1.Gene mutation analysis of X-linked hypophosphatemic rickets.
Ying SONG ; Hong-Wei MA ; Fang LI ; Man HU ; Shuang REN ; Ya-Fen YU ; Gui-Jie ZHAO
Chinese Journal of Contemporary Pediatrics 2013;15(11):928-931
OBJECTIVETo investigate the frequency and type of PHEX gene mutations in children with X-linked hypophosphatemic rickets (XLH), the possible presence of mutational hot spots, and the relationship between genotype and clinical phenotype.
METHODSClinical data of 10 children with XLH was retrospectively reviewed. The relationship between gene mutation type and severity of XLH was evaluated.
RESULTSPHEX gene mutations were detected in all 10 children with XLH, including 6 cases of missense mutation, 2 cases of splice site mutation, 1 case of frameshift mutation, and 1 case of nonsense mutation. Two new mutations, c.2048T>C and IVS14+1delAG, were found. The type of PHEX gene mutation was not associated with the degree of short stature and leg deformity (P=0.571 and 0.467), and the mutation site was also not associated with the degree of short stature and leg deformity (P=0.400 and 1.000).
CONCLUSIONSMissense mutation is the most common type of PHEX gene mutation in children with XLH, and c.2048T>C and IVS14+1delAG are two new PHEX gene mutations. The type and site of PHEX gene mutation are not associated with the severity of XLH.
Adolescent ; Child ; Child, Preschool ; Familial Hypophosphatemic Rickets ; genetics ; Female ; Humans ; Infant ; Male ; Mutation ; PHEX Phosphate Regulating Neutral Endopeptidase ; genetics ; Retrospective Studies
2.Evaluation of oxidative stress in colorectal cancer patients.
Dong CHANG ; Fan WANG ; Ya-Shuang ZHAO ; Hong-Zhi PAN
Biomedical and Environmental Sciences 2008;21(4):286-289
OBJECTIVETo evaluate the oxidative stress in patients with colorectal cancer and to investigate the relationship between oxidative stress and colorectal cancer.
METHODSSeventy-six subjects were divided into two groups (36 colorectal cancer patients as the study group and 40 normal healthy individuals as the control group). Their protein oxidation, DNA damage, lipid peroxidation and antioxidants, vitamin C, vitamin E, glutathione (GSH), and antioxidative enzymes in serum were detected.
RESULTSThe levels of protein carbonyl and advanced oxidation protein products (AOPP) were significantly higher in the study group than in the control group (P<0.01). Serum 8-OHdG was significantly increased in the study group compared to the control group (P<0.01). However, the mean serum level of MDA and conjugated diene was lower in the study group than in the control group (P<0.01). The activity of antioxidative enzymes was significantly decreased in the study group compared to the control group (P<0.01). Serum vitamins C and E concentrations were significantly reduced in the study group compared to the control group (P<0.01).
CONCLUSIONColorectal cancer is associated with oxidative stress, and assessment of oxidative stress and given antioxidants is important for the treatment and prevention of colorectal cancer.
Adult ; Aged ; Aged, 80 and over ; Colorectal Neoplasms ; metabolism ; DNA Damage ; Female ; Humans ; Lipid Peroxidation ; Male ; Middle Aged ; Oxidative Stress
3.Expression of recombinant human IFNa-2b/IgG4 Fc fusion protein in a baculovirus insect cell system.
