Study on Polymorphisms in the Blood Protein of Tibetan Mastiff

[ Objective] The aim was to study the protein polymorphism in the blood of Tibetan Mastiff, and provide some theoretical basis for resource protection and reasonable development and utilization of Tibetan Mastiff varieties. [ Method] A total of 103 blood samples were taken from four populations of Hequ Tibetan Mastiff, Qinhai Tibetan Mastiff, Tibetan Spaniel and native dogs of Qinghai. Seven blood protein Iocus（Tf, Po, Sα2, Hb, AIb, Pr and Amy）were investigated by using vertical polyacrylamide gel electrophoresis with discontinuous buffer system. Then the genetic variation during different populations was analyzed. [ Result] Genetic variations were observed in Tf, Sα2 and Po in four populations, others were not polymorphic. There were three alleles at the locus of Tf and Po, two alleles at the loci of Sα2. Effective number of alleles and Nei＇s average expected heterozygosity were 1. 532 4 and 0.230 3 relatively, all higher in Tibetan Mastiff than other populations. [ Conclusion] Protein locus in blood of Tibetan Mastiff existed in genetic variation.

Blood diagnostic and prognostic biomarkers in amyotrophic lateral sclerosis

Amyotrophic lateral sclerosis is a devastating neurodegenerative disease for which the current treatment approaches remain severely limited.The principal pathological alterations of the disease include the selective degeneration of motor neurons in the brain,brainstem,and spinal cord,as well as abnormal protein deposition in the cytoplasm of neurons and glial cells.The biological markers under extensive scrutiny are predominantly located in the cerebrospinal fluid,blood,and even urine.Among these biomarke rs,neurofilament proteins and glial fibrillary acidic protein most accurately reflect the pathologic changes in the central nervous system,while creatinine and creatine kinase mainly indicate pathological alterations in the peripheral nerves and muscles.Neurofilament light chain levels serve as an indicator of neuronal axonal injury that remain stable throughout disease progression and are a promising diagnostic and prognostic biomarker with high specificity and sensitivity.However,there are challenges in using neurofilament light chain to diffe rentiate amyotrophic lateral sclerosis from other central nervous system diseases with axonal injury.Glial fibrillary acidic protein predominantly reflects the degree of neuronal demyelination and is linked to non-motor symptoms of amyotrophic lateral sclerosis such as cognitive impairment,oxygen saturation,and the glomerular filtration rate.TAR DNA-binding protein 43,a pathological protein associated with amyotrophic lateral sclerosis,is emerging as a promising biomarker,particularly with advancements in exosome-related research.Evidence is currently lacking for the value of creatinine and creatine kinase as diagnostic markers;however,they show potential in predicting disease prognosis.Despite the vigorous progress made in the identification of amyotrophic lateral sclerosis biomarkers in recent years,the quest for definitive diagnostic and prognostic biomarke rs remains a formidable challenge.This review summarizes the latest research achievements concerning blood biomarkers in amyotrophic lateral sclerosis that can provide a more direct basis for the differential diagnosis and prognostic assessment of the disease beyond a reliance on clinical manifestations and electromyography findings.

Advances in the relationship between apolipoprotein gene polymorphism and blood lipid and cardiovascular disease

Objective： Dyslipidemia is a leading cause of cardiovascular disease. At present, studies have shown that theincidence of cardiovascular disease in our country increased year by year. According to WHO statistics in 2013, itshowed that about 17 million people worldwide die from coronary heart disease （CHD） every year. Currently, CHDis the first cause of death in western countries and the incidence of CHD also showed a trend of increasing. Inrecent years more experts and scholars at home and abroad found gene polymorphism is closely related tohigh-density lipoprotein cholesterol （HDLC） gene and triglyceride （TG） levels. Apolipoprotein （APO） gene is akind of popular polymorphic proteins, whose genetic polymorphisms is through the impact of lipid metabolism,and then closely related to cerebrovascular diseases. But the results are different in different populations and races,or even the opposite. Methods： This review will summarize the gene polymorphism loci of commonapolipoprotein-ApoA1, ApoA5, Apo B, ApoC3, ApoE, which is associated with lipid levels and cardiovasculardisease. Conclusion： It is important for us to get a further understand and prevent the occurrence and developmentof cardiovascular disease from gene level..

