Objective To develop a method for rapid and accurate detection and identification of bacterial pathogens directly from body fluid specimens and to evaluate its application in clinical laboratory.Methods Bacteria DNA was extracted from 205 body fluid specimens with column-based kit,and the high variable V1 and V3 regions of bacterial 16S rRNA gene were amplified with broad-range primers.Amplicons were analyzed by pyrosequencing and the generated sequences were searched in the bacterial identification database.Traditional culture-biochemical method was also used for these specimens and the results were taken as the golden standard.SPSS 11.0 was used to calculate the sensitivity,specificity,false positive/negative rate,positive/negative predictive value and positive/negative likelihood rate of pyrosequencing method.Results The positive rate of bacteria culture was 39.5％ (81/205),among which 71 were infected with single bacterium,and 10 were infected with two species of bacteria.Compared with the culture identification results,pyrosequencing had a 100.0％ (71/71) concordance when applied to detect and identify bacterial pathogens from specimens with single specie bacterium infected.To specimens with two species bacteria infected,7 out of 10 specimens were in concordance with the culture identification results.Besides,pyrosequencing detected 10 positive specimens and identified bacterial pathogens infected in the 124 culture-negative specimens.Taken bacteria culture as the standard method,the sensitivity of pyrosequencing for identifying bacterial pathogen in body fluid was 100.0％,and with a specificity of 91.9％,the false positive rate was 8.1％,the false negative rate was 0.0％,the positive predictive value was 89.0％,the negative predictive value was 100.0％,and the positive and the negative likelihood rate were 12.4 and 0,respectively.Conclusion Pyrosequencing can be used to detect and identify bacterial pathogens directly from body fluid specimens with the advantages of rapidity,high sensitivity,high accuracy and high throughput.

Carbon Dioxide ; Dental Enamel ; injuries ; Dental Pulp ; injuries ; Dentin ; injuries ; Erbium ; Hot Temperature ; Humans ; Laser Therapy ; adverse effects ; Lasers ; Lasers, Solid-State ; Neodymium ; Tooth ; radiation effects

Objective To discuss the initial expression of serum procalcitonin (PCT) in patients with severe traumatic brain injury (TBI) and determine the potential value of PCT to predict the neurological outcome.Methods A retrospective analysis was made on patients admitted due to severe TBI (GCS≤8 points) from July 2011 to August 2012.Mortality and neurological outcome of the survivors were determined using Glasgow outcome scale (GOS) at 6 months after TBI.Results A total of 52 patients (39 males and 13 females),at median age of 38 years (range,15-65 years) were included in the study.Twenty-eight patients had good outcome (GOS of grade Ⅳ-Ⅴ),whereas 24 patients had poor outcome or died (GOS of grade Ⅰ-Ⅲ).Within 24 hours after TBI,serum PCT level was significantly higher in patients with bad outcome compared to those with good outcome (0.778 ng/ml:0.094 ng/ml,P ＜0.01).Enhanced PCT level presented a close correlation with the poor outcome (r =0.657,P ＜0.01).Area under the receiver operating characteristic curve (ROC) was 0.879 [95％ CI (0.757,1.000)].A cutoff value of 0.2 ng/ml had a sensitivity of 100％ and a specificity of 72.2％.Once the PCT level was superior to 4.7 ng/ml,none of the patients regained consciousness.Conclusion PCT is a simple and effective method for prediction of the outcome after severe TBI.

Objective To evaluate the clinical value of echocardiography in the diagnosis of aorticoleft ventricular tunnel.Methods The echocardiographic and clinical characteristics were retrospectively analyzed in 6 patients with aortico-left ventricular tunnel we reported and 147 cases that documents reported.Results Six patients were confirmed by operation.The main echocardiographic characteristics of aortico-left ventricular tunnel were as follows:An abnormal communication began in the ascending aorta above the level of the coronary arteries,bypassed the aortic valve,and terminated in the left ventricle.The color Doppler flow imaging demonstrated to-and-fro flow,with or without aortic valve involvement.According Hovaguimian and his colleagues,2 cases were type Ⅰ,2 cases were type Ⅲ,1 case was type Ⅳ,1 case was type Ⅱ (showed as recanalization).Base on all of the 153 patients that we and documents reported,the accuracy rate of ultrasound diagnosis was 79.6％,the misdiagnosis rate was 17.1％,and missed diagnosis rate was 3.3 ％.Conclusions Echocardiography is the non-invasive and sensitive tool in the diagnosis of the type of aortico-left ventricular tunnel and structure of cardiac involvement,and is the preferred method for preoperative.

