Video-assisted thoracoscopic extended thymectomy and transsternal extended thymectomy for treatment of myasthenia gravis: a case-control study / 南方医科大学学报

<p><b>OBJECTIVE</b>To compare the therapeutic effects of video-assisted thoracoscopic extended thymectomy (VATET) and transsternal extended thymectomy (TET) for myasthenia gravis (MG).</p><p><b>METHOD</b>This study included 21 patients undergoing VATET through the "three holes" approach on the right chest and 32 undergoing TET with sternum dissection. The thymus was excised and the anterior mediastinum adipose tissue removed in both groups.</p><p><b>RESULTS</b>VATET was associated with reduced intraoperative blood loss and longer operative time without the use of postoperative analgesics; very few patients were admitted into the intensive care unit (ICU), showing significant differences from the TET group (P<0.05). No significant difference was found between the two groups in tracheal tube removal time, length of stay in ICU, closed thoracic drainage removal time, and postoperative hospital stay, total hospital stay, postoperative complications, total hospitalization costs, or the rate of remission and improvement (P>0.05).</p><p><b>CONCLUSIONS</b>Compared with TET, VATET requires only a small incision without leaving metal foreign body in the body, and the patients experience less postoperative pain and rapid recovery, with similar mid- and long-term clinical outcomes.</p>

Detection of Banna virus-specific nucleic acid with TaqMan RT-PCR assay / 中华实验和临床病毒学杂志

<p><b>BACKGROUND</b>To develop a rapid and more sensitive TaqMan RT-PCR assay specific for Banna virus.</p><p><b>METHODS</b>Total RNA of strains of Banna virus and other arboviruses were extracted and reverse transcribed to cDNAs. With the cDNAs as templates, the TaqMan RT-PCR assay was developed and optimized on ABI 7300 apparatus for specific-detection of Banna virus. The sensitivity, specificity and the possibility of this assay to detect Banna virus in pools of mosquitoes. Meanwhile, human sera samples added with different dilutions of Banna virus culture medium supernatant were evaluated.</p><p><b>RESULTS</b>All the collected 12 Banna virus strains in our country were detected as positive, while the results of other arboviruses such as Japanese encephalitis virus (JEV), Batai virus, Sindbis virus and Orbiviruses were negative. This assay was more than 100 times sensitive than that of traditional PCR. About 0.5 CCID-50 of Banna virus in 50 microl samples could be detected with this assay. The sensitivity decreased by about 10 times when the dsRNA was denatured in 65 degrees C instead of 99 degrees C, and also when the virus seeds were kept at room temperature for 1 day or 1 week at 4 degrees C. There was no significant difference in the results of Banna virus detection between artificial human serum samples and positive control virus. Six were detected as positive for Banna virus-specific nucleic acids in 112 pools of mosquitoes, while 3 was positive with virus isolation.</p><p><b>CONCLUSION</b>The Banna virus-specific TaqMan RT-PCR assay was proved to be highly sensitive, specific and showed a good reproducibility; the assay may be applied for surveillance of this virus in clinical samples and pools of mosquitoes screening.</p>

Isolation and identification of Japanese encephalitis virus in Liaoning Province / 中华实验和临床病毒学杂志

<p><b>BACKGROUND</b>To study arboviruses carried by mosquitoes in Liaoning Province in 2002.</p><p><b>METHODS</b>Totally 4927 mosquitoes were collected from Liaoning Province in July 2002. Virus strains were isolated by inoculating BHK-21, C6/36 and Vero cells. The newly isolated strains were identified by serological (ELISA and IFA) and molecular methods (Real-Time PCR and RT-PCR).</p><p><b>RESULTS</b>Two strains were isolated from mosquitoes causing cytopathogenic effect (CPE) on cells and were fatal to suckling mice. Serological tests showed that both were positive for the antibody to JEV. The PrM and E gene were cloned and sequenced. The phylogenetic analysis showed that the new isolates belonged to genotype I, JEV. Sequence analysis showed that the homology of nucleotide sequences and amino acid (AA) sequences between the two strains was 100%. Compared with the nucleotide sequences between the two strains and the standard JEV vaccine strain SA14-14-2, the difference was up to 4.11%, and the difference of AA was 0.6%.</p><p><b>CONCLUSION</b>Two strains of JEV were isolated and identified in Liaoning province, both belonged to genotype I JEV.</p>

