Complementing Tissue Testing With Plasma Mutation Profiling Improves Therapeutic Decision-Making for Patients With Lung Cancer.

BACKGROUND: Tissue biopsy is an integral part of the diagnostic approach to lung cancer. It is however invasive and limited by heterogeneity. Liquid biopsies may complement tissue testing by providing additional molecular information and may be particularly helpful in patients from whom obtaining sufficient tissue for genomic profiling is challenging. METHODS: Patients with suspected lung cancer (n = 71) were prospectively recruited. Blood and diagnostic tissue samples were collected within 48 h of each other. Plasma cell-free DNA (cfDNA) testing was done using an ultrasensitive amplicon-based next-generation sequencing (NGS) panel (plasma NGS testing). For cases diagnosed as non-small cell lung carcinoma (NSCLC) via histology or cytology, targeted testing for epidermal growth factor receptor (EGFR) mutations was performed using tissue biopsy samples (tissue EGFR testing), where available. Concordance of clinically actionable mutations between methods and sample types was assessed. RESULTS: For confirmed NSCLC cases (n = 54), tissue EGFR test results were available only for 70.3% (38/54) due to sample inadequacies, compared to blood samples for 98.1% (53/54) cases. Tissue EGFR testing identified sensitizing EGFR (L858R or exon 19 deletion) mutation in 31.6% (12/38) of cases. Plasma NGS identified clinically actionable mutations in 37.7% (20/53) of cases, including EGFR mutations in two cases with no tissue EGFR results, and mutations in KRAS, BRAF, and MET. The overall sensitivity of sensitizing EGFR mutation detection by plasma NGS was 75% (9/12), and specificity was 100% (25/25) in patients tested in both tissue EGFR and plasma NGS (n = 37). In this cohort of patients, tissue EGFR testing alone informed clinical decisions in 22.2% (12/54) of cases. Adding plasma NGS to tissue EGFR testing increased the detection rate of actionable mutations to 42.6% (23/54), representing a 1.9-fold increase in clinically relevant findings. The average turnaround time of plasma NGS was shorter than standard tissue testing (10 vs. 29.9 days, p < 0.05). CONCLUSIONS: In the first-line setting, plasma NGS was highly concordant with tissue EGFR testing. Plasma NGS increases the detection of actionable findings with a shorter time to results. This study outlines the clinical utility of complementary plasma mutation profiling in the routine management of lung cancer patients.

An efficient TILLING platform for cultivated tobacco.

Targeting-induced local lesions in genomes (TILLING) is a powerful reverse-genetics tool that enables high-throughput screening of genomic variations in plants. Although TILLING has been developed for many diploid plants, the technology has been used in very few polyploid species due to their genomic complexity. Here, we established an efficient capillary electrophoresis-based TILLING platform for allotetraploid cultivated tobacco (Nicotiana tabacum L.) using an ethyl methanesulfonate (EMS)-mutagenized population of 1,536 individuals. We optimized the procedures for endonuclease preparation, leaf tissue sampling, DNA extraction, normalization, pooling, PCR amplification, heteroduplex formation, and capillary electrophoresis. In a test screen using seven target genes with eight PCR fragments, we obtained 118 mutants. The mutation density was estimated to be approximately one mutation per 106 kb on average. Phenotypic analyses showed that mutations in two heavy metal transporter genes, HMA2S and HMA4T, led to reduced accumulation of cadmium and zinc, which was confirmed independently using CRISPR/Cas9 to generate knockout mutants. Our results demonstrate that this powerful TILLING platform (available at http://www.croptilling.org) can be used in tobacco to facilitate functional genomics applications.

