2.Dynamic Observation of Attachment and Invasion of Toxoplasma gondii Tachyzoites to Intestinal Mucosa in BALB/c Mice by Chromogenic In Situ Hybridization Targeting SAG2 mRNA
Xiaoming MA ; Xiaoli MENG ; Guorong YIN ; Hongli LIU ; Jinyan SHEN
Chinese Journal of Parasitology and Parasitic Diseases 1987;0(04):-
Objective To observe dynamically the location and time of attachment and invasion of Toxoplasma gondii tachyzoites to murine intestinal mucosa by chromogenic in situ hybridization targeting SAG2 mRNA. Methods Thirty 7- to 8-week-old BALB/c mice were randomly divided into experiment group(24 mice)and control group(6 mice). Each animal in the experiment group was given 2?104 tachyzoites of RH stain in 0.2 ml PBS by intragastric administration and that in the control group was given 0.2 ml PBS. Four mice in the experiment group and one in the control group were sacrificed at 15 min,30 min,1 h,2 h,4 h and 8 h after infection,respectively,and paraffin sections of duodenum,jejunum and ileum were prepared to perform the in situ hybridization with Dig-labeled oligonucleotide probe complementary to SAG2 mRNA of T. gondii. Results Tachyzoites were found on the striated border of small intestine epithelial cells (absorptive cells,goblet cells and endocrine cells),in or between two absorptive cells or in the lamina propria. At 15 min-2 h after infection,there was significant difference in the number of attachment on jejunum and ileum (P
3.Exploration of homogeneous care at Hunan Provincial People's Hospital
Yuelan QIN ; Zeya SHI ; Zhoumin SHEN ; Yimin ZHU ; Xiaomao SHI ; Jinyan ZHOU ; Yi YANG
Chinese Journal of Hospital Administration 2016;32(6):470-472
An introduction to the Basic service module featuring holistic,full flow and seamless connection built by the hospital.Authors described their experiences in achieving homogenous clinical nursing care,and pointed out that integrating medical humanistic care into the daily nursing process,can improve doctor-patient relationship and patients satisfaction,thus raising medical service quality and efficiency.
4.The epidemiological survey of hypertension among the elderly population in Shanxi Province
Yuying HOU ; Shufang ZHAO ; Huaxiang RAO ; Xia LIU ; Ya JING ; Jinyan SHEN ; Huizhen WANG ; Zhishen LIU ; Qinghua ZHANG ; Jianjin GUO
Chinese Journal of Geriatrics 2008;27(8):628-630
Objective To investigate the prevalence rate of hypertension among the elderly population in Shanxi Province. Methods According to the random sampling, 41 residents committees of city blocks and villages of 9 areas (including Taiyuan, Datong, Jinzhong, Yuncheng, et al) of Shanxi Province were extracted as the investigative spots. All the people were older than 55years, and risk factors for hypertension were identified. Results A total of 3702 people were surveyed, including 1,782 men and 1,920 women. The total hypertension prevalence rate was 39.0%(the standardized rate: 39. 5 %), with 38. 7 % in men (the standardized rate: 38. 5%), 39.3 % in women (the standardized rate: 41.2%), and no statistic significance was found between men and women (χ2= 0. 143, P>0. 05). Aging, lacking of knowledge, and obesity were risk factors for hypertension. Conclusions The hypertension in Shanxi Province has a high prevalence rate among the elderly population, a comprehensive intervention should be taken in the prevention of hypertension.
5.Characterization of M2 gene of H3N2 subtype swine influenza virus.
