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In order to identify the Torque Teno virus (TT virus),a PCR-DHPLC assay was performed in this study.Primers specific were selected according to the characteristics of TT virus nucleic acid sequence to conduct PCR,and PCR products assayed by DHPLC.We analyzed the sensitivity,specificity,repeatability of PCR-DHPLC and applied it preliminarily on clinical detection.The specific testing was performed with TTV,HBV,HCV and HEV,no cross reaction were found,and the PCR-DHPLC assays we developed had good specification and nice repeatability.Sensitivity analysis showed that the developed PCR-DHPLC assays could detect 1.0× 101 copy/μL.Then we detected 32 serum samples by this method,real-time PCR and normal PCR at same time.The results showed that 17 TTV positives results could be observed by PCR-DHPLC for 32 samples,it is consistent with real-time PCR test results and 15 positive by normal RT-PCR.PCR-DHPLC assays showed nice specification,sensitivity,repeatability,and could be used in epidemiological investigation.
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Objective To develop an HPLC wavelength switching combined gradient elution method for simultaneous determi?nation of nine components in Kanglixin Jiaonang(KLXJN). Methods The analysis was performed on a Kromasil C18(4.6 mm×250 mm, 5μm)with gradient elution by using the mobile phase of acetonitrile-methanol(1:2)(A)-0.1%phosphoric acid solution(B). The col?umn temperature was maintained at 30℃and the flow rate was 0.9 ml/min. The detection wavelengths were set at 225 nm for costuno?lide(1)and dehydrocostus lactone(2),254 nm for aloe-emodin(3),rhein(4),emodin(5)and physcion(6),and 425 nm for bisde?methoxycurcumin(7),demethoxycurcumin(8)and curcumin(9). Results The calibration curves were linear within the range(μg/ml)of 6.610-132.2(r=0.9999)for 1,7.890-157.8(r=0.9996)for 2,14.07-281.4(r=0.9992)for 3,3.450-69.00(r=0.9997)for 4, 2.670-53.40(r=0.9998)for 5,3.760-75.20(r=0.9999)for 6,5.880-117.6(r=0.9996)for 7,8.490-169.8(r=0.9993)for 8,and 13.91-278.2(r=0.9991)for 9,respectively. The recoveries for 1,2,3,4,5,6,7,8 and 9 were 98.28%(RSD=1.09%),97.76%(RSD=0.80%),99.08%(RSD=1.72%),97.19%(RSD=1.00%),98.45%(RSD=1.24%),96.96%(RSD=1.21%),98.51%(RSD=1.55%), 97.52%(RSD=0.83%),and 100.04%(RSD=0.93%),respectively. Conclusion The established method is accurate,rapid and can be used for the quality control of KLXJN.
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To explore the factors associated with preoperative epileptic seizure and surgical outcome in patients with cerebral gangliolioma [GG]. A total of 31 consecutive patients with pathologically confirmed ganglioglioma and surgically treated from January 2003 to June 2011 in West China Hospital of Sichuan University were retrospectively reviewed. Clinical data, surgical procedure and follow-up information were collected and analyzed. Nineteen patients presented with epileptic seizure, of which 63.2% were males. The mean age at epilepsy surgery and mean seizure duration were 25.6 years and 2.3 years respectively. Factors associated with preoperative epileptic seizure were supratentorial lesion and temporal lobe involvement [p=0.016 and 0.008]. Intraoperative electrocorticography [ECoG] was applied in 8 out of 19 epilepsy patients. Eighteen achieved total tumor excision. After a mean follow up of 2.8 [1.3-6.3] years, 11 [68.8%, 11/16] achieved seizure free [Engel class I]. Early surgery [seizure duration < 3 years] was a significant predictor of favorable seizure outcome [p=0.013]. None of the factors including seizure type, tumor location, neuroimaging characteristics and application of intraoperative ECoG or surgical strategy were found to be significantly associated with postoperative seizure outcome. Postoperative combination of AEDs was unnecessary for seizure control. Ganglioglioma with temporal lobe involvement usually associated with intractable epilepsy. Early surgical resection is strongly suggested to achieve favorable outcome. Intraoperative ECoG is not inevitable and simple lesionectomy is sufficient for satisfactory seizure control. Early accurate diagnosis of ganglioglioma should be established on comprehensive consideration and plays an important role in dealing with these patients