Establishment of TaqMan PCR detection method for rabies virus / 中华流行病学杂志

<p><b>OBJECTIVE</b>To establish a molecular diagnostic method for rabies virus(RV) based on TaqMan PCR.</p><p><b>METHODS</b>BaseD on the rabies virus nucleoprotein gene sequences published in GenBank, RV specific primers and probe were designed by Primer Premier 5.0. The primers and probe were optimized and the sensitivity, specificity,and reproducibility of the system were tested. Quantitative standard curve of RV TaqMan PCR was established. Some RV samples were detected using this system.</p><p><b>RESULTS</b>The optimized primers and probe were 0.6 micromol/L and 0.2 micromol/L. Reproducibility test showed that coefficient variables were all less than 5% in 4 different system. Quantification standard curve based on the genomic copy was drawn. RV detection using the established method proved that TaqMan PCR was more sensitive and easier performed than traditional RT-PCR.</p><p><b>CONCLUSION</b>TaqMan PCR for RV detection had been established, which was more sensitive and specific than the general RT-PCR.</p>

Isolation and identification of arboviruses from mosquito pools in Yunnan Province / 中华实验和临床病毒学杂志

<p><b>BACKGROUND</b>To survey arboviruses in Yunnan province.</p><p><b>METHODS</b>Mosquitoes were collected from Yunnan Province in 2002 and 2004. Virus strains were isolated by the inoculation of homogenates of the mosquitoes onto BHK cell line. The isolated strains and their molecular biological characteristics were identified by real-time PCR, reverse transcription-polymerase chain reaction (RT-PCR) and immunofluorescent antibody technique.</p><p><b>RESULTS</b>Twelve strains of viruses producing CPE in BHK cells were isolated from 4810 mosquitoes. All the 12 isolates were identified to be Japanese encephalitis viruses. Genotype analysis showed the new virus (DL-0437 strain) belonged to genotype III.</p><p><b>CONCLUSION</b>Twelve strains of Japanese encephalitis viruses were isolated from mosquito pools collected in Yunnan. It was the first isolation of genotype III Japanese encephalitis viruses in Yunnan Province in recent years.</p>

Molecular biological identification of Batai virus isolated in China / 中华实验和临床病毒学杂志

<p><b>BACKGROUND</b>To study the molecular characteristics of YN92-4 strain isolated from mosquitoes in Yunnan Province and define its classification.</p><p><b>METHODS</b>The S segment of YN92-4 strain was amplified and sequenced by 2 different sets of primers. The phylogenic tree of S fragment was constructed by Phylip bio-software. The amino acid sequences of N and NSs proteins were also studied.</p><p><b>RESULTS</b>YN92-4 strain could be amplified by 2 sets of primers respectively, S segment showed a highest homology with Batai virus (X73464), reached 96.4%, the homology of protein N and NSs amio-acid sequence with Batai virus was 99.1% and 98% respectively.</p><p><b>CONCLUSION</b>The YN92-4 strain belongs to Batai virus, this is the first report of molecular biological identification of Batai virus in China.</p>

Detection of Sindbis virus-specific nucleic acid with SYBR GREEN I real time PCR assay / 中华实验和临床病毒学杂志

<p><b>BACKGROUND</b>To develop a rapid, specific and sensitive method for detecting Sindbis virus (SINV) with SYBR GREEN I real time PCR.</p><p><b>METHODS</b>Total RNA of strains of Sindbis virus and a related virus were extracted and reverse transcribed to cDNAs. With the cDNAs as template, the SYBR GREEN I real time PCR assay was developed and optimized on ABI 7300 apparatus for specific-detection of Sindbis virus, and the sensitivity, specificity and reproducibility were evaluated.</p><p><b>RESULTS</b>For the PCR, 55 degrees C was chosen as the optimal anneal temperature and 0.5 micromol/L as the optimal primer concentration. Using this method, all the selected SINV were detected as positive, while the results of control arboviruses such as Geta virus, Japanese encephalitis virus (JEV), Batai virus, Seadornavirus Orbiviruses and synthesized WEEV cDNA were negative. With this system, 0.1 PFU/ml SINV cDNA could be detected; The sensitivity of this assay was about 100 times higher than standard RT-PCR. All the results were reproducible within two compatible tests, and the stability of the detection system was very good. The test results of simulated infection human serum samples showed that human serum had no obvious interference with this system. With this system, 6 of 151 clinical samples with unknown fever or encephalitis were determined as positive.</p><p><b>CONCLUSION</b>The developed SYBR GREEN I real time PCR assay for detecting Sindbis virus was highly sensitive, specific and showed a good reproducibility and stability. It is our belief that the present method can be further used in clinic sample to verify its stringency.</p>