Effect of Wheat Based Diet Supplemented with Xylanase on Blood Sugar and Total Protein in Serum of Geese

[Objective] The effects of wheat based diet supplemented with xylanase on blood sugar and total protein in serum of geese were studied. [ Method ] By using the randomized design of single factor, the 1-day-old healthy goslings were divided into 6 groups and fed with corn based diet, wheat based diet and wheat based diet supplemented with xylanase at different concentrations respectively, the contents of blood sugar and total protein in serum were determined. [ Result] The wheat based diet supplemented with xylanase could increase the blood sugar and total protein in serum of geese and wheat based diet supplemented with 0.2% xylanase generated the best effect, which was higher than those of corn based diet group. As for the concentration of protein in serum, wheat based diet supplemented with 0.2% xylanase was significantly different from corn based diet and wheat based diet. [ Conclusion] The wheat based diet supplemented with xylanase could enhance geese production.

TLR2 and TLR4 polymorphisms influence m RNA and protein expression in colorectal cancer

AIM: To evaluate the effect of promoter region polymorphisms of toll-like receptor(TLR)2-196 to-174 del and TLR4-1607T/C(rs10759932) on m RNA and protein expression in tumor tissue and of TLR4+896A/G(rs4986790) on colorectal cancer(CRC) risk.METHODS: The TLR2-196 to-174 del polymorphism was investigated using allele-specific polymerase chain reaction(PCR) and the TLR4-1607T/C and TLR4+896A/G by PCR-restriction fragment length p o l y m o r p h i s m( R F L P). W e g e n o t y p e d 4 3 4 D N A samples from 194 CRC patients and 240 healthy individuals. The m RNA relative quantification(RQ) was performed in 40 tumor tissue samples by quantitative PCR Taq Man assay, using specific probes for TLR2 and TLR4 genes, and ACTB and GAPDH reference geneswere used as endogenous controls. Protein expression was analyzed by immunohistochemistry with specific primary antibodies.RESULTS: No association was found for TLR4-1607T/C and TLR4+896A/G by three statistical models(logadditive, dominant and recessive). However, based on dominant and log-additive models, the polymorphic variant TLR2-196 to-174 del was associated with increased CRC risk [dominant: odds ratio(OR) = 1.72, 95%CI: 1.03-2.89; P = 0.038 and log-additive: OR =1.59, 95%CI: 1.02-2.48; P = 0.039]. TLR2 m RNA expression was increased in tumor tissue(RQ = 2.36) when compared to adjacent normal tissue(RQ = 1; P < 0.0001), whereas the TLR4 m RNA showed a basal expression(RQ = 0.74 vs RQ = 1, P = 0.452). Immunohistochemistry analysis of TLR2 and TLR4 protein expression was concordant with the findings of m RNA expression. In addition, the TLR2-196 to-174 del variant carriers showed m RNA relative expression 2.19 times higher than wild-genotype carriers. The TLR2 protein expression was also higher for the TLR2-196 to-174 del variant carriers [117 ± 10 arbitrary unit(a.u.) vs 95 ± 4 a.u., P = 0.03]. However, for the TLR4-1607T/C polymorphism no significant difference was found for both m RNA(P = 0.56) and protein expression(P = 0.26).CONCLUSION: Our findings suggest that TLR2-196 to-174 del polymorphism increases TLR2 m RNA expression and is associated with higher CRC risk, indicating an important role in CRC genetic susceptibility.