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Objective: To explore the factors associated with preoperative and postoperative epileptic seizure in patients with cavernous malformations [CMs]
Methods: A total of 52 consecutive patients from January 2009 to June 2011 who underwent surgical treatment in West China Hospital of Sichuan University due to CMs and confirmed by histopathology were retrospectively reviewed.Patients were divided into two groups [epilepsy-group and non-epilepsy group] according to clinical presentation. Other clinical data, treatment procedure, and follow-up information were collected. Engel classification was used to evaluate seizure outcome
Results: Low birth weight, temporal lobe involvement and cortical lesion showed significant difference between two groups [p=0.017, 0.003 and 0.025 respectively]. Cortical lesion highly increased risk for preoperative epileptic seizure [OR=10.48; 95% CI 1.61-68.23]. After a mean follow-up of 2.1 years, 77.8% of epileptic patients achieved Engel class I. Temporal lobe involvement, lesion size < 2.5cm and surgery within one year of symptom onset were found associated with better seizure outcome [p=0.016, 0.012 and 0.050]. Temporal lobe involvement significantly decreased the risk for postoperative epileptic seizure [OR=0.038; 95% CI 0.002-0.833]. Application of ECoG made no significant difference to seizure outcome [p=0.430]. Most patients need continuing medication therapy after surgery
Conclusion: Surgical treatment of patient with CMs is satisfactory in most cases and temporal lobe involvement usually predict favourable postoperative seizure outcome whether under the monitoring of ECoG or not. Thus, epileptic patients with CMs should be considered for surgical treatment especially when cortical brain layer or temporal lobe was involved
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In this study, we report one case of a three-year-old boy infected with Mycoplasma pneumonia (MP) and presenting concomitant multiple organ damage of the heart, kidney, lung and liver, among others, together with a brief review for the diagnosis and treatment of MP infection with multiple organ dysfunction syndrome (MODS).
Subject(s)
Humans , Male , Child , Multiple Organ Failure , Mycoplasma pneumoniae/isolation & purification , Pneumonia, Mycoplasma/pathology , Diagnostic Techniques and Procedures , Infections , PatientsABSTRACT
The cDNA encoding human Vascular Endothelial Growth Factor 165 (VEGF165) was amplified using RT-PCR from human tonsil tissue and cloned into eukaryotic expression vector pcDNA3.1 (+). The recombinant plasmid pcDNA/V was transferred into 293 cells mediated by liposome and the cells stably expressing VEGF were selected under the pressure of G418. ELISA and Western blotting demonstrated that the eukaryotic expression vector pcDNA/V was successfully constructed and its corresponding protein could be expressed efficiently in vitro. Chick Charioallantoic Membrane (CAM) bioassay showed that recombinant protein has biological activity of hVEGF. Model rats with acute myocardial ischemia were used to further study the expression of VEGFin vivo. The model rats were divided randomly into three groups: control group, pcDNA3.1 (+) group and pcDNA/V group. 50microL naked plasmid DNA or saline was intramyocardially injected at three sites into the border zone of infarction. The hearts of rats were excised and fixed histologically, then the infarction sizes were studied by immunohistochemical staining and electron microscope after four weeks. Immunohistochemical staining for VEGF appeared to be negative in control and pcDNA3.1 (+) groups. In pcDNA/V group, myocardial cells in infarction border zone showed positive staining for VEGF in cytoplasm. Ultrastructural anaylsis showed that there were visible hyperplasia of vascular endothilium in pcDNA/V group. The control and pcDNA3.1 (+) groups showed less capillary hyperplasia. In this study, VEGF165 gene was successfully cloned and its protein expressed in vitro and in vivo was of bioactivity, which provides a basis for the further study of biological functions of human VEGF.