Objective To investigate the relationship between methylenetetrahydrofotate reductase (MTHFR)C677T polymorphisms and H-type hypertension and increased plasma homocysteine (Hcy) levels. Methods From September 2013 to June 2014,4 012 permanent residents aged ≤30 year from 12 natural villages or communities in 6 regions of Hunan province were extracted according to the cluster random sampling method. Using computer random number table,571 residents were randomly selected as the research objects. According to the blood pressure and Hcy levels,571 residents were divided into 3 groups:a common hypertension group (n = 190),an H-type hypertension group (n = 94),and a normal blood pressure group (n = 287 ). Amplification refractory mutation system-polymerase chain reaction (ARMS-PCR)method was used to detect the MTHFR C677T polymorphisms in all the research objects and the penotyping was performed. Hcy levels were detected at the same time. Results There were significant differences in recessive model (CC + CT,TT)genotype frequencies among the H-type hypertension group (n = 66[70. 2%],n = 28[29. 8%]),common hypertension group (n = 156[82. 1%],n = 34[17. 9%]), and normal blood pressure group (n = 235[81. 9%],n = 52[18. 1%])(χ2 = 6. 797,P = 0. 033),and there were no significant differences in CC,CT,and TT genotype frequencies among the 3 groups (P >0. 05). In the recessive model,there were significant differences in TT genotype frequencies between the H-type hypertension group and the normal blood pressure group or the common hypertension group (χ2 = 5. 812,P = 0. 016;χ2 = 5. 212,P = 0. 022). There was no significant difference in TT genotype frequencies between the common hypertension group and the normal blood pressure group (P > 0. 05). The CC + CT and TT genotype Hcy levels of the MTHFR C677T recessive model in the H-type hypertension group were 17. 1 ±1. 6 and 19. 0 ±2. 9 μmol/ L respectively. There was significant difference between the genotypes (t = - 3. 115,P = 0. 004). The logistic regression analysis of MTHFR C677T recessive model genotype showed that after adjusting for sex and age,the residents with recessive model TT genotype had higher risk of H-type hypertension (OR,1. 946,95% CI 1. 172 -3. 232,P = 0. 01). Conclusion The TT MTHFR C677T gene mutation in this population may be an important genetic factor for the increased Hcy levels and the onset of H-type hypertension.

Objective To investigate the effect of Ser473-Akt phosphorylation in the protection of atorvastatin to cerebral ischemia-re-perfusion (I/R) injury in rats. Methods Forty male Sprague-Dawley rats were randomly divided into normal group (n=10), sham group (n=10), I/R group (n=10) and intervention group (n=10). A model of cerebral ischemia-reperfusion in rats was establishied, with ischemia for 2 hours and reperfusion for 72 hours. The normal group and the sham group received no injury. I/R group was administered with normal saline only, and the intervention group received atorvastatin 10 mg/kg prepared with normal saline at palinesthesia, 24 and 48 hours after reperfu-sion. All rats were sacrificed 72 hours after reperfusion. HE staining and TUNEL staining were performed in the brain specimens. The ex-pression of Akt and Ser473-Akt in the prefrontal cortex of the brain were detected with Western blotting. Results Compared with I/R group, 72 hours after reperfusion, the damage of nerve cells significantly lessened in the intervention group;the apoptosis positive cells significant-ly reduced in the intervention group (t=-6.014, P<0.001). The expression of Ser473-Akt in prefrontal cortex was higher in I/R group than in the normal group and the sham group (t>20.327, P<0.001), and was higher in the intervention group than in I/R group (t=3.649, P=0.007). Conclusion The Ser473-Akt phosphorylation plays an important role in the protection of atorvastatin in nerve cell through anti-apoptosis of nerve cells, and reducing cerebral I/R injury.

Objective To investigate the correlation of T cell subgroups and natural killer (NK) cell's activity level of peripheral blood of the patients with refractory lymphoma. Methods Flow cytometry was applied to detect T cell subgroups' level and NK cell's activity of peripheral blood in 60 early cure lymphoma patients with chemotherapy before and 20 normal controls , after chemotherapy follow-up they were divided into 30 cases of difficult cure group and 30 cases of effective group. Results Compared with the normal controls, CD+4, CD+4/CD+8 and NK cell in lymphoma patients with chemotherapy before decreased (30.17±8.63 vs 46.52±1.39, t =12.218, P ＜0.05; 0.86±0.45 vs 1.64±0.05, t =11.225, P ＜0.05; 12.39±7.08 vs 19.29±0.84,t =6.365, P＜0.05), while CD+3 and CD+8 cell increased (76.14±10.71 vs 70.48±1.44, t =-3.439, P＜0.05;40.28±14.03 vs 28.35±0.73, t =-5.625, P ＜0.05). Compared with effective group, CD+4 CD+4/CD+8 and NK cell in difficult response group with chemotherapy before decreased (27.70±7.81 vs 33.13±8.82, t =2.163, P =0.036;0.67±0.27 vs 1.10±0.52, t =3.272, P =0.003; 9.87±6.60 vs 15.40±6.58, t =2.771, P =0.008), while CD+3 and CD+8 cell increased (79.67±8.18 vs 71.91±12.00, t =-2.540, P =0.015; 44.70±13.99 vs 34.98±12.41, t =-2.416,P =0.020). Conclusion The detection of T cell subgroups' level and NK cell' s activity in early lymphoma patients before chemotherapy may play a role to diagnose and predict the outcome of refractory lymphoma patients.

To make a universal gene targeting vector fitting for most gene and delete positive selection gene after targeting successfully, a vector named pA2T was constructed by inserting one neomycin gene (neo) for positive selection and two same herpes simplex virus thymidine kinase gene HSV-tk1 and HSV-tk2 for negative selection into the vector of pGEM-3Z, and two locus of crossing-over (x) in P1 (LoxP) and two different multiple cloning sites (MCS) were inserted into two flanks of neo separately. There were eight rare cloning sites between neo and HSV-tk1 and five rare cloning sites between neo and HSV-tk2, and neo, HSV-tk1 and HSV-tk2 could be translated respectively in the pA2T. Transfection of the pA2T into goat fetus fibroblast cells with Lipofectamine 2000 conferred resistance to geneticin (G418) and resistance to ganciclovir (GAC) in the cells, which suggested the positive and negative selectable markers could express in the cells and thus the vector pA2T could be used as a universal gene targeting vector. Transformation of the pA2T into the BM25.8 expressing Cre recombinase conferred neo was deleted in the pA2T, which suggested the LoxP was active. Thus, this vector can be inserted by most gene sequences as homologous sequences and positive selection gene can be deleted after targeting successfully, which is very convenience for the production of transgenic animals using gene targeting method.
Animals ; Animals, Genetically Modified ; genetics ; Cloning, Molecular ; Ganciclovir ; pharmacology ; Gene Targeting ; methods ; Genetic Vectors ; genetics ; Gentamicins ; pharmacology ; Goats ; Integrases ; genetics ; Neomycin ; pharmacology ; Phosphotransferases ; genetics ; metabolism