Apolipoprotein E polymorphisms increase the risk of post-stroke depression

Recent reports have shown that apolipoprotein E （APOE） polymorphisms are involved in neurodegenerative disease. However, it is unclear whether APOE affects post-stroke depression. Accordingly, we hypothesized that APOE polymorphisms modify the risk of post-stroke depression. Here, we performed a hospital-based case-control study （including 76 cerebral infarction cases with post-stroke depression, 88 cerebral infarction cases without post-stroke depression, and 109 controls without any evidence of post-stroke depression or cerebral infarction） to determine possible association between APOE rs429358 and rs7412 polymorphisms and risk of post-stroke depression. Our findings show no difference among the groups with regards genotype distribution of the rs7412 polymorphism. In contrast, APOE genotypes with rs429358-C alleles increased the risk of post-stroke depression. Further, the rs429358 polymorphism was associated with significantly decreased regional cerebral blood flow values in the left temporal lobe of post-stroke depression cases. Additionally, the rs429358 polymorphism was not only associated with depression severity, but with increasing serum levels of total cholesterol. These resuits suggest that the APOE rs429358 polymorphism is associated with increased risk of developing post-stroke depression, and that APOE rs429358-C allele genotypes may be detrimental to recovery of nerve function after stoke. Indeed, these findings provide clinical data for future post-stroke depression gene interventions.

Surfactant Protein B 1580 Polymorphism Is Associated with Susceptibility to Chronic Obstructive Pulmonary Disease in Chinese Han Population

Summary: Whether surfactant protein B (SP-B)-18A/C and 1580C/T polymorphism were associated with susceptibility to chronic obstructive pulmonary disease (COPD) in Chinese Han population was investigated. After genomic DNA was isolated from blood of COPD smokers and control smokers, the genotypes of SP-B-18A/C and SP-B1580C/T polymorphism loci were determined by polymerase chain reaction-restriction fragment length polymorphism analysis (PCR-RFLP) respectively. The results showed that there was significant difference in genotypes distribution frequency of SP-B1580C/T polymorphism locus between COPD smokers and control smokers. C→T mutation rate (including TT homozygote and CT heterozygote) in COPD smokers was higher than in control smokers (57.9 % vs 41.7 %, χ2=4.93, P<0.05), whereas there was no significant difference in genotypes distribution frequency of SP-B1580-18A/C locus between COPD smokers and control smokers. The allele frequency (29.1 %) of SP-B1580-18A/C locus is lower than T allele (70.9 %) in Chinese Han Population, and the distribution was different from that in Mexican, in which, the A and T allele frequencies were 85 % and 15 % respectively. It was concluded that SP-B1580 T allele was probably associated with increased susceptibility to COPD in Chinese Han population; The polymorphism of SP-B-18A/C locus maybe varied with race.

Long-term effects of early antibiotic intervention on blood parameters,apparent nutrient digestibility, and fecal microbial fermentation profile in pigs with different dietary protein levels

Backgroud： This study aimed to determine the effects of early antibiotic intervention（EAI） on subsequent blood parameters, apparent nutrient digestibility, and fecal fermentation profile in pigs with different dietary crude protein（CP） levels. Eighteen litters of piglets（total 212） were randomly allocated to 2 groups and were fed a creep feed diet with or without in-feed antibiotics（olaquindox, oxytetracycline calcium and kitasamycin） from postnatal d 7 to d 42. On d 42, the piglets within the control or antibiotic group were mixed, respectively, and then further randomly assigned to a normal-（20%, 18%, and 14% CP from d 42 to d 77, d 77 to d 120, and d 120 to d 185,respectively） or a low-CP diet（16%, 14%, and 10% CP from d 42 to d 77, d 77 to d 120, and d 120 to d 185,respectively）, generating 4 groups. On d 77（short-term） and d 185（long-term）, serum and fecal samples were obtained for blood parameters, microbial composition and microbial metabolism analysis.Results： EAI increased（P 〈 0.05） albumin and glucose concentrations in low-CP diet on d 77, and increased（P 〈 0.05） urea concentration in normal-CP diet. On d 185, EAI increased（P 〈 0.05） globulin concentration in normal-CP diets, but decreased glucose concentration. For nutrient digestibility, EAI increased（P 〈 0.05）digestibility of CP on d 77. For fecal microbiota, the EAI as well as low-CP diet decreased（P 〈 0.05） E. coli count on d 77. For fecal metabolites, on d 77, EAI decreased（P 〈 0.05） total amines concentration but increased skatole concentration in low-CP diet. On d 185, the EAI increased（P 〈 0.05） putrescine and total amines concentrations in low-CP diets but reduced（P 〈 0.05） in the normal-CP diets. The low-CP diet decreased the concentrations of these compounds.Conclusions： Collectively, these results indicate that EAI has short-term effects on the blood parameters and fecal microbial fermentation profile. The effects of EAI varied between CP levels, which was characterized by the significant alteration of glucose and putrescine concentration.