Zhao-xia JI ; Ya-ning CHEN ; Yan-rui ZHANG ; Yu-xiu YANG ; Chun-rong WANG ; Shuang-yin HAN
Chinese Journal of Hepatology 2012;20(8):617-620
To investigate a baculovirus insect cell system for expressing an interferon alpha 2b (IFNa2b)/immunoglobulin G-4 (IgG4) Fc fusion protein, which has long-acting antiviral effects. Human IFNa2b and IgG4 Fc cDNAs were generated by molecular cloning and inserted into a baculovirus shuttle vector, which was then transposed into the DH10 Bac strain to form recombinant Bacmid-IFN/Fc. The Bacmid-IFN/Fc was transfected into High five insect cells, and expression of the IFN/Fc fusion protein was detected by Western blotting and its biological activity was assessed by the cytopathic effect inhibition method. The IFNa2b and IgG4 Fc cDNA fragments were successfully amplified by RT-PCR using human peripheral lymphocytes. After cloning into the baculovirus shuttle vector, pFastBac1, and transforming into DH10 Bac competent cells, screening identified positive clones carrying the recombinant Bacmid-IFN/Fc. A Bacmid-IFN/Fc clone was successfully transfected into the High five insect cells and packaged into the baculovirus for expression of the IFN/Fc fusion protein. Western blotting revealed that the fusion protein expression was specific, and yielded a protein of 45 kD in size. The in vitro antiviral activity of the IFN/Fc fusion protein was 580 IU/mL. A novel IFN/Fc fusion protein was successfully generated using a baculovirus insect cell system, which may prove useful for providing future experimental data for development of a new long-acting interferon to treat chronic viral hepatitis.
Animals
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Antiviral Agents
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metabolism
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Baculoviridae
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genetics
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Cell Line
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Cloning, Molecular
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Gene Expression
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Gene Fusion
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Genetic Vectors
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Humans
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Immunoglobulin Fc Fragments
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biosynthesis
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genetics
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Immunoglobulin G
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biosynthesis
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genetics
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Insecta
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Interferon-alpha
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biosynthesis
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genetics
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Recombinant Fusion Proteins
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biosynthesis
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genetics
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Recombinant Proteins
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biosynthesis
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genetics
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Reverse Transcriptase Polymerase Chain Reaction
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Transfection
4.Effect of (S)-4C3HPG on brain damage in the acute stage of moderate traumatic brain injury model of mice and underlying mechanism.
Nan YANG ; Shuang-Shuang DAI ; Ya-Lei NING ; Xing-Yun CHEN ; Yan ZHAO ; Ping LI ; Yuan-Guo ZHOU
Acta Physiologica Sinica 2010;62(6):555-559
The aim of this study is to investigate the effect of (S)-4-carboxy-3-hydroxy-phenylglycine [(S)-4C3HPG], a mixed group I glutamate metabotropic receptor antagonist and a group II agonist, on impairment in a cortical impact model of traumatic brain injury (TBI) in mice and to elucidate the possible mechanisms. Mice were injected (i.p.) with saline, 1 mg/kg (S)-4C3HPG, 5 mg/kg (S)-4C3HPG and 10 mg/kg (S)-4C3HPG (n=10 per group), respectively, at 30 min before moderate TBI. Neurological deficit scores, water content in injured brain and glutamate concentration in cerebral spinal fluid (CSF) were detected at 24 h after TBI. The expressions of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) mRNA in injured cortex were also detected by real-time RT-PCR. The results showed that the neurological deficits and cerebral edema were significantly attenuated in mice pretreated with (S)-4C3HPG (5 and 10 mg/kg respectively) compared with those in mice pretreated with saline. Furthermore, (S)-4C3HPG treatment also decreased the glutamate concentration in CSF and the expressions of TNF-α and IL-1β mRNA remarkably in a dose-dependent manner. These results suggest that (S)-4C3HPG treatment attenuates cortical impact-induced brain injury possibly via suppression of glutamate release and inhibition of excessive inflammatory cytokine production. These findings highlight the potential benefit of glutamate metabotropic receptor ligand for preventing TBI.
Animals
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Brain Injuries
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drug therapy
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metabolism
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physiopathology
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Cytokines
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metabolism
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Glutamic Acid
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cerebrospinal fluid
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Glycine
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analogs & derivatives
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therapeutic use
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Male
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Mice
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Mice, Inbred C57BL
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Receptors, Metabotropic Glutamate
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agonists
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antagonists & inhibitors
5.Relationship between single nucleotide polymorphisms in the deoxycytidine kinase gene and chemosensitivity of gemcitabine in six pancreatic cancer cell lines.