Alterations in the transmural gradient of ventricular repolarization with different pacing sites in normal and heart failure canines / 生理学报

Alterations of the transmural gradient of repolarization may contribute to the increase of transmural dispersion of repolarization and ventricular arrhythmias. The transmural gradient of repolarization may play an important role in sudden death associated with left ventricular epicardial pacing. To investigate the changes of transmural gradient dispersion of ventricular repolarization with different pacing sites in heart failure (HF) canines, 8 mongrel dogs were randomized into healthy group and HF group (n = 4). We mapped the monophasic action potential duration (MAPD) in the subendocardial, subepicardial and mid-myocardial layers of the left ventricle (LV) in canines of healthy and HF groups during right atrium (RA) pacing, right ventricular apical endocardial (RV) pacing, left ventricular lateral epicardial (LV) pacing and biventricular (Biv) pacing respectively. The results showed that in the healthy group, the MAPDs were significantly different among the three layers during RA pacing (all P 0.05). In the HF group, the MAPDs in all three layers were prolonged compared with those in the same locations in the healthy group (all P 0.05). By MAP recording with our new mapping electrode, we found a transmural MAPD gradient among the three layers of the LV during RA pacing and the gradient between the subendocardial and subepicardial layers vanished during RV, LV or Biv pacing in healthy dogs. In contrast, there was no transmural MAPD gradient during RA, RV, LV or Biv pacing in HF dogs. These results are helpful to understand the mechanism of ventricular arrhythmias in patients with HF.

Routine semen parameters and sperm morphological index of infertile males with partial globozoospermia / 中华男科学杂志

Objective@#To investigate the routine semen parameters and sperm morphological indexes of the patients with partial globozoospermia (PGZ).@*METHODS@#We included in this study 100 infertile males with PGZ and another 180 non-PGZ infertile men as controls. According to the proportion of round-headed sperm (RHS) in the semen, we classified the PGZ males into five subgroups: 25%－40%, 41%－55%, 56%－70%, 71%－85%, and 86%－99% RHS. We obtained sperm concentration, total sperm motility, the percentage of progressively motile sperm, teratozoospermia index (TZI), and sperm deformity index (SDI) from the subjects and compared them among different groups.@*RESULTS@#Statistically significant differences were found between the PGZ patients and non-PGZ controls in total sperm motility (［35.76±24.88］% vs ［62.03±10.20］%, P0.05), sperm concentration (［46.01±40.38］ vs ［54.00±25.15］ ×106/ml, P>0.05), or TZI (1.35±0.11 vs 1.34±0.54, P>0.05). There were also significant differences among the five PGZ subgroups in total sperm motility, progressive sperm motility, normal sperm morphology, TZI, and SDI (P0.05). Morphologically, the sperm head changed from heterogeneous to homogeneous with the increased proportion of round-headed sperm.@*CONCLUSIONS@#Different proportions of round-headed sperm are closely related to routine semen parameters and sperm morphological index in PGZ patients, which can help clinicians choose the proper assisted reproductive technology and predict the rate of fertilization for infertile males.

Risk factors of leptospirosis in Liping County,Guizhou Province, China——an expanded survey in general population and rodents / 中国人兽共患病学报

Leptospirosis has long been a sporadic zoonosis in Liping County.This study aims to explore the risk factors of leptospirosis infection in general population,and the role rodents play in transmission by an expanded investigation of two deceased cases.Serology survey and demographic investigation were carried out among the general population of two epidemic areas where two deseased cases had been reported,and one extended area where no leptospirosis cases had been diagnosed.Local rodents' kidney samples were collected for Leptospira isolation.In total 44 people (23.4％) tested positive by microscopic agglutination test (MAT) for four strains of Leptospira.Seropositive rate of the extended area was significantly higher (53.3 ％,24/45) than epidemic area (14.0％,20/143).Positive rate increased with age.Other factors such as occupation,gender showed no significance.The proportion of Apodemus agrarius in the captured rodents and the total germ-carrying rates showed no significant difference in two areas.Long-term rodent carrier surveillance is critical to provide valuable information and locate high risk areas,facilitating prophylactic measures.Vaccination was recommended in high risk areas if economical feasibility and demand were proved.

A model to predict the risk of lethal nasopharyngeal necrosis after re-irradiation with intensity-modulated radiotherapy in nasopharyngeal carcinoma patients.