Association of Surfactant Protein B Gene Polymorphisms (C/A-18, C/T1580, Intron 4 and A/G9306) and Haplotypes with Bronchopulmonary Dysplasia in Chinese Han Population

Summary： This study aimed to investigate the association between surfactant protein B （SP-B） pol- ymorphisms and bronchopulmonary dysplasia （BPD） in Chinese Han infants. We performed a case- control study including 86 infants with BPD and 156 matched controls. Genotyping was performed by sequence specific primer-polymerase chain reaction （PCR） and haplotypes were reconstructed by the fastPHASE software. The results showed that significant differences were detected in the geno- type distribution of C/A-18 and intron 4 polymorphisms of SP-B gene between cases and controls. No significant differences were detected in fhe genotype distribution of C/T1580 or A/G9306 be- tween the two groups. Haplotype analysis revealed that the frequency of A-del-C-A haplotype was higher in case group （0.12 to 0.05, P=0.003）, whereas the frequency of C-inv-C-A haplotype was higher in control group （0.19 to 0.05, P=0.000）. In addition, a significant difference was observed in the frequency of C-inv-T-A haplotype between the two groups. It was concluded that the polymor- phisms of SP-B intron 4 and C/A-18 could be associated with BPD in Chinese Han infants, and the del allele of intron 4 and A allele of C/A-18 might be used as markers of susceptibility in the disease. Haplotype analysis indicated that the gene-gene interactions would play an important part in deter- mining susceptibility to BPD.

Association between Low-density Lipoprotein Receptor-related Protein 5 Polymorphisms and Type 2 Diabetes Mellitus in Han Chinese:a Case-control Study

Objective To investigate the association between low-density lipoprotein receptor-related protein 5 （LRPS） variants （rs12363572 and rs4930588） and type 2 diabetes mellitus （T2DM） in Han Chinese. Methods A total of 1842 T2DM cases （507 newly diagnosed cases and 1335 previously diagnosed cases） and 7777 controls were included in this case-control study. PCR-RFLP was conducted to detect the genotype of the two single nucleotide polymorphisms （SNPs）. Odds ratios （ORs） and 95% confidence intervals （95% CIs） were calculated to describe the strength of the association by logistic regression. Results In the study subjects, neither rs12363572 nor rs4930588 was significantly associated with T2DM, even after adjusting for relevant covariates. When stratified by body mass index （BMI）, the two SNPs were also not associated with T2DM. Among the 3 common haplotypes, only haplotype ~ was associated with reduced risk of T2DM （OR 0.820, 95% CI 0.732-0.919）. In addition, rs12363572 was associated with BMI （P〈0.001） and rs4930588 was associated with triglyceride levels （P=0.043） in 507 newly diagnosed T2DM cases but not in healthy controls. Conclusion No LRP5 variant was found to be associated with T2DM in Han Chinese, but haplotype TT was found to be associated with T2DM.

