Study on SNP Genotyping of Degraded DNA by Fluorescence-labeled Multiplex LDR-PCR Amplification / 中国医科大学学报

Objective In this study,a multiplex PCR amplification system was constructed based on fluorescent labeling PCR and LDR,to provide a new strategy for analyzing severely degraded DNA.Methods Eight SNP loci (rs10802248,rs10516197,rs10488372,rs2278945,rs4757318,rs4887255,rs4889002,and rs9304473) were selected.Their LDR probes and PCR primers of linked products were designed and synthesized.Ligase detection reaction,PCR amplification,and capillary gel electrophoresis (CEG) were performed to establish the multiplex LDR-PCR amplification system.Results The genotypes of these 8 loci were obtained simultaneously by the fluorescence-labeled multiplex LDR-PCR amplification method.The loci profiles obtained by fluorescence-labeled multiplex LDR-PCR amplification were in accordance with those obtained by direct sequencing of the polymorphic regions in samples from all individuals.By fluorescence-labeled multiplex LDR-PCR amplification,the 8 SNP loci were efficiently amplified from the severely degraded FFPET DNA.Conclusion Eight SNP loci results could be obtained simultaneously by using the multiplex LDR-PCR amplification system,which is a simple,efficient,and practical SNP genotyping method with accurate and reliable results for highly degraded samples.

Clinicopathologic analysis of primary pulmonary myxoid sarcoma with EWSR1 translocation / 中国癌症杂志

Background and purpose: Primary pulmonary myxoid sarcoma (PPMS) is a very rare lung tumor that has recently been shown to harbor EWSR1-CREB1 translocation. This study aimed to investigate the clinicopatho-logical characteristics and differential diagnosis of PPMS. Methods: The clinical and pathological features of 6 cases of PPMS with EWSR1 gene rearrangement were reviewed. Immunohistochemistry and fluorescence in situ hybridization (FISH) study were performed. Results: Six patients were enrolled in this study, including 4 male patients and 2 female patients with an age range of 23 to 64 years (median age, 44 years). All tumors involved pulmonary parenchyma, with a predominant endobronchial component in 1 and adjacent to bronchus in 5 patients. Microscopically, the tumor was lobulated and composed of cords of polygonal, spindle cells within myxoid stroma. Tumors were immunoreactive for only Vimentin and weakly focal for epithelial membrane antigen (EMA). All tumors were shown to harbor EWSR1 gene rearrangement by FISH. Follow-up of all patients showed that 1 patient developed a pleura and bone metastasis but was still alive and 5 were disease-free after 4-29 months. Conclusion: PPMS is an extremely rare sarcoma with low-grade malignant potential. This is characterized by distinct histological features and EWSR1 gene rearrangement. It is important that understanding tumor spectrum and genetic feature can contribute to diagnosis and differential diagnosis of PPMS.

Effects of light quality on cell growth and psbA promoter of engineered Synechococcus sp. PCC7002 / 生物工程学报

Light quality can regulate both psbA genes and vector promoter psbA of the engineered Synechococcus. Through light regulation, we tried to improve yield of the recombinant protein for vp28 gene-expressed Synechococcus sp. PCC7002. To drive photon-capturing efficiently, three limiting factors (irradiance, temperature and pH) were optimized by measuring net photosynthesis. High cell density cultures were performed with variant ratios of white, red and blue light in a 5-L photo-bioreactor. Yields of biomass, expressions of vp28 and transcription levels of psbA were compared. High ratio blue light-induced vp28 transcription had tripled and the relative accumulation of VP28 protein was doubled. The relative expressions of psbAII and psbAIII had positive correlations with higher ratio of blue light, not the red light. With high ratio red light inducing, dry biomass reached 1.5 g/L in three days. Therefore, we speculated that red light accelerated biomass accumulation of the transgenic strain and blue light promoted transcription for PpsbA and psbA. These results provided useful information for mass production of cyanobacteria and its secondary metabolites.

A comprehensive study on histological features of fusion-positive lung adenocarcinomas and their association with psammoma bodies / 中国癌症杂志

