein in LECs was significantly lowed with the increase of dexamethasone concentration ( F = 36. 077 , P = 0. 004 ) , and that of IkBo: was evidently ascended ( F = 35. 741 ,P = 0. 002). In the same concentration of dexamethasone group,the expression of NF-κB in LECs showed the considerable alteration in different duration after treated of dexamethasone with the lowest expressing level in 36 hours, and significant differences were found in the expressing level between 24 hours and 36 hours ( P = 0. 002) and between 24 hours and 48 hours (P = 0. 01). The differences of expression of IκBá in LECs appeared the same pattern to NF-κB neucleoprotein. Flow cytometry showed that the apoptosis rate of LECs was obviously enhanced after action of dexamethasone in a dose-dependent manner, showing a significant difference among different groups ( F = 73. 261, P = 0.001). Conclusion It is implied that dexamethasone results in the pathogenesis and development of glucocorticoid cataract by up-regulating the expression of IκBα in LECs and suppressing the activity of NF-κB and herein induce the apoptosis of LECs at concentration-and time-dependent manner. This might be one of cellular and biological mechanisms of glucocorticoid cataract formation.

Sequence analysis of the phosphoprotein gene of peste des petits ruminants virus of Chinese origin / 病毒学报

The nucleotide sequences of P gene from a field strain of peste des petits ruminants virus (PPRV) ("China/Tib/Gej/07-30") was firstly determined. The P gene is 1,655 nucleotides long with two overlapping open reading frames (ORFs). The first ORF is 1530 nucleotides long and would produce P protein of 509 amino acid residues. The second ORF is 534 nucleotides long and would produce C protein of 177 amino acid residues. The first ORF produces a second mRNA transcript of 897 nucleotides long with an extra G nucleotide at position 751. Translation from this mRNA would produce V protein of 298 amino acid residues. The nucleotide and deduced amino acid sequence were compared with the homologous region of other PPRV isolates. At the amino acid level, the "China/Tib/Gej/07-30" shares homology of 86.10%-97.3%, 84.3%-94.9%, and 82.9%-96.3% for P, C, and V proteins respectively. Several sequence motifs in the P genes were identified on the basis of conservation in the PPRVs and the morbilliviruses.

Sequence analysis of the matrix protein and fusion protein genes of a field peste des petits ruminants virus strain from Tibet, China / 病毒学报

The nucleotide sequences of M and F genes from a field strain of peste des petits ruminants virus (PPRV) ("China/Tib/Gej/07-30") was firstly determined. The M gene was 1 483 nucleotides in length with a single open reading frame (ORF), encoding a protein of 335 amino acids. The F gene was 2411 nucleotides in length, encoding a protein of 546 amino acids. The resulting nucleotide sequence and the deduced amino acid sequences were compared with the homologous regions of other PPRV isolates. The nucleotide sequences of M and F genes of the "China/Tib/Gej/07-30" was 92.4%-97.7% and 85.5%-96.1% identical to other PPRV isolates, respectively, while a homology of 97.0%-98.2% and 94.3%-98.2% could be observed at the amino acids level respectively. Several sequence motifs in the M and F genes had been identified on the basis of conservation in the PPRVs and the morbilliviruses. The 3' untranslated region of M gene was 443 nucleotides in length with 82.4%-93.5% identical to other PPRV isolates. The 5' untranslated region of F gene was 634 nucleotides in length with 76.2%-91.7% identical to other PPRV isolates.

Sequence analysis of norovirus CHN02/LZ35666 from Lanzhou city of China based on RdRp and VP1 genes / 病毒学报

Sequence analysis of a new norovirus (NV) isolated from Lanzou city of China was performed based on partial sequence of RNA dependent RNA polymerase (RdRp) and complete capsid protein (VP1) gene. The isolated strain CHN02/LZ35666 shared high sequence homology with GII-4 NVs. Nucleotide homologies of RdRp region and encoded capsid protein region were 90.4% -- 98.6% and 89.8% -- 95.7% , respectively, while amino acid homology of capsid protein region was 94.4% -- 97.4%. The analysis of GDD motif in RdRp region indicated this GDD motif of Lanzhou strain differed from those of the GII-4 predominant epidemic strains. Lanzhou strain formed an independent branch in GII-4 cluster in the phylogenetic tree based on nucleotide sequence of RdRp region and amino acid sequence of capsid protein. Sequence alignment revealed a mutation at the fourth key site of the receptor-binding interface in the strains isolated after 2002 compared with those of previous strains suggesting a possible change of binding pattern to HBGAs receptors.