Xiaodu WANG ; Peijun CHEN ; Yang SHEN ; Yafeng QIU ; Xufang DENG ; Zixue SHI ; Lina PENG ; Jinyan LUO ; Chao LIU ; Zhiyong MA
Chinese Journal of Biotechnology 2010;26(1):16-21
M2 protein of influenza A virus is encoded by a spliced mRNA derived from RNA segment 7 and plays an important role in influenza virus replication. It is also a target molecule of anti-virus drugs. We extracted the viral genome RNAs from MDCK cells infected with swine influenza A virus (SIV) H3N2 subtype and amplified the SIV M2 gene by reverse transcriptase-polymerase chain reaction using the isloated viral genome RNAs as template. The amplified cDNA was cloned into a prokaryotic expression vector pET-28a(+) (designated pET-28a(+)-M2) and a eukaryotic expression vector p3xFLAG-CMV-7.1 (designated p3xFLAG-CMV-7.1-M2), respectively. The resulted constructs were confirmed by restriction enzyme digestion and DNA sequencing analysis. We then transformed the plasmid pET-28a(+)-M2 into Escherichia coli BL21 (DE3) strain and expressed it by adding 1 mmol/L of IPTG (isopropyl-beta-D-thiogalactopyranoside). The recombinant M2 protein was purified from the induced bacterial cells using Ni(2+) affinity chromatography. Wistar rats were immunized with the purified M2 protein for producing polyclonal antibodies specific for it. Western blotting analysis and immunofluorescence analysis showed that the produced antibodies were capable of reacting with M2 protein expressed in p3xFLAG-CMV-7.1-M2-transfected cells as well as that synthesized in SIV-infected cells. We also transfected plasmid p3xFLAG-CMV-7.1-M2 into Vero cells and analyzed its subcellular localization by immunofluorescence. The M2 protein expressed in the Vero cells was 20 kDa in size and dominantly localized in the cytoplasm, showing a similar distribution to that in SIV-infected cells. Western blotting analysis of SIV-infected cells suggested that M2 was a late phase protein, which was detectable 12 h post-infection, later than NS1, NP and M1 proteins. It would be a potential molecular indicator of late phases replication of virus. Our results would be useful for studying the biological function of M2 protein in SIV replication.
Animals
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Antibodies, Monoclonal
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biosynthesis
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Cercopithecus aethiops
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Cloning, Molecular
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Escherichia coli
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genetics
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metabolism
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Influenza A Virus, H3N2 Subtype
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genetics
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RNA
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biosynthesis
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genetics
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Rats
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Rats, Wistar
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Recombinant Proteins
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biosynthesis
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genetics
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immunology
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Swine
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Transfection
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Vero Cells
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Viral Matrix Proteins
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biosynthesis
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genetics
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Virus Replication
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genetics
6.High incidence of sudden cardiac death in one family with type-3 long QT syndrome: molecular genetics and electrophysiology mechanism analysis.
Hui ZHOU ; Zhang LI ; Ghani Ali RAZA ; Wengen ZHU ; Qiongqiong ZHOU ; Yang SHEN ; Jinyan XIE ; Qing CAO ; Rong WAN ; Jinzhu HU ; Kui HONG
Chinese Journal of Cardiology 2015;43(12):1046-1050
OBJECTIVESWe identified the long QT syndrome (LQTS) patients, and detected the potential risk of LQTS in family members by using genetic testing and electrophysiological analysis, which helped provide clinical evaluation and appropriate treatment.
METHODSDetailed clinical characteristics and familiar history were obtained from the whole family members of an idiopathic pediatric LQTS patient. Two hundred healthy subjects with the same ethnic background were recruited as controls. The entire coding sequences of three candidate genes including KCNQ1, KCNH2 and SCN5A were screened for mutations in the proband. The function of the mutation was then explored by whole-cell patch clamp techniques, and the genetic testing and risk assessment of the family members were performed.
RESULTSThe proband was clinically preliminary diagnosed as LQTS by 12-lead electrocardiogram. On the third day of metoprolol intake (25 mg, bid), she died suddenly at lunch. One heterozygous missense mutation (SCN5A-V411M) was identified in this proband, but the mutation was absent in 200 healthy subjects. The electrophysiological analysis indicated that SCN5A-V411M significantly increased the peak current density ((230.8 ± 27.6)pA/pF vs. (101.2 ± 10.9)pA/pF, n=10, P<0.01) and the late sodium current ((156.6 ± 13.6)pA/pF vs. (95.9 ± 7.9)pA/pF, n=12, P<0.01) of sodium channel compared to wide type. The enhanced sodium channel activation with a negative shift in the peak I-V relationship was significantly higher by -50 mV than wide type (85.0%± 7.4% vs. 41.5% ± 2.6%, P<0.01), while the steady-state inactivation curves remained unchanged. Additionally, mother and grandmother of the proband were the silent mutation carriers with no symptoms, who needed the appropriate clinical assessment and follow-up. The proband's twin sister and aunt died of sudden infant death syndrome.