BACKGROUND: For patients with nasopharyngeal carcinoma (NPC) who undergo re-irradiation with intensity-modulated radiotherapy (IMRT), lethal nasopharyngeal necrosis (LNN) is a severe late adverse event. The purpose of this study was to identify risk factors for LNN and develop a model to predict LNN after radical re-irradiation with IMRT in patients with recurrent NPC. METHODS: Patients who underwent radical re-irradiation with IMRT for locally recurrent NPC between March 2001 and December 2011 and who had no evidence of distant metastasis were included in this study. Clinical characteristics, including recurrent carcinoma conditions and dosimetric features, were evaluated as candidate risk factors for LNN. Logistic regression analysis was used to identify independent risk factors and construct the predictive scoring model. RESULTS: Among 228 patients enrolled in this study, 204 were at risk of developing LNN based on risk analysis. Of the 204 patients treated, 31 (15.2%) developed LNN. Logistic regression analysis showed that female sex (P = 0.008), necrosis before re-irradiation (P = 0.008), accumulated total prescription dose to the gross tumor volume (GTV) ≥145.5 Gy (P = 0.043), and recurrent tumor volume ≥25.38 cm(3) (P = 0.009) were independent risk factors for LNN. A model to predict LNN was then constructed that included these four independent risk factors. CONCLUSIONS: A model that includes sex, necrosis before re-irradiation, accumulated total prescription dose to GTV, and recurrent tumor volume can effectively predict the risk of developing LNN in NPC patients who undergo radical re-irradiation with IMRT.

Neoadjuvant chemotherapy plus intensity-modulated radiotherapy versus concurrent chemoradiotherapy plus adjuvant chemotherapy for the treatment of locoregionally advanced nasopharyngeal carcinoma: a retrospective controlled study.

BACKGROUND: In the era of intensity-modulated radiotherapy (IMRT), the role of neoadjuvant chemotherapy (NAC) for locoregionally advanced nasopharyngeal carcinoma (NPC) is under-evaluated. The aim of this study was to compare the efficacy of NAC plus IMRT and concurrent chemoradiotherapy (CCRT) plus adjuvant chemotherapy (AC) on locoregionally advanced NPC. METHODS: Between January 2004 and December 2008, 240 cases of locoregionally advanced NPC confirmed by pathologic assessment in Sun Yat-sen University Cancer Center were reviewed. Of the 240 patients, 117 received NAC followed by IMRT, and 123 were treated with CCRT plus AC. The NAC + IMRT group received a regimen that included cisplatin and 5-fluorouracil (5-FU). The CCRT + AC group received cisplatin concurrently with radiotherapy, and subsequently received adjuvant cisplatin and 5-FU. The survival rates were assessed by Kaplan-Meier analysis, and the survival curves were compared using a log-rank test. Multivariate analysis was conducted using the Cox proportional hazard regression model. RESULTS: The 5-year overall survival (OS), locoregional relapse-free survival (LRRFS), distant metastasis-free survival (DMFS), and disease-free survival (DFS) were 78.0, 87.9, 79.0, and 69.8%, respectively, for the NAC + IMRT group and 78.7, 84.8, 76.2, and 65.6%, respectively, for the CCRT + AC group. There were no significant differences in survival between the two groups. In multivariate analysis, age (<50 years vs. ≥50 years) and overall stage (III vs. IV) were found to be independent predictors for OS and DFS; furthermore, the overall stage was a significant prognostic factor for DMFS. Compared with the CCRT + AC protocol, the NAC + IMRT protocol significantly reduced the occurrence rates of grade 3-4 nausea-vomiting (6.5 vs. 1.5%, P = 0.023) and leukopenia (9.7 vs. 0.8%, P = 0.006). CONCLUSIONS: The treatment outcomes of the NAC + IMRT and CCRT + AC groups were similar. Distant metastasis remained the predominant mode of treatment failure.