Background and purpose:Gene fusions have been identiifed as recurrent oncogenic events in lung adenocarcinoma. Our purpose are to study the histologic features of anaplastic lymphoma kinase (ALK), c-ros oncogene 1 receptor tyrosine kinase (ROS1) andRETproto-oncogene fusion-positive lung adenocarcinomas and to evaluate the correlation between psammoma bodies and fusion-positive lung adenocarcinomas.Methods:In this study, we performed a comprehensive histologic analysis of 44 fusion-positive (including 15RET, 20ALK and 9ROS1) lung adenocarcinomas and 111 fusion-negative [including 20 epidermal growth factor receptor (EGFR), 20 Kirsten rat sarcoma viral oncogene (K-ras), 71 pan-negative] lung adenocarcinomas.Results:ALK,RET andROS1 fusion-positive lung adenocarcinomas were more prevalent in solid or acinar predominant adenocarcinoma. Multivariate analysis showed that tumors harboring a fusion gene had significantly higher prevalence of the presence of signet ring cells (P=0.000), micropapillary component (P=0.044), mucinous cribriform pattern (P=0.000) and extracellular mucin (P=0.010). The incidence of psammoma bodies was higher in the lung adenocarcinomas with a gene fusion than in tumors without gene fusions (P=0.000). Psammoma bodies were more likely to be found in tumors with any micropapillary component and/or mucinous cribriform pattern than in tumors lacking a micropapillary component and/or mucinous cribriform pattern (P=0.000).Conclusion:Our data showed that the presence of psammoma bodies, micropapillary component, mucinous cribriform pattern, extracellular mucin or signet ring cells may be either sensitive or speciifc to predict tumors harboring a fusion gene. These distinct morphologic features may be helpful in selecting cases for further accurate molecular testing.

Shanghai community-based practice of early lung cancer screening with low-dose spiral computed tomography / 中国癌症杂志

Background and purpose:As one of the most fatal malignant tumors in China, the morbidity and mortality of lung cancer remain high. Early diagnosis and normative treatment is the key to improve the prognosis of lung cancer. The aim of this study was to explore the practice of early lung cancer screening with low-dose spiral computed tomography (CT) based on the current situation in community health service, with integration of superior resources of med-ical institutions at all levels in Shanghai.Methods:From Aug. 2013 to Aug. 2014, we screened high-risk population in selected communities of Minhang District, Shanghai, for early diagnosis of lung cancer with low-dose spiral CT combined with multidisciplinary comprehensive treatment models including minimally invasive surgery, exploring the medical service network covering prevention, diagnosis, treatment, rehabilitation and follow-up.Results:Screening population is 11 332 (male 7 144, female 4 188); Twenty-nine cases with pathological diagnosis of malignant tumor, including 27 cases of pri-mary lung cancer, 1 case of lung metastasis, 1 case of breast cancer. The morbidity of primary lung cancer is 238.26×10-5. There were 22 cases of Stage 0-Ⅰ lung cancer accounting for 81.48% of all diagnosed primary lung cancer.Conclusion:Based on community health service, screening with low-dose spiral CT could improve the early diagnosis rate of lung can-cer with feasibility and validity, which could be applicable in qualified eligible medical center and community in China.

Effect of Low Frequency Ultrasound on Carotid Plaque / 中国康复理论与实践

@#Objective To observe the effects of low frequency ultrasound on carotid artery plaque and artery stenosis. Methods 156 patients with carotid atherosclerosis were divided into treatment group (n=80) and control group (n=76). The control group was administered routine medicine, while the treatment group accepted low frequency ultrasound therapy in addition. The size and shape of carotid artery plaque, severity of stenosis and the level of lipid were observed before and after treatment, and the side-effects were recorded. Results The intima-media thickness (IMT), diameter of plaque, plaque score decreased after treatment in both groups, and decreased more in the treatment group than in the control group (P<0.05); while the frequence of moderate stenosis and severe stenosis was less (P<0.05). The levels of low density lipoprotein- cholesterol and total cholesterol decreased in both groups after treatment (P<0.05), and decreased more in the treatment group than in the control group (P<0.05). No serious side-effect was observed. Conclusion Low frequency ultrasound can reduce the atherosclerotic plaques in carotid artery and relieve the stenosis.

Effect of Low Frequency Ultrasound on Carotid Plaque / 中国康复理论与实践

Objective To observe the effects of low frequency ultrasound on carotid artery plaque and artery stenosis. Methods 156 pa-tients with carotid atherosclerosis were divided into treatment group (n=80) and control group (n=76). The control group was administered routine medicine, while the treatment group accepted low frequency ultrasound therapy in addition. The size and shape of carotid artery plaque, severity of stenosis and the level of lipid were observed before and after treatment, and the side-effects were recorded. Results The intima-media thickness (IMT), diameter of plaque, plaque score decreased after treatment in both groups, and decreased more in the treat-ment group than in the control group (P<0.05);while the frequence of moderate stenosis and severe stenosis was less (P<0.05). The levels of low density lipoprotein-cholesterol and total cholesterol decreased in both groups after treatment (P<0.05), and decreased more in the treatment group than in the control group (P<0.05). No serious side-effect was observed. Conclusion Low frequency ultrasound can reduce the atherosclerotic plaques in carotid artery and relieve the stenosis.