CONCLUSIONSWe firstly reported a heterozygote missense mutation (SCN5A-V411M) in this Chinese family. V411M induced "gain of function" of sodium channel and formed the basis of type-3 LQTS. Genetic testing could help to increase the diagnostic accuracy, and facilitate clinical assessment and appropriate therapy to prevent sudden cardiac death of individuals with SCN5A-V411M mutation.
Cardiac Conduction System Disease ; Death, Sudden, Cardiac ; Genetic Testing ; Humans ; Incidence ; Long QT Syndrome ; Mutation ; Patch-Clamp Techniques
7. Recurrent syncope related to catecholaminergic polymorphic ventricular tachycardia due to de novo RyR2-R2401H mutation
Xin LIU ; Juxiang LI ; Jinzhu HU ; Yang SHEN ; Rong WAN ; Qinmei XIONG ; Qiongqiong ZHOU ; Jinyan XIE ; Jiejing JIN ; Xia YAN ; Jianhua YU ; Kui HONG
Chinese Journal of Cardiology 2017;45(1):39-43
Objective:
To explore the clinical and molecular genetic features of a Chinese patient with catecholaminergic polymorphic ventricular tachycardia (CPVT).
Methods:
Clinical data including resting electrocardiography, echocardiography and treadmill exercise testing of a patient with CPVT admitted to our department in March 2013 were analyzed, and the peripheral venous blood samples of the patient and his family members and 400 ethnicity-matched healthy controls were obtained. All exons and exon-intron boundaries of the six CPVT-related genes including RYR2, CASQ2, TRDN, CALM1, KCNJ2 and ANKB were sequenced to detect the variants related to CPVT. The relationship between the genotypes and phenotypes was analyzed to direct the target therapy.
Results:
Recurrent syncope induced either by exercise or extreme frightened fear was observed in this patient. There was no positive family history of syncope or sudden death. The resting electrocardiography and echocardiography of the patient were normal, while the exercise testing revealed bidirectional and polymorphic ventricular tachycardia. A cardiac ryanodine receptor gene mutation (R2401H) was identified in this patient, while this mutation was absent in his parents and sister and 400 controls. No variant was detected in the remaining five candidate genes. Treatment with high dose of metoprolol succinate (118.75 mg/d) was effective and patient was free of syncopal attack during the 2 years follow-up.
Conclusion
This is the first report on RyR2-R2401H mutation in Chinese patient with CPVT, and high dose of metoptolol is the effective therapy option for CPVT related to RyR2 mutation.
8.Effect of kangaroo care on neurological behavior in neonates with neonatal intensive care unit asphyxia
Danqiong LU ; Jinyan WANG ; Xia YU ; Jinya WANG ; Junying SHEN ; Suping WANG
Chinese Journal of Practical Nursing 2020;36(12):909-912
Objective:To explore the effect of kangaroo care on the neurological behavior of neonatal intensive care unit (NICU) neonates with asphyxia.Methods:Totally 76 cases of neonatal asphyxia admitted to NICU from January 2018 to December 2018 were randomly divided into observation group (40 cases) and observation group (36 cases). The control group was given routine nursing care, while the observation group was given kangaroo nursing intervention on the basis of the control group. The neurobehavioral changes, the occurrence of nervous system damage and the physical development after 42 days of birth were compared between the two groups.Results:The scores of behavior ability (11.64 ± 0.26), original reflex (5.89 ± 0.32), passive muscle tension (7.64 ± 0.46), active muscle tension (7.83 ± 0.55), general reaction (6.17 ± 0.46) in the intervention observation group were significantly higher than those in the control group (11.02 ± 0.39), original reflex (5.53 ± 0.31), passive muscle tension (7.21 ± 0.47), active muscle tension (6.17 ± 0.46) Tension (7.41 ± 0.41), general reaction (5.88 ± 0.41) ( t values were 8.227, 4.970, 4.027, 3.740, 2.888, P<0.05). The incidence of neurological damage in the observation group was 2.50% (1/40) which was significantly lower than 16.67% (6/36) in the control group ( χ 2=4.117, P < 0.05). The growth of body weight (2.17 ± 0.42) kg, length (7.15 ± 1.74) cm and head circumference (4.38 ± 0.93) cm in the observation group were significantly higher than those in the control group (1.68 ± 0.39) kg, (5.89 ± 1.81) cm, (3.81 ± 0.79) cm ( t values were 5.252, 3.093, 2.863, P<0.05). Conclusion:Kangaroo nursing can effectively promote the neurological development and physical development of neonatal asphyxia, and reduce the nervous system damage of asphyxiated neonates, and the operation is simple and easy, which is worthy of clinical application.