Relationship between nerve injury-induced protein gene 2 polymorphism and stroke in Chinese Han population

The aim of present study was to investigate the relationship between nerve injury-induced protein 2 （NINJ2） gene polymorphism and stroke in Chinese Han population. Fifty-two patients with large-artery atherosclerosis （LAA） infarction, 85 patients with small-artery occlusion lacunar （SAO） infarction, 50 patients with intracerebral hemorrhage （ICH） and 66 controls were included. Genotypes and alleles frequencies of the two single nucleotide polymorphisms （SNPs） of NINJ2 among different groups were analyzed and compared. In regard to rs12425791, the frequencies of the AG and AA＋AG genotypes of the LAA and SAO groups were significantly higher than those in the control group; the frequency of the A allele of the SAO group was significantly higher than that of the control group. In regard to rs11833579, there were not any significant differences between the case and the control groups. The SNP rs12425791 is significantly associated with ischemic stroke, and the A allele increases the susceptibility to stroke. The SNP rs11833579 is not significantly associated with stroke.

Productive performance and blood metabolites as affected by protected protein in sheep

This investigation included two experiments. Experiment 1 was executed to study the effect of feeding different rations of protected protein of canola meal on digestibility and nutritive values within sheep. Twenty male, healthy sheep were divided into five treatments according to the methods of protein protection (control, heat, sodium hydroxide, formaldehyde, and acetic acid treatments). Experiment 2 was carried out on developing lambs to investigate the effect of protected protein on growth performance and some blood metabolites. Animals in this ex-periment were also divided into the same treatments as Experiment 1. Animals in the first and second experiment were fed concentrate ration (80%) and wheat straw (20%) to cover the feed requirements. Nutritive values expressed as total digestible nutrients (TDN %) and digestible crude protein (DCP%) of the experimental rations was calculated. In the second experimental all animals were weighed biweekly and the amounts of rations were adjusted throughout the experimental period (120 days) according to their body weight change. Results indicated that in the first experimental protected protein by heat (HE) and sodium hydroxide (NH) had positive (P<0.05) effects on most of digestibility coefficients of different nutrients. Protein protection methods also improved (P<0.05) the nutritive values (TDN and DCP) in the HE treatment and NH treatment. In the second experiment body weight increased by 14% and 7% and also daily gain by 27% and14 % in HE and NH, respectively, while FM and AC decreased body weight by 8% and 4.4%. Higher values (P<0.01) in both thyroid hormones were observed in HE and NH than those other treatments. Also, higher values (P<0.01) of total protein, albumin, and glucose were observed in HE and NH than other treatments. The control (CTL) group recorded higher concentrations of urea-N and creatinine at different periods of the experiment in comparison with other treatments. Generally, from the present investigation it can be concluded that protected protein of canola meal by heat or sodium hydroxide treatments were more efficient for productive performance and some blood metabolites of sheep.

Surfactant Protein A Polymorphism Is Associated with Susceptibility to Chronic Obstructive Pulmonary Disease (COPD) in Chinese Uighur Population

This study investigatedexamined the correlation between surfactant protein-A (SP-A) polymorphism and the susceptibility of cvhronic obstructive pulmonary disease (COPD) in Xinjiang Uighurs. Genomic DNA was extracted from peripheral blood of 194 COPD smokers and 201 healthy smokers of Uighur who were hospitalized in or paid a visit to one of the four Xingjiang-based hospi-tals involved in the study, betweenfrom March 2009 to December 2010. Single nucleotide polymor-phisms (SNPs) were studied on A/G atwithin amino acid aa62 (CCA/CCG rs1136451) and C/T within aa219 (CGG/TGG, rs4253527) in SP-A. Genotypes were determined by using the TaqMan polymerase chain reaction (PCR). Our results showed that genotype frequencies were different be-tween the COPD and normal smokers infor aa62 (χx2=6.852, P=0.033). There were also significant differences in allele genotype frequencies between the COPD and the control and allele G might de-crease the risk COPD (χx2=6.545, P=0.011; OR=0.663; 95% CI: 0.484–0.909). The result suggested We were led to conclude that polymorphism of aa62 (CCA/CCG, rs1136451) of SP-A may be asso-ciated with the susceptibility to COPD in Xingjiang Uighurs.