Central venous catheter-related thrombosis in senile male patients: New risk factors and predictors / 华中科技大学学报（医学）（英德文版）

Central venous catheterization (CVC)-related venous thrombosis is a common but serious clinical complication, thus prevention and treatment on this problem should be extensively investigated. In this research, we aimed to investigate the incidence rate of CVC-related venous thrombosis in senile patients and give a further discussion on the related risk factors and predictors. A total of 324 hospitalized senile male patients subjected to CVC were selected. Retrospective investigation and analysis were conducted on age, underlying diseases, clinical medications, catheterization position and side, catheter retention time, and incidence of CVC-related venous thrombosis complications. Basic laboratory test results during catheterization and thrombogenesis were also collected and analyzed. Among the 324 patients, 20 cases (6.17%) of CVC-related venous thrombosis were diagnoseds. The incidence rate of CVC-related venous thrombosis in subclavian vein catheterization was significantly lower than that in femoral vein catheterization (P<0.01) and that in internal jugular vein catheterization (P<0.05). No statistically significant difference was found between femoral vein catheterization and internal jugular vein catheterization (P<0.05). Previous venous thrombosis history (P<0.01), high lactate dehydrogenase level (P<0.01), low high-density lipoprotein (HDL) level (P<0.05), and low albumin level (P<0.05) were found as risk factors or predictors of CVC-related venous thrombosis in senile male patients. Subclavian vein catheterization was the most appropriate choice among senile patients to decrease the incidence of CVC-related venous thrombosis. Previous venous thrombosis history, high lactate dehydrogenase level, low HDL level, and low albumin level were important risk factors in predicting CVC-related venous thrombosis.

Central venous catheter-related thrombosis in senile male patients: New risk factors and predictors / 华中科技大学学报（医学）（英德文版）

Central venous catheterization (CVC)-related venous thrombosis is a common but serious clinical complication, thus prevention and treatment on this problem should be extensively investigated. In this research, we aimed to investigate the incidence rate of CVC-related venous thrombosis in senile patients and give a further discussion on the related risk factors and predictors. A total of 324 hospitalized senile male patients subjected to CVC were selected. Retrospective investigation and analysis were conducted on age, underlying diseases, clinical medications, catheterization position and side, catheter retention time, and incidence of CVC-related venous thrombosis complications. Basic laboratory test results during catheterization and thrombogenesis were also collected and analyzed. Among the 324 patients, 20 cases (6.17%) of CVC-related venous thrombosis were diagnoseds. The incidence rate of CVC-related venous thrombosis in subclavian vein catheterization was significantly lower than that in femoral vein catheterization (P<0.01) and that in internal jugular vein catheterization (P<0.05). No statistically significant difference was found between femoral vein catheterization and internal jugular vein catheterization (P<0.05). Previous venous thrombosis history (P<0.01), high lactate dehydrogenase level (P<0.01), low high-density lipoprotein (HDL) level (P<0.05), and low albumin level (P<0.05) were found as risk factors or predictors of CVC-related venous thrombosis in senile male patients. Subclavian vein catheterization was the most appropriate choice among senile patients to decrease the incidence of CVC-related venous thrombosis. Previous venous thrombosis history, high lactate dehydrogenase level, low HDL level, and low albumin level were important risk factors in predicting CVC-related venous thrombosis.

The impact of baseline serum C-reactive protein and C-reactive protein kinetics on the prognosis of metastatic nasopharyngeal carcinoma patients treated with palliative chemotherapy.

BACKGROUND: The aim of this study was to determine whether baseline C-reactive protein (CRP) levels and CRP kinetics predict the overall survival in metastatic nasopharyngeal carcinoma (mNPC) patients. METHODS: A total of 116 mNPC patients from January 2006 to July 2011 were retrospectively reviewed. Serum CRP level was measured at baseline and thereafter at the start of each palliative chemotherapy cycle for all patients. RESULTS: Patients with higher values of baseline CRP (≥ 3.4 mg/L) had significantly worse survival than those with lower baseline CRP values (< 3.4 mg/L). Patients were divided into four groups according to baseline CRP and CRP kinetics: (1) patients whose CRP < 3.4 mg/L and never elevated during treatment; (2) patients whose CRP < 3.4 mg/L and elevated at least one time during treatment; (3) patients whose CRP ≥ 3.4 mg/L and normalized at least one time during treatment; and (4) patients whose CRP ≥ 3.4 mg/L and never normalized during treatment. The patients were further assigned to non-elevated, elevated, normalized, and non-normalized CRP groups. Overall survival rates were significantly different among the four groups, with three-year survival rates of 68%, 41%, 33%, and 0.03% for non-elevated, elevated, normalized, and non-normalized CRP groups respectively. When compared with the non-elevated group, hazard ratios of death were 1.69, 2.57, and 10.34 in the normalized, elevated, and non-normalized groups (P < 0.001). CONCLUSIONS: Baseline CRP and CRP kinetics may be useful to predict the prognosis of metastatic NPC patients treated with palliative chemotherapy and facilitate individualized treatment. A prospective study to validate this prognostic model is still needed however.