Shuang SI ; Quan LIAO ; Yu-Pei ZHAO ; Ya HU ; Qiang ZHANG ; Li-Li YOU
Chinese Medical Journal 2011;124(3):419-422
BACKGROUNDSingle nucleotide polymorphisms (SNPs) in the deoxycytidine kinase (dCK) gene are associated with chemosensitivity to nucleoside analogs. 2',2'-Difluoro 2'-deoxycytidine (gemcitabine) is a first-line nucleoside analog drug in the treatment of pancreatic cancer. However, the association between SNPs in the dCK gene and chemosensitivity to gemcitabine has not been fully established. Therefore, the present study aimed to investigate the relationship between SNPs in the dCK gene and chemosensitivity to gemcitabine in human pancreatic cancer cell lines.
METHODSSeven SNPs in the dCK gene were sequenced in six human pancreatic cancer cell lines. The chemosensitivity of these six cell lines to gemcitabine were evaluated in vitro with a Cell Counting Kit-8 (CCK-8) test. Inhibition rates were used to express the chemosensitivity of pancreatic cancer cell lines to gemcitabine.
RESULTSThe genotype of the A9846G SNP in the dCK gene was determined in six human pancreatic cancer cell lines. The cell lines BxPC-3 and T3M4 carried the A9846G SNP genotype AG, whereas cell lines AsPC-1, Mia PaCa2, SW1990 and SU86.86 carried the GG genotype. Cell lines with the AG genotype (BxPC-3 and T3M4) were more sensitive to gemcitabine compared with cell lines with the GG genotype (AsPC-1, Mia PaCa2, SW1990 and SU86.86) and significantly different inhibition rates were observed between cell lines carrying the AG and GG genotypes (P < 0.01).
CONCLUSIONSVariants in the A9846G SNP of the dCK gene were associated with sensitivity to gemcitabine in pancreatic cancer cell lines. The dCK A9846G SNP may act as a genetic marker to predict chemotherapy efficacy of gemcitabine in pancreatic cancer.
Antimetabolites, Antineoplastic ; pharmacology ; Cell Line, Tumor ; Cell Survival ; drug effects ; Deoxycytidine ; analogs & derivatives ; pharmacology ; Deoxycytidine Kinase ; genetics ; Genotype ; Humans ; Pancreatic Neoplasms ; enzymology ; genetics ; Polymorphism, Single Nucleotide ; genetics
6.Awareness on SARS and public health emergencies among general publics.
Shuo WANG ; Bin-you WANG ; Chong PENG ; Cui-ping SONG ; Hong-xia ZHANG ; Dian-jun SUN ; Wei-wei LI ; Ya-shuang ZHAO ; Wenjing TIAN ; Shu-zhen WEI ; Sheng-yuan LIU
Chinese Journal of Epidemiology 2006;27(6):503-507
OBJECTIVETo explore the awareness on sever acute respiratory syndrome (SARS) and public health emergencies among general publics.
METHODSA cluster sampling method was implemented in Harbin and Jiagedaqi district of Daxinanling of Heilongjiang province. Research subjects were divided into three groups as city, township and rural areas and were given questionnaires to fill in. Data was analyzed with Epi-data and SPSS.
RESULTS2003 available questionnaires were collected. The general publics well understood the knowledge on public health emergencies and the SARS with the whole recognition rate more than 60 percent. During the epidemics, people in city, town and countryside were calm (71.7%). The rates of attitude towards the government were significantly different among the subjects living with the city, town or rural areas. The city group expressed the highest favor to the government and media, 71.8% of them gave the credit on the control of SARS to the effective method taken by the government and 65.0% of them showed that they had enough confidence on the governmental ability of dealing with crises while the countryside group trusted the hospitals and relative specialists the most.
CONCLUSIONIt is essential for the government to interact and communicate with the publics through media, medical and related institutions when confronting with the public health emergencies. Publicity on health knowledge and coping system on emergency should play key roles in the development of an effective public health system while the government should lead the battle.
China ; Data Collection ; Emergencies ; Health Knowledge, Attitudes, Practice ; Humans ; Public Health ; Rural Population ; Severe Acute Respiratory Syndrome ; Urban Population
7.Mining the specifically expressed genes in sperms based on the bioinformatics methods.