Comparison of bicyclol therapy for patients with genotype B and C of hepatitis B virus / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To compare the efficacy of bicyclol tablets on patients infected with hepatitis B virus between genotype B and C.</p><p><b>METHODS</b>70 patients with chronic viral hepatitis B were selected. The patients divide into two groups: HBV genotypes B (26 cases) and HBV genotypes C (other 44 cases). All patients received bicyclol tablets orally 150 mg daily (50mg, tid, po) for 24 weeks. The efficacy were observed after 12 weeks and 24 weeks.</p><p><b>RESULTS</b>After treatment for 24 weeks, the serum aminotransferase were decreased obviously, and HBV DNA levels turn to be negative with 19.2 percent (genotype B group) and 15.9 percent (genotype C group), respectively. The difference was not statistically significant between HBV genotype B and C.</p><p><b>CONCLUSION</b>Bicyclol not only has hepatoprotective activity but also inhibited virus replication in patients infected with HBV. The difference of the response to bicyclol therapy between HBV genotypes B and C was not statistically significant.</p>

Isolation and identification of arboviruses in Hebei Province / 中华实验和临床病毒学杂志

<p><b>BACKGROUND</b>To study the arboviruses carried by mosquitoes collected in Hebei Province.</p><p><b>METHODS</b>Samples were collected from mosquito active sites and stored in liquid nitrogen till use. Pools of 20 to 30 mosquitoes were ground after sterilization, centrifugal supernant was inoculated onto C6/36 cell, cytopathic effect was observed for three sequential passages. Positive isolates were identified by IFA and RT-PCR.</p><p><b>RESULTS</b>Totally 1310 mosquitoes were collected from two villages of She county, Hebei province. They were divided into 46 pools and ground respectively. Thirteen positive isolates were obtained. Two isolates reacted with alphaviral antibodies and were amplified by alphaviral primers, nucleotide sequence showed the highest homology (98%) to Getah virus (AY702913.1), so the two isolates were identified as Getah virus.</p><p><b>CONCLUSION</b>Getah virus was isolated from mosquitoes in Hebei Province. This is the first report of isolating Getah virus from inland of China.</p>

Sonographical features of the epididymis tuberculosis with caseous necrosis / 中华男科学杂志

<p><b>OBJECTIVE</b>To analyse sonographical features of epididymis tuberculosis with caseous necrosis and improve the accuracy of ultrasonic diagnosis.</p><p><b>METHODS</b>Ultrasonic features of 12 cases with epididymis tuberculosis with caseous necrosis, confirmed by surgical pathology, was retrospectively analysed: size, location, echogenicity, degree of blood flow in the lesion.</p><p><b>RESULTS</b>In all 12 patients, the number of purified protein derivative test (PPD) presenting the positive reached to 67%, while cases with the pulmonary tuberculosis was 42%. The epididymal tuberculosis with caseous necrosis could present images of the whole or focal epididymal enlargement and abnormal shape. The lesions internal echoes were irregular, inhomogeneous increased isoechoic echogenicity or hypoechoic, including bad-defined, irregular, homogeneous hypoechoic or anechoic. In lesional location, 4 out of 12 patients (33%) possessed richly color blood flow in Color Doppler Power Imaging(CDPI). The testicular enlargement with increased internal echoes and the testicular hydroceles were partly (58%, 7/12) observed by ultrasound. There was no varicocele, scrotal wall thickening and inguinal lymph node enlargement.</p><p><b>CONCLUSION</b>The High Frequency Color Doppler images is helpful for the diagnosis and differential diagnosis of epididymis tuberculosis with caseous necrosis.</p>