9.The predictive value of small densed low-density lipoprotein cholesterol and HbA 1C for cardiovascular and cerebrovascular events in elderly patients with type 2 diabetes mellitus
Jingfen LU ; Jinyan ZHAO ; Xu CHEN ; Honglin YANG ; Qiong LI ; Feifei LU ; Hua YOU ; Hao SHEN
Chinese Journal of Laboratory Medicine 2020;43(3):250-254
Objective:To investigate the serum level of low density lipoprotein cholesterol (sdLDL-C) in elderly patients (age≥65 years) with type 2 diabetes mellitus (T2DM) and the its predictive value in evaluatingthe risk of cardiovascular and cerebrovascularevents in elderly patients with T2DM.Methods:In this retrospective study,386 elderly patients with T2DM were collected from December 2014 to December 2016, the averageage was (72.7±5.4) years old, including 269 males and 117 females; 92 of whom had cardiovascular events during follow-up, the averageage was (72.9 ± 5.2) years old, including 65 males and 27 females, and serum sdLDL-C level was detected. Meanwhile, biomarkers such as lipoprotein (a), apolipoprotein B, apolipoprotein AI and hypersensitive c-reactive protein were analyzedin all the patients.Univariate and multivariate factors were used to analyze the relationship between each variable and the occurrence of cardiovascular and cerebrovascular events. The correlation between LDL-C, sdLDL-C of all subjects with age and other lipid indexes were analyzed. ROC curve was used to determine the predictive value of sdLDL-C elevation for cardiovascular and cerebrovascular events in elderly patients with T2DM.Results:The levels of LDL-c, sdLDL-C, non-HDL-C, GLU, HbA1c and ApoB were significantly higher in theCardio-cerebral vascular event group ( t=3.26, 3.46, 2.91, 2.47, 4.03, 3.00, P<0.05). While the levels of apolipoprotein AI was significantly lower than those in theNon-cardio-cerebral vascular event group ( t=-2.39, P<0.05). Cox regression analysis showed that sdLDL-C per 10 mg/dl was independently correlated with the risk of cardiovascular and cerebrovascular events( HR 1.281, 95 %CI 1.225-16.032, P<0.01)after adjusted for age. SdLDL-C was positively correlated with TG, non-HDL-C and ApoB ( r=0.323, 0.588, 0.623, P<0.01) and was negativelycorrelated correlation with age, HDL-C and apolipoprotein AI ( r=-0.363, -0.301, P<0.01), ROC curve analysis showed that sdLDL-C had a strong ability to predict the occurrence of cardiovascular and cerebrovascular events (AUC=0.736, 95 %CI 0.554 9-0.918 2, P=0.003), while HbA1c also had a strong ability to predict the occurrence of cardiovascular and cerebrovascular events(AUC=0.725, 95 %CI 0.524 3-0.927 3, P=0.006). Combined sdLDL-C with HbA1had the strong ability to predict the occurrence of cardiovascular and cerebrovascular events (AUC=0.837, 95 %CI 0.711 4-0.973 5, P=0.001). Conclusions:The elevation of serum sdLDL-C in elderly patients with T2DM wasa significantly independentrisk factorof cardiovascular and cerebrovascular events. Serum sdLDL-C had a higher clinical value than LDL-C, which was expected to be the most effective predictor of lipid profile in riak assessment of cardiovascular and cerebrovascular events in elderly patients with T2DM.