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Ommaya reservoir implantation is generally used in the treatment of hydrocephalus and intraventricular drug administration. Ommaya reservoir implantation in the subarachnoid space of the spinal cord for the intrathecal drug administration has not been carried out in China, and only several reports can be retrieved from PubMed. About 60%-90% of untreated patients with spinal muscular atrophy type 2 (SMA2) who survive to adulthood often have complex scoliosis and joint deformities. Nusinersen is an effective drug for the treatment of SMA2. And the route of administration is intrathecal injection, which is difficult for patients with severe scoliosis. This article summarizes the process of Ommaya reservoir implantation and postoperative drug administration in a patient with complex scoliosis type SMA2, which provides a new method for clinical treatment of this disease.
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Objective:To study the metabolites of 17β-estradiol via human cytochrome enzyme CYP 1B1 and analyze the genera-tion rate of products by a high performance liquid chromatography coupled with electrochemical detector (HPLC-ECD) method.Meth-ods:A Mightysil RP-18GP (250 mm ×3.0 mm, 5 μm) column was used at the temperature of 40 ℃.The electrochemical detector with E=+900 mV was applied, the mobile phase was 0.5％NaH2PO4(pH 3.0) and methanol (45:55), the flow rate was 0.5 ml · min-1 , and the injection volume was 5μl.Results:The main metabolite was 4-OH-E2 accompanied with a little of 2-OH-E2.The average hydroxylation rate of 4-OH-E2 was about five times as much as that of 2-OH-E2 at the same concentration of estrogen E 2 me-tabolized via CYP1B1.Conclusion:Taken together, CYP1B1 catalyzed hydroxylation sites of 17-beta estradiol based on NADPH me-tabolism are maily 4.
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Objective To optimize the extraction methods of mitochondrial genome DNA(mtDNA)of Oncomelania hupen-sis. Methods The pyrolysis,protein K variable-temperature digestion and high-concentration potassium acetate purification were applied to optimize the high-concentration-salt precipitation method,and then the optimized method was compared with two common extraction methods,the sucrose density gradient centrifugation method and traditional high-concentration-salt pre-cipitation method. The mtDNA samples were identified by using spectrophotometry,agarose gel electrophoresis and the amplifi-cation products of COX1. The nuclear DNA contamination was tested by the amplification products of ITS. Results The concen-tration and yield of the improved method was significantly higher than those of the traditional method(F=3032.65,10185.00, both P<0.01). The mtDNA samples extracted were essentially free of nuclear DNA and protein,meeting PCR,sequence analy-sis and other molecular biology research requirements. Conclusions The improved high-concentration-salt precipitation meth-od for isolating mtDNA is simple,and it has high yield and low cost. The extracted mtDNA can meet relevant analysis require-ments.
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Objective To compare the effects of three kinds of Oncomelania hupensis RNA extraction methods,namely a modified SDS method,TRIzol reagent method,and CTAB method,so as to obtain an economical and efficient method for RNA extraction from O. hupensis. Methods The modified SDS method,TRIzol reagent method and CTAB method were applied to ex-tract the RNA from O. hupensis. A nucleic acid protein analyzer was used to measure the concentration and purity of RNA. The yields were calculated by the concentration of the products. The purity was indicated by A260/A280 and A260/A230. The quality of RNA was inspected by 1% agarose gel electrophoresis. The β-acting gene was selected as the target gene for RT-PCR analysis. Re-sults The RNA yields obtained by using the three kinds of extraction methods were significantly different(F = 16895.85,P <0.01)according to the analysis of variance. The LSD test showed that the yields obtained by using the modified SDS method were the highest,and those obtained by the CTAB method were the lowest. The purity of RNA extracted by the CTAB method was su-perior to that by the other two methods,and the A260/A280 and A260/A230 ratios of the CTAB method were in the range from 1.8-2.0 and 2.0-2.2. The A260/A230 ratios of the other two methods were both lower than 2.0. The RNA extracted by the modified SDS meth-od had the better integrity. The electrophoresis results showed that the 28S rRNA band,18S rRNA band and 5S rRNA band were clear,and there was no obvious smear between each band. The RNA obtained by the TRIzol reagent method had no 28S rRNA band,and that obtained by the CTAB method had no 28S rRNA and 5S rRNA bands. The β-acting gene of the RNA ex-tracted by all the three methods could be amplified by RT-PCR. The costs and time-consuming of the modified SDS method were less than those of the other two methods. Conclusion The modified SDS method is an economic and efficient method,and it is suitable for extracting the RNA of O. hupensis,especially for large sample preparation.