Correlation between diffusion tensor imaging and cognitive dysfunction in patients with delayed encephalopathy after acute carbon monoxide poisoning / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To explore the diagnostic value of magnetic resonance (MR) diffusion tensor imaging (DTI) in detecting brain white matter (WM) damage of patients with delayed encephalopathy after acute carbon monoxide poisoning (DEACMP) and evaluating their cognitive dysfunction.</p><p><b>METHODS</b>Thirteen patients with DEACMP and thirteen age- and sex-matched volunteers underwent DTI using 1.5T MR scanner. FA and ADC values of 16 WM regions of interests (ROIs) were measured on DTI by two experienced radiologists independently with double blind methods, cognitive functions were evaluated by another experienced neurologist blinded to patient's medical history using the Montreal cognitive assessment (MoCA). ADC and FA values in DEACMP patients, and their correlations with cognitive dysfunction were analyzed.</p><p><b>RESULTS</b>ADC values of DEACMP patients increased significantly in all ROIs (P < 0.05) in comparison with the corresponding ROIs of healthy controls, whereas FA values were significantly decreased in all ROIs (P < 0.05) in comparison with that in controls except the bilateral optic radiations, anterior and posterior internal capsules. MoCA scores were positively correlated with FA values of bilateral lower frontal (r(L) = 0.736, P = 0.011; r(R) = 0.762, P = 0.003) lobe, temporal lobe (r(L) = 0.605, P = 0.016; r(R) = 0.559, P = 0.021) and total average WM (r(A) = 0.688, P = 0.001), however it inversely correlated with ADC values of bilateral lower frontal WM (r(L) = -0.674, P = 0.007; r(R) = -0.681, P = 0.019).</p><p><b>CONCLUSION</b>DTI can quantitatively reveal WM microstructure damage of DEACMP patients, indicate the severity of cognitive dysfunctions, and provide important information for pathogenesis and pathological study for DEACMP.</p>

Lentivirus-based RNAi and its application in the study of T lymphocytes / 中国病理生理杂志

The technique of lentivirus-based RNAi ( RNA interference) is a combination of the effective infec-tion and integrating characteristics of lentivirus vector and the specificity of RNAi to silence a homologous gene expression. It caused the post-transcriptional gene silencing ( PTGS) by a specific target sequence of mRNA degradation.As a kind of commonly used target cells in gene therapy, T lymphocytes play an important role in graft-versus-host disease ( GVHD) and other diseases.It is of great significance for increasing the infection efficiency of the target gene to T lymphocytes on this kind of disease for gene therapy.In this paper, we summarize the application of lentivirus-based RNAi in T lymphocytes.

Clinical, pathological and molecular genetic studies on a pedigree with late-onset Pompe's disease complicated with cerebral vascular diseases / 中华神经科杂志

Objective To report a pedigree with late-onset Pompe' s disease complicated with cerebral vascular diseases as to summarize their clinical,pathological and molecular genetic characteristics.Methods We investigated the clinical and pathological data of the two affected siblings with late-onset Pompe' s disease complicated with cerebral vascular diseases.All the 5 members of this pedigree accepted the GAA gene analysis.ResultsBoth affected siblings had progressive pelvic girdle muscle weakness from young adult age,and recently developed vertigo and ataxia.Brain imaging of them revealed multiple cerebral hemorrhage,infarction and diffuse ischemic white matter lesions.The brother had multiple aneurysms and stenoses of cerebral arteries revealed by brain CTA.However,his sister was only found to have multi-beaded stenoses of cerebral arteries.The muscle pathology of the brother showed typical vacuolar degeneration and glycogen storage in muscle fibers. The GAA enzyme activity of 2 siblings were dramatically lower than normal.A heterozygous 19 bp-deletion (c.1388-c.1406,exon 9) were found in GAA gene in the 2 siblings and their healthy mother. Conclusions Cerebrovascular involvement should be a special phenotype of Pompe' s disease.A novel heterozygous mutation c.1388del19 in GAA gene was found in this pedigree,but the relationship between the mutation and the rare clinical phenotype needs further study.

Botulinum Toxin A for Post-stroke Spasm under Color Doppler Ultrasound / 中国康复理论与实践

@#Objective To evaluate outcome of injection of botulinum toxin A to treat post-stroke muscle spasm under color doppler ultrasound guide. Methods 15 stroke patients following spasm in upper limb were chosen. Under the guidance of ultrasound, botulinum toxin A was accurately injected in biceps, brachialis, deep flexor muscle of fingers, superficial flexor muscle of fingers, musculus flexor carpi ulnarista, thumb-muscle, palmaris longus muscle, musculus pronator teresta, and quadrate pronator muscle. All patients went on rehabilitation training after injection. Upper limb function was evaluated by using modified Ashworth(MAS) and Fugl-Meyer scores before injenction, two weeks and four weeks after injection. Results Recovery of muscle spasm and scores of the upper limb motor function were better than that before injection.Conclusion Under color doppler ultrasound guidance, injectionof the botulinum toxin A can treat post-stroke muscle spasm with accurate location.