Objective To evaluate the feasibility, accuracy and its clinical value of MRI-guided needle biopsy of lung lesions. Methods A total of 137 patients with pulmonary nodules or masses underwent lung biopsy in low-field open MRI equipped with iPath 200 optical tracking systems. Among them, 103 cases had solitary pulmonary lesion; the other 34 cases had multiple foci. The maximum diameter of the lesion was not smaller than 3.5 cm ( ≥ 3.5 cm) in 57 patients, between 1.5 cm and 3.4 cm( 1.5-3.4 cm) in 71 patients, not greater than 1.4 cm ( ≤ 1.4 cm) in 9 patients. Results The puncture success rate was 100.0% (57/57) for lesions ≥3.5 cm, 98.6% (70/71) for lesions 1.5-3.4 cm,77.8% (7/9) for lesions ≤1.4 cm and 97. 8% (134/137) for total cases, respectively. According to the pathological results, pulmonary lesions were malignant in 98 cases and benign in 39 cases. The sensitivity,specificity, accuracy, positive predictive value and negative predictive value of MRI-guided lung biopsy were 94.2%(98/104), 100.0% (33/33), 95.6% (131/137), 100.0% (98/98) and 84.6% (33/39),respectively. Conclusion MRI-guided needle biopsy of lung lesion can be performed precisely in a lowfield open MRI with a low risk of complications. As a supplement to US or CT-guided biopsy, it is worth further promotion and application.

Objective To analyze the clinical results and prognostic factors of patients with early-stage primary gastric mucosa-associated lymphoid tissue(MALT) lymphoma. Methods Seventy-seven pa-tients with primary gastric MALT lymphoma treated from 1985 to 2006 were retrospectively analyzed. All pa-tients were pathologically confirmed as MALT lymphoma in stage Ⅰ ,Ⅱ and ⅡE (by modified Blackedge staging system). Thirty-seven patients had stage Ⅰ disease,23 stage Ⅱ and 17 stage ⅡE. Sixty patients un-derwent surgical resection and 17 received non-surgical treatment. Survival rates were calculated by the Kap-lan-Meier analysis with the Logrank test. Results With a median follow up of 57 months for the surviving patients(ranging from 1 to 198 months for all patients), the 5-year overall survival rate, disease-free survival rate,loco-regional control rate and distant metastasis free survival rate were 74% ,70% ,76% and 87% ,re-spectively. In univariate analysis, clinical stage was significantly associated with overall survival. Patients with stage Ⅰ or Ⅱ disease had a better overall survival than those with stage ⅡE (P = 0.01). Tumor size and surgical resection were significantly associated with disease-free survival. Patients with primary tumor 8 cm or less in diameter had better disease-free survival than those with primary tumor more than 8 cm in diameter(P =0.03). Patients who underwent complete resection had better disease-free survival than those who under-went incomplete resection or no surgery (P =0.02). Clinical stage, tumor size and surgical resection were significantly associated with loco-regional control. Patients with stage Ⅰ or Ⅱ disease had better loco-regional control than those with stage ⅡE (P = 0. 03). Patients with primary tumor 8 cm or less in diameter had better loco-regional control than those with primary tumor more than 8 cm in diameter(P =0.01). Patients who un-derwent complete resection had better loco-regional control than those who underwent incomplete resection or no surgery(P=0.03). Patients with stage Ⅰ and Ⅱ disease treated with surgery had more local recurrence, and patients treated without surgery tended to recur systematically. Patients with stage ⅡE disease tended to recur locally in spite of surgery or not. Conclusions The efficacy of surgical and non-surgical treatment for primary gastric MALT lymphoma are similar. Surgical resection is no longer a necessary approach in the primary treatment. Clinical stage is an important prognostic factor for primary gastric MALT lymphoma.

Objective To explore the value of MR imaging-guided percutaneous lumbar discectomy and discolysis with oxygen-ozone mixture for treatment of posterolateral lumbar disc herniation via a new puncture approach of facet joint medial route. Methods All 114 lumbar intervertebral discs in 103 patients were diagnosed as posterolateral lumber disc herniation by CT or MRI, which were located at the levels of L3-4 in 5 cases, LA-5 in 87 cases and L5-S1 in 22 cases. The procedure was guided under 0. 23 T open magnetic resonance with iPath 200 optical tracking system. A 14 G MR-compatible needle was punctured into the disc center via a new puncture approach of facet joint medial route. The therapy steps were as follows: firstly, cut nucleus pulposus and inject 6 ml oxygen-ozone mixture of 60 μg/ml in the disc center;secondly, retreat the needle to the local prominence, cut prominent part and inject 6 ml oxygen-ozone mixture of 60 μg/ml. Thirdly, retreat the needle to the periradicular nerve root, inject 15 ml oxygen-ozone mixture of 40 μg/ml and 4 ml pain-block liquid. All patients were followed up at 3 days, 1 month, 3 months and 6 months after operation, evaluated for the effect of treatment with the modified Macnab criteria, and the results were compared with the χ2 test. Results All procedures were successfully performed. Intraoperative dural injury occurred in 5 cases. Postoperative infection of intervertebral space occurred in 2 cases. The clinical effective rate was 96. 1% (99/103), 84.5% (87/103), 94.2% (97/103), 95.1% (98/103)respectively at 3 days, 1 month, 3 months and 6 months after operation, and the differences were signifieant (χ2 = 12. 942, P = 0. 005 ) . Conclusion MR imaging-guided percutaneous lumbar discectomy and discolysis with oxygen-ozone mixture via facet joint medial route is a minimally invasive, safety and effective method for the treatment of posterolateral lumbar disc herniation.