Genetic Polymorphism of Milk Protein and Their Relationships with Milking Performances in Chinese Yak

The milk protein polymorphisms were typed by polyacrylamide gel electrophoresis (PAGE)from 109 Maiwa and 100 Jiulong yaks, and the relationships among milk protein polymorphisms,milking traits and milk protein compositions were studied. The results showed thatβ-CN,κ-CN andα-La were monomorphic,αs1-CN andβ-Lg were polymorphic, the dominantgenes were αs1-CN D and β-Lg E,respectively. The frequencies of αs1-CN D were 0.8073and 0.6000 and β-Lg E were 0.9770 and 0.9700 in two populations respectively.The meanheterozygosities were 0.1021 and 0.1867 in two populations. No significant effects onmilking traits and milk protein compositions were observed except for αs1-CN locus onfat percentage in Jiulong yak.

G-protein beta 3 subunit polymorphisms and essential hypertension： a case-control association study in northern Han Chinese

Objective To explore the association between the three polymorphisms [ C825T, C1429T and G（-350）A] of the gene encoding the G protein beta 3 subunit （GNB3） and hypertension by performing a case-control study in the northern Han Chinese population. Methods We recnaited 731 hypertensive patients and 673 control subjects （the calculated power value was 〉 0.8）. Genotyping was performed to identify C825T, C1429T and G（-350）A polymorphisms using the TaqMan assay. Comparisons of allelic and genotypic frequencies between cases and controls were made by using the chi-square test. Logistic regression analyses were performed to investigate the relationships between the three polymorphisms of GNB3 gene under different genetic models （additive, dominant and recessive models）. Results The genotype dis- tribution and allele frequencies of C825T, C1429T and G（-350）A polymorphisms did not differ significantly between hypertensive patients and control subjects, either when the full sample was assessed, or when the sample was stratified by gender. No significant association was observed between C825T, C 1429T and G（-350）A polymorphisms and the risk of essential hypertension in any genetic model. Linkage dis- equilibrium was only detected between C825T and C 1429T polymorphisms. Haplotype analyses observed that none of the three estimated haplotypes significantly increased the risk of hypertension. Conclusions Our study suggested that the GNB3 gene polymorphisms [C825T, C 1429T and G（-350）A] were not significantly associated with essential hypertension in northern Han Chinese population.

Correlation between intakes of carbohydrates,protein,and fat with random blood sugar levels in menopausal women

Objective: Carbohydrates, proteins, and fats are energy sources needed by the body for performing daily activities and generating primary energy substances. In women who have undergone menopause, the function of thyroid hormone in their body begins to decline and thus affects the ability of the body to produce energy. The purpose of this study was to determine the correlation between the intakes of carbohydrates, proteins, and fats and random blood sugar levels in menopausal women.Methods: This study was a correlational analytical research with a cross-sectional design, which was conducted in 72 menopausal women recruited by the purposive sampling technique. In this study, a 24-hour food recall form was used, and randomized blood sugar levels were measured using a glucometer. The analysis of the data was performed using a Pearson product moment and multiple linear regression.Results: Carbohydrates and fats together affected random blood sugar levels with an F-value of 25.810 and a p-value of 0.000.Meanwhile, adjusted R^2 showed the value of 0.411, indicating that the difference in the intake of carbohydrates and fats together affected random blood sugar levels by 41.1%.Conclusions: Intake of carbohydrates and fats affected random blood sugar levels, whereas the rest were influenced by other factors,and protein intake was unrelated to random blood sugar levels in menopausal women.