Expression of calcium-sensing receptor in the rat cardiac tissue at different age and its relation with anoxia-reperfusion injury / 中国病理生理杂志

AIM: To investigate the changes of expression of calcium-sensing receptor(CaSR) in the rat cardiac tissue at different age and its relation with anoxia-reperfusion(A/R) injury.METHODS: RT-PCR was used to detect the mRNA expression of CaSR.The A/R injury was remodeled in vitro,and the ultrastructure of cardiac tissue was observed under transmission electron microscope.RESULTS: The expression of CaSR mRNA in the rat cardiac tissue increased gradually after birth,and reached to its maximum in the 1st month.In the 2nd to 3rd month,the expression were almost equal to that at birth and then increased again to a higher level.The expression level of CaSR mRNA increased more significantly than those in control groups after 40 min of anoxia and reperfusion of 1 h and 2 h, and decreased after 3 h and 4 h.Moreover,the longer the reperfusion time lasted,the more serious the changes of ultrastructure were observed.CONCLUSION: Our results demonstrate that the CaSR is expressed in the rat cardiac tissue.The mRNA expression of CaSR changes with age and is involved in the anoxia-reperfusion injury.

The Biological Characteristics of IL-2 Gene Modified Tumor Cells / 中国肿瘤生物治疗杂志

In this experiment, we introduced human interleukin-2(lL-2)gene and Neomycin resistance (Neo~R) gene into a human lung adenocarcinoma cell line GLC, using a retroviral vector PLXSN. The positive cells obtained by selecting in G418 were compared with the wild-type cells. We observed their growth characteristics in vitro and the quantity of IL-2 in the culture supernatants. PCR result demonstrated the integration of foreign genes into tumor genome DNA after transfection and permanent existence. Under electron microscope, what can be seen was the surface villi of trans-duced-cells are less and shorter. It deserves further investigation to determine whether the change of cell surface influence metastatic ability of cells.

Expression of calcium sensing receptor during ischemia/reperfusion myocardial damage and relationship between CaSR and injury of myocardium / 中国病理生理杂志

AIM: To investigate the expression of calcium sensing receptor(CaSR) during myocardial injury induced by ischemia/reperfusion and disclose the relationship between CaSR and myocardial ischemia/reperfusion. METHODS: The experimental model was established by the 30 min ligating and 1 h, 2 h, 4 h, 6 h reperfusing the left descending coronary artery (LAD) in rats. Wistar rats were randomly divided into 5 groups: sham group, ischemia/reperfusion 1 h, 2 h, 4 h, 6 h groups (I/R 1 h, 2 h, 4 h, 6 h group). CaSR mRNA expression was detected by RT-PCR. Left ventricular function was recorded. The levels of plasma lactate dehydrogenase (LDH), alondialdehyde (MDA) and superoxide dismutase (SOD) were measured. The change of ultrastructure in the ischemia/reperfusion myocardium of rats was observed by electron microscopy. RESULTS: LVSP,?dp/dtmax and SOD activity decreased gradually with the reperfusion time prolonged. LDH and MDA peaked at 2 h. The ultramicro-structural injury at the 1 h and 2 h was more serious than that at 4 h and 6 h. The expression of CaSR increased significantly after reperfusion of 1 h and 2 h, and decreased after 4 h and 6 h. CONCLUSION: The increased expression of CaSR mRNA and serious injure of myocardium were observed. CaSR may be associated with the myocardial ischemia/reperfusion injury.

Effects of spermine on myocardial ischemia/ reperfusion injury in rats / 中国病理生理杂志

AIM: To observe the effect of exogenous spermine (low concentration) on myocardial ischemia/reperfusion injury in rats.METHODS: 40 Wistar rats were randomly divided into 4 groups: sham- operation group (Sham), ischemic reperfusion group (I/R), spermine group (Sp) and natural saline group (NS). The model of ischemic/reperfusion injury was established by ligating rat coronary artery. In Sp group, spermine (0.5 mmol/L, 2 mL/kg) was injected slowly into rat vein. During the process, we recorded the electrocardiogram and the LV functional parameters, assayed the levels of SOD, LDH, NO and MDA in serum, and examined the ultrastructure of the myocardium. RESULTS: In I/R group, the incidence of arrhythmia was 90%, myocardial ultrastructure was injured seriously, values of LVSP and ?dp/dtmax decreased, levels of LDH, NO and MDA increased while SOD activity decreased (P