Chun-qiong FENG ; Ya-guang ZOU ; Tie-qiu LI ; Qi-zhao ZHOU ; Fei LI ; Shuang LIANG ; Xiang-ming MAO
Journal of Southern Medical University 2009;29(2):185-190
OBJECTIVETo analyze the specifically expressed genes in sperms for better understanding of the molecular characteristics of sperms.
METHODSThe hybridization data the genes in the sperms, oocytes and 10 normal tissues were retrieved from the GEO database to identify the genes expressed specifically in sperms and the patterns of their regulation using such bioinformatic tools as GATHER, PANTHER and DAVID.
RESULTS AND CONCLUSIONSComparison of the spermatozoal gene expression profiles with those of the normal tissues identified 8998 differentially expressed probes, among which 25 genes were up-regulated by over 200 folds in the sperms. Comparison of the gene expression profiles between the oocytes and normal tissues resulted in the identification of 8981 differentially expressed probes. Of the 1709 up-regulated genes in the sperm with a ratio>5, 1218 genes showed similar expressions in the oocytes and the normal tissues, and 129 were up-regulated and 362 down-regulated in the oocytes. The 362 genes up-regulated in the sperms but down-regulated in the oocytes were involved mainly in protein modification and metabolism and nucleic acid metabolism, but very few participated in the intracellular signaling pathways. Numerous transcriptional factors containing the KRAB domain and receptor- independent serine/threonine kinase were specifically overexpressed in sperms, and the a very high proportion of the genes specifically overexpressed in the sperms coincided with the overexpressed genes in the neural stem cells and embryonic stem cells. The genes involved in the glycolysis were down-regulated in the sperms. These findings in the genes specifically expressed in the sperms by data mining using bioinformatic methods may provide better insight into the molecular characteristics of the sperms.
Adult ; Computational Biology ; methods ; Data Mining ; Gene Expression Profiling ; methods ; Humans ; Male ; Spermatozoa ; cytology
8.Clinical analysis and genetic diagnosis of short-limb inherited short stature diseases in children.
Fang LI ; Hong-Wei MA ; Ying SONG ; Man HU ; Shuang REN ; Ya-Fen YU ; Gui-Jie ZHAO
Chinese Journal of Contemporary Pediatrics 2013;15(11):932-936
OBJECTIVETo analyze the clinical manifestations, bone X-ray findings and genetic analysis results of three short-limb inherited short stature diseases: achondroplasia (ACH), hypochondroplasia (HCH) and pseudoachondroplasia (PSACH).
METHODSThe clinical manifestations, bone X-ray findings, and genetic analysis results of 10 children with genetically confirmed short-limb inherited short stature diseases, including 4 cases of ACH 3 cases of HCH, and 3 cases of PSACH, were analyzed.
RESULTSThe 10 patients had a mean body height of -3.69±1.79 SD, a mean sitting height/standing height ratio of 0.65±0.03, and a mean finger spacing/body height ratio of 0.93±0.04. Four ACH cases and 3 PSACH cases showed typical bone X-ray findings; one HCH case showed a smaller sciatic notch, and another HCH case showed no widening of interpedicular distance. G380R mutation in FGFR3 gene was detected in 3 of 4 ACH cases, and Y278C mutation in the other ACH case, N540K mutation in FGFR3 gene was detected in 3 HCH cases, and heterozygous mutations in COMP gene were detected in 3 PSACH cases.
CONCLUSIONSChildren with ACH and PSACH have severer short stature and skeletal deformities than children with HCH, who have mild, atypical clinical manifestations. Bone X-ray and genetic analysis are helpful for the diagnosis and differential diagnosis of the three diseases. The mutational hotspots in two genes are involved in the three diseases, which is conducive to clinical genetic diagnosis.
Achondroplasia ; diagnostic imaging ; genetics ; Bone and Bones ; abnormalities ; diagnostic imaging ; Child ; Child, Preschool ; Dwarfism ; diagnostic imaging ; genetics ; Female ; Humans ; Limb Deformities, Congenital ; diagnostic imaging ; genetics ; Lordosis ; diagnostic imaging ; genetics ; Male ; Mutation ; Radiography ; Receptor, Fibroblast Growth Factor, Type 3 ; genetics
9.Association of catalase genotype with oxidative stress in the predication of colorectal cancer: modification by epidemiological factors.