Polymorphisms of microsomal triglyceride transfer protein in different hepatitis B virus-infected patients

AIM: To identify the two polymorphisms of microsomal triglyceride transfer protein (MTP) gene in the Chinese population and to explore their correlation with both hepatitis B virus (HBV) self-limited infection and persistent infection. METHODS: A total of 316 subjects with self-limited HBV infection and 316 patients with persistent HBV infection (195 subjects without familial history), matched with age and sex, from the Chinese Han population were enrolled in this study. Polymorphisms of MTP at the promoter region -493 and at H297Q were determined by the allele specific polymerase chain reaction (PCR). RESULTS: The ratio of males to females was 2.13:1 for each group and the average age in the self-limited and chronic infection groups was 38.36 and 38.28 years, respectively. None of the allelic distributions deviated significantly from that predicted by the Hardy-Weinberg equilibrium. There was a linkagedisequilibrium between H297Q and -493G/T (D’ = 0.77). As the χ2 test was used, the genotype distribution of MTP -493G/T demonstrated a significant difference between the self-limited infection group and the entire chronic group or the chronic patients with no family history (χ2 = 8.543, P = 0.015 and χ2 = 7.199, P = 0.019). The allele distribution at the MTP-493 position also demonstrated a significant difference between the study groups without family history (χ2 = 6.212, P = 0.013). The T allele emerged as a possible protective factor which may influence the outcomes of HBV infection (OR: 0.59; 95% CI: 0.389-0.897). CONCLUSION: The polymorphism of the MTP gene, T allele at -493, may be involved in determining the HBV infection outcomes, of which the mechanism needs to be further investigated.

Expression of multidrug resistance 1 gene and C3435T genetic polymorphism in peripheral blood of patients with intractable epilepsy

BACKGROUND： Increased expression of multidrug resistance 1 （MDR1） mRNA in peripheral blood of patients with intractable epilepsy is not due to epilepsy drugs, but epilepsy behavior. Monitoring MDR1 expression in peripheral blood is a target for MDR1 gene evaluation. OBJECTIVE： To investigate the influence of antiepileptic drugs and seizures on MDR expression in intractable epilepsy, and to analyze the genetic polymorphisms of C3435T in the MDRl gene. DESIGN, TIME AND SETTING： Factorial designs and comparative observations at the experimental center of the Affiliated Hospital of Qingdao Medical College, Qingdao University between October 2003 and October 2004. PARTICIPANTS： A total of 120 subjects were recruited from the epilepsy clinical department of the Affiliated Hospital of Qingdao Medical College. Four groups （n = 30） were classified according to statistical factorial design： intractable epilepsy, treatment response, no treatment, and normal control groups. METHODS： One-step semi-quantitative reverse-transcription polymerase chain reaction technology was used to test expressions of the MDR1 gene in 120 subjects. C3435T polymorphisms in intractable epilepsy group and normal control groups were analyzed by polymerase chain reaction-restriction fragment length polymorphism. MAIN OUTCOME MEASURES： Expression of MDR1 mRNA in the four groups, and C3435T genetic polymorphisms in intractable epilepsy and normal control groups. RESULTS： MDRl gene expression was increased in the intractable epilepsy group, due to the factor seizures, but not the antiepileptic drugs. However, the interaction between the two factors was not statistically significant. Of the 30 subjects in the intractable epilepsy group, the following genotypes were exhibited： 3 （10%） C/C genotype, 9 （30%） C/T genotype, and 18 （60%） T/T genotype at the site of C3435T, while 4 （13%）, 10 （33%）, and 16 （53%） subjects were determined to express these genotypes in the normal control group, respectively. C and T allele frequency were 25% and 75% in the intractable epilepsy group, and 30% and 70% in the normal control group, respectively. However, there was no statistical difference between the groups. CONCLUSION： Results demonstrated that seizures, not antiepileptic drugs, induced MDR1 gene expression in intractable epilepsy. Genetic polymorphisms of C3435T in the MDR1 gene did not contribute to the development of multidrug resistance in patients with intractable epilepsy.