Objective:To investigate the effect of glycogen synthase kinase-3β inhibitor TWS119 on hunman γδT cells growth and phenotypic characteristics. Methods:Using γδT medium to cultivate human peripheral blood γδT cells in vitro. After co-cultured with different concentrations of glycogen synthase kinase-3β( GSK-3β) inhibitor 4, 6-disubstituted pyrrolopyrimidine ( TWS119 ) for indicated time,growth curve and Wnt/β-catenin activation of in each group were determined by CCK-8 and Western blot assays. The CD62L and CD45RA expression theγδT cells were detected using flow cytometry. Results:Wnt/β-catenin pathway ofγδT cells could activate by TWS119. In the first group,TWS119 could upregulate the expression of CD62L and CD45RA in dose dependent manner while inhibit the growth and ratio of γδT cells. In the second group,TWS119 could promote the growth and ratio of γδT cells with the increase in concentration from 0 μmol/L to 4. 0 μmol/L and decreased thereafter. Besides,TWS119 could promote the expression of CD62L in a dose-dependent and had no effect on the CD45RA. Conclusion: Human γδT cells isolated from peripheral blood treated with TWS119 gave rise to two subsets of CD45RA+CD62L+γδT cells and CD62L+γδT cells.

This study is to investigate the effect of emodin on inducing human myeloid leukemia cell line HL-60 apoptosis and the role of Akt signal pathway in the apoptosis. HL-60 cells were exposed to various dosages of emodin. MTT assay was used to detect HL-60 cell proliferation. Distribution of HL-60 cells in cell cycle was analyzed by flow cytometry and cell apoptosis was observed by MitoCapture apoptosis detection. The protein expressions of Akt signal pathway were detected by Western blotting. The result showed that emodin remarkably inhibited the cell proliferation. The IC50 value for 48 h treatment was about 20 micromol x L(-1). Apoptosis in HL-60 cells could be efficiently induced by emodin in a dose dependent manner and cells were arrested at G0/G1. The expressions of Akt, p-Akt, IkappaB-alpha, p-IkappaB-alpha, p65, p-p65, mTOR and p-mTOR in Akt signal pathway were downregulated after emodin treatment. It can be concluded that emodin could efficiently induce growth inhibition and apoptosis in HL-60 cells. Akt signal pathway may be involved in this process.
Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Emodin ; pharmacology ; HL-60 Cells ; Humans ; I-kappa B Proteins ; metabolism ; NF-KappaB Inhibitor alpha ; Protein Kinases ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; Signal Transduction ; drug effects ; TOR Serine-Threonine Kinases ; Transcription Factor RelA ; metabolism

OBJECTIVETo investigate the correlation between G421C polymorphism in the regulatory region of CYP4F2 gene and essential hypertension and its molecular mechanism.

METHODSTotally 196 hypertensive patients (hypertension group) and 219 normotensive subjects (control group) were genotyped by polymerase chain reaction-restriction fragment length polymorphism. The promoter activity with different alleles was evaluated by reporter assay. A Myb responsive element was identified using gel retardation assay.

RESULTSSignificant differences were found in distribution of genotype and allele frequency of G421C between hypertension group and control group (P < 0.05), and homozygous GG genotype was independently associated with hypertension after adjustment for age, gender, body mass index, and other risk factors (odds ratios 1.87, 95% CI 1.11-3.13, P < 0.05). 421G reporter construct showed decreased promoter activity compared with 421C reporter construct. 421G existed in Myb responsive element, whereas 421C damaged this motif.

CONCLUSIONG421C polymorphism in the regulatory region of CYP4F2 gene is correlated with essential hypertension. 421G allele inhibits transcription by binding affinity of Myb responsive element.

Adult ; Case-Control Studies ; Cytochrome P-450 Enzyme System ; genetics ; Cytochrome P450 Family 4 ; Female ; Gene Frequency ; Genes, myb ; genetics ; Genetic Predisposition to Disease ; genetics ; Genotype ; Humans ; Hypertension ; genetics ; Male ; Middle Aged ; Polymorphism, Genetic ; Regulatory Sequences, Nucleic Acid ; genetics ; Response Elements ; genetics