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Objective A new method for detecting K-ras mutations based liquid chip was used to evaluate K-ras mutations associated with non-small cell lung cancer (NSCLC) patients,to direct the personalized treatment and prognosis evaluation.Methods Take the diagnosis technology research methods,the sensitivity and repeatability of the liquid chip K-ras gene mutation detection method were assessed.A total of 100 NSCLC patients from Nov 2011 to Feb 2012 in Shanghai Chest hospital were included in this study,the fresh tumor tissues were collected for DNA extraction.The 2nd exon 12 and 13 codons,containing 8 K-ras mutations occuring in high frequency were amplified by polymerase chain reaction (PCR),followed by ligation of the PCR products to a series of special probes using ligase detection reaction (LDR),then the PCR-LDR products were analyzed by liquid chip platform.Direct sequencing was applied to compare with the detection results.Results The sensitivity of liquid chip technology detection was 10％-20％,higher than the traditional sequencing method by 1％.Average CV value was 4％-15％ and showed good repeatability.5 K-ras mutations in 100 patients (5％) were detected using multiplex PCR-LDR combined fluid chip methods,including 3 Glyl2Val and 2 Gly12Asp mutations in exon 2.The 5 K-ras mutations were verified accurately by direct sequencing.Conclusions The novel detection method of K-ras mutations based PCRLDR and fluid chip shows high throughput,high sensitivity,good repeatability and the results are reliable and accurate.This method can be used to accurately identified K-ras mutations for NSCLC patients prior to their targeted therapy with TKIs.
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Objective To study the clinical and pathological manifestations of microsporidian encephalitis.Methods The clinical findings and the brain pathological features of a patient with microsporidian encephalitis hospitalized in 2004 were studied.Results The onset was subacute or chronic. The body temperature was usually normal or below 37.5℃,but it rose when patient's condition deteriorated and coma appeaxed.The patient had hypoimmunity but without human immunodeficiency virus infection. Multifocal lesions in the whole brain,signs of meningeal irritation and infective myelogram were observed. Rheumatoid factor increased in the early stage and indirect bilirubin,proteins in cerebrospinal fluid(CSF), and immunoglobulin IgG,IgA increased in the middle stage.Cytological examination of CSF showed lymphocyte reaction.Blood routine test showed normal eosinophil granulocyte count.The patient was found to have pleurisy,peritonitis and cystitis.Brain magnetic resonance image(MRI)manifested plaque-like isometric T1 weight image and long T2 weight image signal in white matter of bilateral cerebral hemisphere and cerebella where FLAIR sequence showed hyperintensity.No apparent mass was identified.Contrast- enhanced MRI scan showed patchy and ring-like intensification.The neural system impairments were permanent and not improved after treatment.The pathology of brain tissue showed neuronal degeneration, karyopycnosis and Derivasculitis.The infectious agents were observed in the cytoplasm of neurons.Wister rats had muhiple organ inflammatory reaction 2 weeks after intraperitoneal inoculation of the patient's CSF and a large quantity of pathogens were found in the peritoneal lavage fluid.Conclusions The patient was PAS staining method is useful for detecting the pathogen in neurons and the rate can be raised by animal intraperitoneal cultivation
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Objective To explore the classification of intracranial arachroid cysts(IAC) in CT cisternography(CTC) and its clinicalapplication.Methods 22 cases of IAC diagnosed by plain CT underwent CTC exminaton. IACs were classified into noncommnicatingintracranial arachnoid cyst (NCIAC) and commnicating intracranial arachnoid cyst (CIAC) by wheather or not filled with contrast media in cysts on CTC. NCIAC cases were selected and treated with neuroendoscopic fenestration.Results 15 cases of NCIAC were found by CTC examination. All the NCIAC patients had definite neurologic findings. Postoperatively, all the patients were improved or cured. Follow-upplain CT scan of 9 NCIAC cases showed the cysts were decreased markedly in size, most of the space around the cysts were replaced bynormal cerebral tissue.Conclusion (1)CTC is simple ,safe and specific for making a final diagnosis of IAC. IACs can be classified into CIAC and NCIAC by CTC findings.(2)Neurosurgical indication for IAC is NCIAC patients with symptoms.