An epidemiologic study of mitochondrial membrane transporter protein gene polymorphism and risk factors for neural tube defects in Shanxi, China

The present study involved a questionnaire survey of 156 mothers that gave birth to children with neural tube defects or had a history of pregnancy resulting in children with neural tube defects （case group） and 156 control mothers with concurrent healthy children （control group） as well as detection of mitochondrial membrane transporter protein gene [uncoupling protein 2 （UCP2）] polymorphism. The maternal UCP2 3＇ untranslated region （UTR） D/D genotype and D allele frequency were significantly higher in the case group compared with the control group （odds ratio （OR） 3.233; 95% confidence interval （C/） 1.103 9.476; P= 0.040; OR： 3.484; 95% CI： for neural tube defects 2.109 5.753; P 〈 0.001）. Univariate and multivariate logistic regression analysis of risk factors for neural tube defects showed that a matemal UCP2 3＇ UTR D/D genotype was negatively interacted with the mothers＇ consumption of frequent fresh fruit and vegetables （S = 0.007）, positively interacted with the mothers＇ frequency of germinated potato consumption （S = 2.15） and positively interacted with the mothers＇ body mass index （S = 3.50）. These findings suggest that maternal UCP2 3＇ UTR gene polymorphism, pregnancy time, consumption of germinated potatoes and body mass index are associated with an increased risk for neural tube defects in children from mothers living in Shanxi province, China. Moreover, there is an apparent gene-environment interaction involved in the development of neural tube defects in offspring.

Association of gene and protein expression and genetic polymorphism of CC chemokine ligand 4 in colorectal cancer

BACKGROUND Leukocytes,such as T cells and macrophages,play an important role in tumorigenesis.CC chemokine ligand(CCL)4,which is produced by lymphocytes and macrophages,has been found to be expressed in the mucosa of the gastrointestinal tract and is a potent chemoattractant for various leukocytes.AIM To examine CCL4 expression and its genetic polymorphism rs10491121 in patients with colorectal cancer(CRC)and evaluate their prognostic significance.METHODS Luminex technology was used to determine CCL4 Levels in CRC tissue(n=98),compared with paired normal tissue,and in plasma from patients with CRC(n=103),compared with healthy controls(n=97).Included patients had undergone surgical resection for primary colorectal adenocarcinomas between 1996 and 2019 at the Department of Surgery,Ryhov County Hospital,Jönköping,Sweden.Reverse transcription quantitative PCR was used to investigate the CCL4 gene expression in CRC tissue(n=101).Paired normal tissue and TaqMan single nucleotide polymorphism assays were used for the CCL4 rs10491121 polymorphism in 610 CRC patients and 409 healthy controls.RESULTS The CCL4 protein and messenger RNA expression levels were higher in CRC tissue than in normal paired tissue(90%,P<0.001 and 45%,P<0.05,respectively).CRC tissue from patients with localized disease had 2.8-fold higher protein expression levels than that from patients with disseminated disease.Low CCL4 protein expression levels in CRC tissue were associated with a 30%lower cancer-specific survival rate in patients(P<0.01).The level of plasma CCL4 was 11%higher in CRC patients than in healthy controls(P<0.05)and was positively correlated(r=0.56,P<0.01)with the CCL4 protein level in CRC tissue.The analysis of CCL4 gene polymorphism rs10491121 showed a difference(P<0.05)between localized disease and disseminated disease in the right colon,with a dominance of allele A in localized disease.Moreover,the rate of the A allele was higher among CRC patients with mucinous cancer than among those with nonmucinous cancer.CONCLUSION The present study indicates that the CRC tissue levels of CCL4 and CCL4 gene polymorphism rs10491121,particularly in the right colon,are associated with clinical outcome in CRC patients.