Objective To construct Gadd45a expression plasmid and induce its expression in human T cells.Methods Gadd45a was amplified by reverse transcription PCR from human embryonic stem cells,and cloned into the pcDNA3.1 vector.The recombinant plasmid or blank plasmid was transfected into Jurkat cells or normal human CD4+T cells using electroporation,and the expression of Gadd45a was detected by quantitative RT-PCR and Western blot.Results Human Gadd45a expression plasmid was constructed successfully.Gadd45a was overexpressed both in Jurkat cells and normal human CD4+T cells after these cells were transfected with pcDNA3.1-Gadd45a.Conclusion The construction of Gadd45a expression plasmid and induction of Gadd45a overexpression in human T cells lay the foundation for further research on the role of Gadd45a in the epigenetic mechanism.

Objective To explore the possible association of oxidative stress reaction with the mRNA expression of podocin in rats with adriarnycin(ADR)-induced nephrotic syndrome.Methods mRNA expression of podocin in renal cortex were investigated by in situ hybridization staining and semi-quantitative RT-PCR in ADR-induced nephrotic rats,the level of malondialdehyde(MDA), superoxide dismutase(SOD)and total antioxidative capacity(T-AOC) in renal cortex were measured, then the relationship between them was evaluated.Results In ADR rats, MDA increased at d7 and reached a significant higher level at d14 and d21;SOD decreased at d14 and persisted to d21; T-AOC decreased at d21.The podocin mRNA mostly expressed in cytoplasm of glomerular cells. Both the number and the intensity of positive cells increased notably as time progressed in ADR rats.The level of podocin mRNA expression showed no obvious changes at d7, while prominently increased at d14 and pesisted to d21 in ADR rats.There was significant correlativity between the mRNA expression of podocin and the level of MDA,SOD,T-AOC.Conclusion The mRNA expression of podocin is correlated with oxidative stress reaction in rats with ADR-induced nephrotic syndrome.

Objective To explore the effects of UVB on the expression of Gadd45a gene and DNA methylation levels in Jurkat cells. Methods Jurkat cells were irradiated with UVB of 1.0 J/cm2 and 1.5 J/cm2 respectively, and collected at 6, 12, 24 and 48 hours after the irradiation. Real-time RT-PCR was used to detect the mRNA expression of Gadd45a gene and methylation-sensitive genes CD11a and CD70. Global methylation level was also measured by MethylAmp global DNA methylation quantification kit. Results After irradiation with UVB at 1.0 J/cm2, the mRNA level of Gadd45a increased but global methylation level decreased at 6, 12, 24 and 48 hours, and significant changes were observed at 6 and 12 hours for the level of both Gadd45a mRNA expression and global methylation (P ＜ 0.01 or 0.05). Elevated mRNA expressions of CD11 a and CD70 were also noted in Jurkat cells after irradiation with UVB of 1.0 J/cm2, and significant elevation was observed at 12 hours (both P ＜ 0.05 ). After irradiation with UVB of 1.5 J/cm2, there was a statistical increase in the mRNA expressions of Gadd45a, CD11 a, CD70, together with a statistical decrease in global methylation level in Jurkat cells, at 6, 12, 24 and 48 hours (P ＜ 0.01 or 0.05). The mRNA expression of Gadd45a negatively correlated with the global level of DNA methylation in Jurkat cells (r = -0.395, P ＜ 0.05). Conclusion UVB irradiation can upregulate the expression of Gadd45a, but downregulate the global DNA methylation level in Jurkat cells.

Objectives To explore the effects and mechanism of acupuncture treatment in emergent of acute hypertension by studying the acupuncture treatment on acute hypertension rats. Method The acute hypertension rat model was made by injecting adrenalin into the abdomen. After probing specific points, NO, ET-1, angiotensin and rennin levels were tested. Results Acupunctured for 30 minutes, the experimental group's diastolic pressure dropped and NO level rose, and these changes were statistically significant regarding those of the medicine control group. After the acupuncture treatment, the rennin level of the medicine control group and the experimental group were statistically different with the control group, but no significant difference was found between the medicine control group and the experimental group. Conclusion Probing Quechi, Taichong, Sanyinjiao and Neiguan may lead to the improved performance of the vessel endoderm, the rise of NO level and the decrease of angiotensin content, and consequently reduce the blood pressure of acute hypertension rats.

Objective To investigate the effect of dihydromyricetin on proliferation and apoptosis of breast cancer MCF-7 cells.Methods From March 2014 to February 2015,breast cancer MCF-7 cells were treated with 99% pure DMY as an inhibitor.MTT assay,flow cytometry and immunocytochemistry were used to analyze the proliferation,apoptosis and protein expression of breast cancer cell MCF-7.Results When the DMY concentration was higher than 20 μg/mL,the inhibitory effect appeared,but not good.When 40 and 80 μg/mL DMY were used,the proliferation of MCF-6 cells were significantly inhibited,and have different degrees of sensitivity to it.When DMY was 80 μg/mL,the IC50 was 226.9 μg/mL.The inhibition rate and IC50 were compared with 0 μg/mL DMY,there was significant difference(P<0.05).Indicating that the relatively high concentration of DMY could significantly improve the patient′s condition of breast cancer;MCF-6 cells treated with 40 and 80 μg/mL DMY could induce G2/M phase arrest in DMY with concentration greater than 20 μg/mL(P<0.01),and showed significant cell apoptosis death phenomenon.At the same time,G0/G1 phase was also blocked and S phase cells decreased significantly,the difference was significant(P<0.05).The positive rate of PCNA in breast cancer MCF-6 cells significantly decreased(P<0.01) when the DMY concentration was higher than 50 μg/mL,the positive rate was> 50%,especially in DMY with 80 μg/mL,the positive rate was 10.00%.Compared with 0 μg/mL DMY,the difference was significant(P<0.05).Conclusion The use of dihydromyricetin in breast cancer patients can effectively inhibit the rapid increase of cancer cells,accelerate apoptosis,slow down the patient′s condition,the effect is outstanding.