A prognostic model predicts the risk of distant metastasis and death for patients with nasopharyngeal carcinoma based on pre-treatment serum C-reactive protein and N-classification.

PURPOSE: Chronic inflammation plays an important role in nasopharyngeal carcinoma (NPC) development and progression. Aim of this study is to determine whether inflammation-related parameters predict distant metastasis in NPC patients. MATERIALS AND METHODS: 335 newly diagnosed non-metastatic NPC patients were recruited. The values of the C-reactive protein (CRP), lactate dehydrogenase, albumin, globulin, white blood cell and neutrophil at baseline were measured. RESULTS: Among the above six parameters, only CRP was independently associated with distant metastasis-free survival (DMFS). CRP concentration of advanced T-/TNM-classification patients was higher than those with early classification (P = 0.001). Higher-CRP (CRP ⩾ 2.46 mg/L) predicted shorter overall survival, disease-free survival and DMFS than lower-CRP (CRP < 2.46 mg/L). In a multivariable model, higher-CRP and advanced N-classification were independent predictors of distant metastasis. On the basis of these two parameters, a prognostic NC-model was developed as following: (1) low-risk (early N-classification and lower-CRP); (2) intermediate-risk (advanced N-classification or higher-CRP) and (3) the high-risk distant metastasis (advanced N-classification and higher-CRP). When compared with the low-risk group, the hazard ratios (HRs) for distant metastasis and death for the intermediate-/high-risk patients were 3.6/16.1 and 2.26/7.61, respectively (both P < 0.001). CONCLUSION: We developed a new prognostic model based on CRP and N-classification for predicting distant metastasis and death of NPC patients, which may facilitate patient counselling and individualised treatment.

Identification and genetic characteristics on the bacteria isolate from a case of human Brucellosis in Guizhou province / 中华流行病学杂志

<p><b>OBJECTIVE</b>A suspected Brucella (B.) strain(GZZA), isolated from a case of anti-Brucella antibody positive patient was identified and its' genetic characteristics was analyzed, to provide etiologic basis for the confirmation of patient in Guizhou province.</p><p><b>METHODS</b>Conventional methods and polymerase chain reaction(PCR)were used to identify the bacteria strain, with genetic characteristics analyzed by MLVA-16.</p><p><b>RESULTS</b>The bacteria strain was identified as B. melitensis biovar 3 under the conventional and PCR methods. Results from the MLVA-16 analysis indicated that the bacteria strain was closely clustered with B. melitensis biovar 3, and differences of repeated numbers at VNTR loci bruce42, bruce04, bruce09 and bruce16 were also displayed.</p><p><b>CONCLUSION</b>Both traditional and molecular methods to identify one bacteria strain isolated from the human patient as B. melitensis biovar 3 and the genetic characteristics of the strain was closely related to that of B. melitensis biovar 3. Differences of repeated numbers at part of VNTR loci were also showed. The results of this study provided etiologic evidences for the confirmation of Brucella infection of the patient, also providing scientific basis for the control and prevention of Brucellosis in Guizhou province.</p>

Detection of SALL4 mRNA expression in leukemia by real-time fluorescent quantitative PCR / 中国实验血液学杂志

This study was purposed to establish a real-time fluorescent quantitative PCR (FQ-PCR) for quantifying SALL4 mRNA and to investigate its expression in different types of leukemia patients. SALL4 mRNA expression were measured in 60 leukemia patients of different periods and 10 normal controls sequentially by FQ-PCR. The results showed that the expression of SALL4 mRNA in de novo leukemia patients and relapsed patients was higher than that in controls (P < 0.05), which was significantly decreased at complete remission (CR). In relapsed patients, the expression of SALL4 mRNA increased slightly higher than that in de novo leukemia group, but the difference was not statistically significant (P > 0.05). However, the expression of SALL4 mRNA was low in CLL, T-ALL and AML-M3. The expression pattern of BMI-1 was same as SALL4, and the expression of BMI-1 positively correlated with that of SALL4 in leukemia (r = 0.825, P < 0.01). It is concluded that the detection of SALL4 gene expression in acute and chronic leukemia by real-time gTR-PCR displays high sensitivity and specificity. SALL4 gene may be one of indicators for monitoring the therapeutic outcome of partial leukemia and minimal residual disease.

