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

Objective To evaluate the clinical value of neutrophil CD64(nCD64)index as a novel biomark-er in the differential diagnosis of acute and chronic brucellosis.Methods A total of 38 patients with acute bru-cellosis and 48 patients with chronic brucellosis diagnosed in the People's Hospital of Xinjiang Uygur Autono-mous Region from February 2021 to July 2023 were included.Peripheral blood of the patients was collected and nCD64 index was detected by flow cytometry,and the correlation between nCD64 index and disease severi-ty was analyzed.Receiver operating characteristic(ROC)curve was used to analyze the sensitivity and the specificity of nCD64 index in differentially diagnosing acute and chronic brucellosis.Meanwhile,Rose-Bengal Plate Test(RBPT)and Standard Tube Agglutination Test(SAT)were used as controls to evaluate the clini-cal diagnostic value of the three.Results The nCD64 index of acute brucellosis patients was higher than that of chronic brucellosis patients(U=216.00,P<0.001),and the index was positively correlated with the sever-ity of the disease(r=0.670,P<0.001).The ROC curve analysis results showed that the area under the curve of nCD64 index in the differential diagnosis of acute and chronic brucellosis was 0.882(95%CI:0.811-0.952,P<0.001),the cut-off value was 2.81,and sensitivity,specificity,positive predictive value,negative predictive value and accuracy were 83.3%,81.6%,80.4%,81.9%and 82.6%,respectively.The efficacy of nCD64 index in differential diagnosis of nCD64 index was significantly better than those of the qualitative tests of RBPT and SAT.Conclusion nCD64 index has favourable sensitivity and specificity in the differential diag-nosis of acute and chronic brucellosis,and tends to reflect the severity of the disease.It has clinical value in the differential diagnosis of acute brucellosis and chronic brucellosis,and plays an important role in the early diag-nosis and treatment effect monitoring of brucellosis.

OBJECTIVE To explore the mechanism of Baihe Dihuang decoction in the treatment of Alzheimer's disease(AD) based on network pharmacology, molecular docking and animal experiment. METHODS TCMSP were used to predict the active components and targets of Baihe Dihuang decoction and disease-related targets were collected from GeneCards, OMIM and DRUGBANK databases, respectively. Target protein interactions were analyzed with STRING database and biological function and pathway were analyzed with Metascape database. Lastly relevant results were analyzed with Cytoscape 3.8.0. AutoDock vina software was used for molecular docking to analyze the binding energy of the active components and key targets of Baihe Dihuang decoction. PyMOL software were used to visualize the optimal docking results. ICR male mice were randomly divided into control group, model group, Rolipram group, low, medium and high dose group of Baihe Dihuang decoction. After 14 days of administration, the neurobehavioral scores of mice in each group were collected, and the expression of related proteins in brain tissue was detected, ELISA and Western blotting were used to detect the expression of the key protein cAMP, PKA, p-CREB and BDNF. At last, the adverse reaction of Baihe Dihuang decoction was observed by vomiting experiment. RESULTS A total of 13 active components and 39 key targets were collected from network pharmacology. The docking results showed that the first 10 core targets all performed well and their effects were closely related to PRKACA. Compared with the control group, the model group mice's recognition rate of new objects and the spontaneous alternation reaction rate were significantly reduced, the escape latency was significantly prolonged, and the target quadrant stay time, the number of crossing platforms were significantly reduced; cAMP, PKA, p-CREB and BDNF in the hippocampus of mice was significantly decreased. Baihe Dihuang decoction could reverse the behavior of AD mice and the expression of cAMP, PKA, p-CREB and BDNF. In the vomiting experiment, the anesthesia recovery time of the Rolipram group was significantly prolonged, while that of the Baihe Dihuang decoction group was not significantly affected. CONCLUSION The mechanism of Baihe Dihuang decoction in the treatment of AD may be related to its influence on cAMP-PKA and regulation of cAMP-PKA-CREB-BDNF signal pathway, and the adverse reactions are milder than those of clopramide.

Objective To investigate the characteristics of changes in interleukin(IL)-33 and regulatory T(Treg)cells in brucellosis,to verify the regulatory effect of IL-33 on Treg cells,so as to clarify the immune mechanism of IL-33 on Treg cells in brucellosis.Methods The peripheral blood of 39 patients with brucellosis treated in the People's Hospital of Xinjiang Uygur Autonomous Region from January to December 2021(the brucellosis group)and 42 healthy controls(the healthy control group)who underwent physical examination during the same period were collected.The serum IL-33 level was detected by AimPlex kit,and the proportion of Treg cells was detected by flow cytometry.Peripheral blood mononuclear cell(PBMC)was extracted and cultured in vitro to observe the proportion and mRNA expression levels of forkhead box protein P3(Foxp3)after stimulation and blocking of IL-33.Results Compared with the healthy control group,the level of IL-33 and the proportion of Treg cells in brucellosis group were significantly increased,with statistical significance(P＜0.05).In vitro tests showed that the Foxp3 proportion and mRNA expression level of PBMC in the two groups were significantly increased after IL-33 stimulation,and significantly decreased after IL-33 blocking,with statistical significance(P＜0.001).Conclusion IL-33 and Treg cells increased significantly in brucellosis patients,and IL-33 promoted the immune function of Treg cells.Blocking IL-33 is expected to be a potential target for immunotherapy of brucellosis.