Dong CHANG ; Zhang Liang HU ; Lin ZHANG ; Ya Shuang ZHAO ; Qing Hui MENG ; Qing Bai GUAN ; Jin ZHOU ; Hong Zhi PAN
Biomedical and Environmental Sciences 2012;25(2):156-162
OBJECTIVEThis paper aims to assess the interaction between common variations in catalase (CAT) polymorphic gene and environmental factors for antioxidant defense enzyme in modulating individual susceptibility to colorectal cancer (CRC).
METHODSA case-control study with 880 colorectal cancer cases and 848 controls was conducted to investigate whether variations in the catalase (CAT) gene, one of the genes involved in scavenging oxidative stress, influenced susceptibility to CRC.
RESULTSThe interaction between life style and genotypes as well as with their effects on colorectal cancer was deduced from the present study. Significant difference (P = 0.01) was identified in the distribution of CAT genotype between the colorectal cancer cases and the controls. The CRC cases had significantly lower mean activity than the controls (P < 0.01). Correlation analyses revealed statistically significant correlations between CAT activity and CAT genotype (P < 0.01).
CONCLUSIONThe risk of CRC was associated with smoking, low vegetable consumption, high pork and poultry consumptions, and low or high BMI. This is the first study reporting an association of polymorphism CAT-21A > T with colorectal cancer. Low CAT activity was associated with an increased risk of CRC; however, no evidence was found to support an association between CAT-21A > T polymorphism and CRC risk.
Adult ; Aged ; Case-Control Studies ; Catalase ; genetics ; Colorectal Neoplasms ; enzymology ; epidemiology ; metabolism ; Female ; Genotype ; Humans ; Male ; Middle Aged ; Oxidative Stress ; Polymerase Chain Reaction
10.Analysis on the clinical characteristics and related risk factors of patients with hemorrhagic transformation after cerebral infarction
Jia-Jun CHEN ; Shuang ZHANG ; Yun-Hua ZHAO ; Xi-Ya JIN ; Yuan-Bing LI ; Li-Hong HUANG
Chinese Journal of Epidemiology 2012;33(11):1176-1180
Objective To analyze and summarize the clinical characteristics and risk factors for patients with hemorrhagic transformation (HT) after cerebral infarction to provide guidance for its clinical treatment and prevention.Methods In this study,data from 49 hospitalized patients with HT in the First Department of Neurology,China-Japan Union Hospital of Jilin University from October 2009 to March 2012,were reviewed retrospectively and 106 cases with acute cerebral infarction only during the same period,were chosen randomly as controls.Gender and age of the patients were comparable.Relevant information was collected.SPSS 17.0 statistical package was applied for data processing.Qualitative data were processed with x2 test,and measurable data was processed with t-test.Each index was analyzed with uni-variate analysis while statistically significant risk factors were included in the logistic review model to conduct the multivariate regression analysis.Results (1) Clinical symptoms deteriorating after hemorrhage in 21 cases accounted for 42.9% of the HT group,among which the cases on degree of disturbance to consciousness had an aggravation in 8 cases and hemiplegia increase in another 7 cases.Headaches and dizziness were found in 5 cases.(2)Number of infarction within 15 days after the occurrence of HT accounted for 87.0%.(3)HT-related factors increased the risk of HT in cerebral infarction such as cortical infarction,large area of infarction,atrial fibrillation,cerebral embolism,diabetes and high level of low-density lipoprotein cholesterol (P<0.05).The most important factors were atrial fibrillation and cerebral embolism.(4)PH-2 seemed more unlikely to link with clinical symptoms than other subtypes of HT.Conclusion Cerebral infarction after occlusion of the main artery trunk was prone to HT,especially when it appeared within 15 days.Short-term prognosis seemed to be related to the subtypes of HT,with risk factors as cortical infarct,massive cerebral infarction,atrial fibrillation,cerebral embolism,diabetes,high low-density lipoprotein cholesterol etc.on HT.