Molecular typing of Leptospira interrogans strains isolated from Rattus tanezumi in Guizhou Province, Southwest of China / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To identify and type three leptospires isolated from Rattus tanezumi in Guizhou Province by using three molecular techniques (PFGE, MLVA, and MLST), reveal the molecular characteristic of causative agents of local leptospirosis and evaluate these three molecular methods based on their detection resolution and efficiency.</p><p><b>METHODS</b>Three Leptospira strains were isolated from the kidney of Rattus tanezumi and cultured with EMJH medium. PFGE, MLVA, and MLST assays were applied to type the three strains isolated from Rattus tanezumi in Guizhou Province.</p><p><b>RESULTS</b>PFGE, MLVA, and MLST typing showed that the three leptospiral isolates matched with leptospiral serogroup Icterohaemorrhagiae serovar Lai. The findings of the genotyping methods were consistent. MLVA and MLST defined genotypes, whereas PFGE allowed the recognition of additional subgroups within the genotypes, and the findings of molecular typing were also consistent with those of traditional techniques.</p><p><b>CONCLUSION</b>Three leptospiral isolates from Guizhou Province matched with leptospiral serogroup Icterohaemorrhagiae serovar Lai, and PFGE, MLVA, and MLST, as reliable molecular techniques for identifying and typing of Leptospira interrogans, would contribute to the active surveillance, outbreak investigation and source tracking for leptospirosis in Guizhou Province.</p>

Analysis of the genetic differences in the nucleoprotein between rabies virus and its vaccine strains in Guizhou province from year 2005 to 2010 / 中华预防医学杂志

<p><b>OBJECTIVE</b>This study was to explore the differences in the nucleoprotein gene between rabies virus (RABV) and its vaccine strains in Guizhou province from year 2005 to 2010.</p><p><b>METHODS</b>Samples from 4 rabies patients and cerebral tissue samples of 28 rabies infected dogs were collected from different districts in Guizhou province between year 2005 and 2010. Direct Immunofluorescence Assay (DFA) and RT-nested PCR assay were applied to detect the overall length of N gene sequence. Meanwhile, based on the comparison between the homology and phylogenetic tree, the differences in N gene sequence between the prevalent RABV and the RABV vaccine strains collected from NCBI database in these years.</p><p><b>RESULTS</b>According to DFA and RT-nested PCR assay, the antigen and nucleic acid of the 21 dogs and 4 human samples were both confirmed positive; whose full length of N gene sequences were both 1353 bp. The homological analysis showed that the 25 strains of RABV virus and the RABV type I virus stored by GenBank database shared a high homology in N gene nucleotide and amino acid sequences, which were 89%-100% and 98%-100%, respectively. Besides, the homology between the 25 strains of RABV virus and its vaccines in nucleotide and amino acid sequences were separately 86%-95% and 96%-100%. The N gene of vaccines for livestock shared the highest homology with HEP-Flury strain in the nucleotide and amino acid, which were 88%-89% and 98%-99%, respectively. The vaccines for human use showed its greatest homology with the CTN strain in nucleotide (86%-100%) and amino acid (96%-100%). The phylogenetic tree analysis indicated that the 25 strains of RABV virus, RABV type I virus and the CTN vaccine strains constituted one individual cluster, which was least different from the CTN vaccine for human use.</p><p><b>CONCLUSION</b>The prevalent RABV virus, the vaccine HEP-Flury for livestock and the vaccine CTN for human use were found to be highly similar in N gene expression in Guizhou province from 2005 to 2010.</p>

Etiologic diagnosis and analysis of the first case of human brucellosis in Guizhou province / 中国地方病学杂志