Objective To observe the preventive effect of alprostadil on contrast-induced nephropathy (CIN) in patients undergoing coronary intervention.Methods Two hundred and fifteen patients undergoing coronary intervention were divided into treatment group (112 cases) and control group (103 cases) by random digits table method.Patients in treatment group were treated routinely with addition of alprostadil (10 μ g intravenous injection every 12 h once for 7-10 days,given before surgery).Patients in control group were given routine therapy only.Complications of the two groups and CIN occurrence were observed.Results The occurrence rate of CIN in treatment group was lower than that in control group[7.1％ (8/112) vs.18.4％ (19/103),P ＜ 0.05].The occurrence rate of CIN in patients with renal insufficiency was higher than that in patients with normal renal function [51.6％ (16/31) vs.6.0％ (11/184),P ＜ 0.01].The occurrence rate of CIN in patients with diabetes was higher than that in patients without diabetes [22.1％(15/68) vs.8.2％ (12/147),P ＜ 0.01].Conclusions Alprostadil in coronary intervention before treatment can obviously reduce the incidence of CIN.CIN is more likely to occur in patients with renal insufficiency or with diabetes.

Objective To explore the risk factors for infection following percutaneous nephrolithotomy(PCNL), and provide the basis for prevention of postoperative infection.Methods 96 patients who were performed PCNL in a hospital between August 2010 and August 2013 were chosen,clinical data of patients were retrospectively analyzed, the occurrence of postoperative infection,risk factors and isolated pathogens were analyzed.Results 35 patients (36.46%[35/96])developed infection following PCNL.Univariate analysis revealed that risk factors for postopera-tive infection were the size and shape of stone,duration of operation,and intraoperative perfusion (all P <0.05). 96 patients before operation and 18 patients after operation were performed bacterial culture for urine,a total of 39 strains were isolated,the main isolated pathogens was Escherichia coli (n=12),followed by Pseudomonas aerugi-nosa (n=8),Streptococcus viridans (n =5 ),Klebsiella pneumoniae (n =4)and Enterobacter cloacae (n =3 ). Conclusion Infection rate following PCNL is high,duration of operation and intraoperative perfusion should be re-duced,antimicrobial agents should be used rationally.

AIM To study toxicity of Safflor Yellew (SY) for vascular endothelic cell (VEC) and whether SY has antagonistic effect on platelet activating factor (PAF). METHODS Use light microscopical observation and crystal violet colorimetric method to study morphological and bioactivity change of HVEC and EVC 304 after incubated with PAF, SY, SY+PAF, Ginkolide,and Ginkolide+PAF. RESULTS Cultured with PAF 30 minutes, HVEC had histopathological changes: from fusiform, polygond squamous cell to ellipsoid, circular cell , not binding tightly, but separated by wide gap. As cultured with SY+PAF, cell's morphological changes just as same as PAF's group. And its bioactivity had tendency of decline. Either HVEC was incubated with SY (0.571 g?L -1 ) for 30 minutes or EVC 304 cultured with SY(1 143 g?L -1 ) for 4 hours, their morphology and bioactivity had changes. CONCLUSION SY do not inhibit PAF's injury effect. And high concentration SY has toxicity to cultured endothelic cell.

Objective To study the correlation between chemotherapy drug sensitivity and the expression of P-glycoprotein protein(P-gp),lung resistant-related protein(LRP)in different esophageal carcinoma patients,which will offer experimental evidence for clinical individualized chemotherapy of esophageal carcinoma patients.Methods Selected fresh esophageal carcinoma's tissue,Common used drugs and chemoscheme for esophageal carcinoma were used for chemosensitivity testing of the primary cultured tumor cells.At the same time,immunohistochemistry method was used to detect the expression of P-gp and LRP of esophageal carcinoma cells.Results The chemotherapy drug sensitivity of single drug was different(P＜0.05).The combination of Tax and DDP had the best effect.DDP and NVB combination had a better effect than DDP and 5-FU combination.The sensitivity of DDP had no significant correlations with expression intensity of P-gp(P＞0.05),and it had significant correlation with expression intensity of LRP(P＜0.05).There were significant correlations between sensitivity of both NVB group and Tax group and expression intensity of P-gp and LRP(P＜0.05).There was no significant correlations between sensitivity of 5-FU and expression intensity of P-gp and LRP(P＞0.05).Conclusion The esophageal carcinoma chemosensitivity in vitro can indicate the sensitive drug of different patients,and guide individualized chemotherapy.The expression of P-gp and LRP has relationship with several chemotherapy drugs,which can provide base for individualized chemotherapy in clinic.

Air pollution and global warming are increasingly deteriorating. Large amounts of polyamides derived from fossil fuel sources are consumed around the world. Cadaverine is an important building monomer block of bio-based polyamides, thus biotechnological processes for these polymers possess enormous ecological and economical potential. Currently, the engineered strains for biological production of cadaverine are Corynebacterium glutamicum and Escherichia coli. We review here the latest research progress of biosynthesis of cadaverine including metabolism of cadaverine in microorganisms, key enzymes and transport proteins in cadaverine synthesis pathway, optimum pathways and cadaverine yields.
Biosynthetic Pathways ; Biotechnology ; Cadaverine ; biosynthesis ; Corynebacterium glutamicum ; metabolism ; Escherichia coli ; metabolism