Objective To etiologically diagnose and analyze a patient with suspected cases of brucellosis,and to provide a experimental basis for the confirmation of the first case of human brucellosis in Guizhou province.Methods Conventional and molecular techniques [genus specific Brucella surface protein 31 PCR (BCSP31-PCR)and Brucella suis species-specific PCR (AMOS-PCR)] were used to identify suspicious bacteria strains isolated from the suspected patient of brucellosis.Results The results showed that the Brucella suspicious colonies were identified as Brucella melitensis biotype 3 using conventional tests and were further identified as Brucella spp.by genus specific Brucella surface protein 31 PCR (BCSP31-PCR) and classified as Brucella melitensis with Brucella abortus,Brucella melitensis,Brucella ovis,Brucella suis species-specific PCR(AMOS-PCR).Conclusions Laboratory diagnostic results show that the bacteria strain isolated from the suspected patient of brucellosis is Brucella melitensis biotype 3.It is the first case of human brucellosis in Guizhou province.

Analysis on nucleoprotein gene sequence of 25 rabies virus isolates in Guizhou Province, China / 病毒学报

To analyze 25 nucleoprotein gene (N gene) sequences of rabies viruses circulating in Guizhou province during 2005-2010, China, and to explore the epidemic characteristics and the probable mutant of rabies in Guizhou Province. Rabies virus RNA in human brain tissues, human saliva, and domestic dog brain tissues derived from different prefectures of Guizhou Province were detected with RT-nested PCR, and the amplified products were then sequenced. Bioinformatics software was used to determine the genetic characteristics of these rabies viruses. The sequences of N gene of 25 Guizhou provincial isolates were identical with homogeny between 97.5% - 99.3% and 98.4% - 99.8% at nucleotide and deduced amino acid level, respectively, while the identities between them and isolated strains from other province of China were 88% - 99.1% and 88% - 99.7%. There were several amino acid substitutions in the nucleoprotein of 25 Guizhou isolates compared with the known genotype 1 isolates. The analysis of phylogenetic tree of 25 Guizhou isolates was demonstrated to be genetically divided into two groups, indicating that the virus presented a unique characteristics in geographic distribution and in a time dependent-manner. And phylogenetic tree of 25 Guizhou isolates and 7 genotype 1 strains isolated from other Province of China was also divided into two groups, which were further composed of several subgroups, respectively. From these observations, the rabies viruses derived from Guizhou province were still genotype 1. These isolates of rabies virus were diverged from the strains isolated from other provinces in both gene sequences and deduced amino acid sequences, and these divergences were characterized in geographic distribution and in a time-dependent manner.

Restitution of atrial repolarization and atrial fibrillation in canine atrium / 中华心血管病杂志

<p><b>OBJECTIVE</b>Electrical restitution was believed to be a determinant responsible for the stability of heart rhythm. Although numerous studies focused on the role of action potential duration restitution (APDR) in the initiation and maintenance of ventricular fibrillation (VF), the relationship between atrial APDR and atrial fibrillation (AF) has not been fully understood. This study aims to investigate the characteristics of APDR of left atrium (LA) and right atrium (Rs) in canines and the relevance to induction of AF.</p><p><b>METHODS</b>Monophasic action potential (MAP) was recorded from LA and RA in 14 canines using the MAP recording-pacing combination catheter. APDR, plotted as action potential duration (APD) on the preceding diastolic interval (DI), was assessed by use of programmed stimulation with a single extrastimulus (S1S2) at LA and RA. Episodes of AF were recorded and analyzed.</p><p><b>RESULTS</b>APD90 was significantly shorter in the LA than that in the RA [(157.4 +/- 43.5) ms vs. (170.9 +/- 37.9) ms, P < 0.05]. The mean slope of the APDR curve by S1S2 in the LA was significantly greater than that in the RA (1.3 +/- 0.4 vs. 0.9 +/- 0.3, P < 0.05). The incidence of induced AF was significantly higher in the LA than in the RA (11/18 vs. 7/18, P < 0.05).</p><p><b>CONCLUSIONS</b>The APDR and MAP characteristics are not uniform between atrium, which may be one of the important mechanisms responsible for the initiation of AF. Heterogeneity of APDR between LA and RA might create critical gradients or a dispersion of repolarization and substrate for re-entrant arrhythmias and vulnerability to AF.</p>

Percutaneous vertebroplasty and kyphoplasty for the treatment of thoracolumbar fractures in the elderly / 中国骨伤

<p><b>OBJECTIVE</b>To study the clinical effects of percutaneous vertebroplasty and kyphoplasty in the treatment of fresh thoracolumbar fractures in the elderly.</p><p><b>METHODS</b>From May 2004 to March 2009, among 29 patients (32 vertebras) with thoracolumbar fractures, 18 patients (21 vertebras) were treated with percutaneous vertebroplasty (PVP) and 11 patients (11 vertebras) were treated with percutaneous kyphoplasty (PKP). The visual analogue scale (VAS) and mobility were evaluated 3 days before and after operation. During follow-up period, patients were observed and evaluated in terms of relief of back pain and regaining of living ability. Complications such as bone cement leakage were analyzed.</p><p><b>RESULTS</b>All the patients got successful operation. The average operation time was 38.6 minutes for the 18 patients (21 vertebras) with PVP performed, and 3.2 ml (2-5 ml) bone cement was injected. For the 11 patients (11 vertebras) with PKP performed, the average operation time was 43 minutes, and an average of 3.8 ml (2-6 ml) bone cement was injected. The VAS decreased from preoperative (7.5 +/- 0.7) to (2.2 +/- 0.5) at the 3rd day after operation; the mobility scores of the patients decreased from preoperative (2.4 +/- 0.6) to (1.2 +/- 0.3) at the 3rd day after operation; the vertebral height increased from an average of (52.4 +/- 9.7)% pre-operation to (85.2 +/- 10.6)% after operation; and the average kyphosis correction of Cobb angle was 11.2 degrees. During an average of 12.6 months follow-up, no patients reported vertebral pain. Review of X-ray films showed no significant loss of vertebral height. One patient died from complications of heart disease 5 months being discharged from hospital. Another patient died from the same cause at 6 months after opertaion. Paravertebral leakage of bone cement was also found in 2 patients. No serious complications occurred and patients were satisfied with the treatment.</p><p><b>CONCLUSION</b>PVP, PKP can be used to instantly reinforce vertebral injury, relieve pain, and improve the patient's mobility. For elder people with fresh thoracolumbar fractures, when health conditions are allowed, PVP or PKP surgery is the right treatment.</p>

Detection of aml-1/eto fusion gene in patients with acute myeloid leukemia by real-time quantitative RT-PCR / 中国实验血液学杂志

This study was aimed to construct the standard product for detecting the aml1/eto fusion gene by real-time quantitative reverse transcription polymerase chain reaction (RQ-RT-PCR), monitoring minimal residual disease (MRD) in the patents with AML-M(2). Having obtained the aml1/eto fusion gene from the patients by the RT-PCR with a pair of specific primers, the RNA standard product as 10(10) copies was gained after amplifying and transcribing in vitro and was used for detecting bone marrow and peripheral blood samples from 37 patients. The results indicated that the standard product constructed above displayed a standard curve which showed a linear correlation of the Ct with the log of the RNA concentration of each standard dilution. The average relative levels of aml1/eto fusion gene in the patients at diagnosis and the patients in relapse were higher than those in patients ongoing complete remission (CR) (p < 0.05). The relative level of aml1/eto fusion gene of the follow-up patients was higher at diagnosis, and lower in patients ongoing CR, then went up again at relapse. The patients whose relative level of aml1/eto fusion gene in CR decreased by 2 log even lower than at diagnosis had a lower risk of relapse. If the relative level of aml1/eto fusion gene kept increasing, the patients had a poor prognosis. It is concluded that RQ-RT-PCR is a sensitive, specific, reliable and reproducible method for monitoring aml1/eto fusion gene. Application of RQ-RT-PCR to detect aml1/eto fusion gene for monitoring MRD in AML-M(2) is helpful to assess the response of therapy and estimate the risk of relapse, RQ-RT-PCR may become an important method to decide the time for intensified therapy and prolong CR for patients.