To construct the eukaryotic expression plasmid of human PRX3 and measure its expression in the HEK-293FT cells, the full-length coding region of human PRX3 was cloned by PCR and inserted into the eukaryotic expression vector pcDNA4-Xpress (A). HEK-293FT cells were transiently transfected with the recombinant plasmid. Western blot and immuofluorescence were used to detect the expression of the fusion protein. In the experiment, restriction analysis identified the construction of the recombinant plasmid and the inserted sequence was identical with that published on GenBank. Western blot and immunofluorescence confirmed the expression of the recombinant protein in transfected HEK-293FT cells. It was concluded that the eukaryotic expression plasmid of human PRX3 was constructed successfully and the recombinant could be expressed efficiently in HEK-293FT cells, which provides a sound basis for the further study on human PRX3.
Cell Line, Transformed ; Cloning, Molecular ; Embryo ; Eukaryotic Cells/*metabolism ; Gene Expression ; Genetic Vectors ; Kidney/cytology ; Kidney/*metabolism ; Peroxidases/*biosynthesis ; Peroxidases/genetics ; Plasmids/*genetics ; Transfection

Objective To investigate the role and intracellular signal transduction mechanism in the injury of renal tubular cells induced by postasphyxial-serum in neonate.Methods Human renal proximal tubular cell(HK-2 cell) was used as target cell. The experiment was designed as:control group, asphyxia group ,and pyrrolidine dithiocarbamate (PDTC)blocking group. The attacking concentration of serum was 20%, and the apoptosis rate of HK-2 cells was detected by flow cytometer.Results Compared with controls[(13.3?1.70)%],after being stimulated with postasphyxial-serum, the apoptosis rate of HK-2 cells of asphyxia group [(46.73?3.68)%] and PDTC blocking group [(31.19?2.79)%]were significantly increased(P

Objective To investigate the role of postasphyxial-serum of neonate in inducing injury of human renal proximal tubular cells(HK-2 cells).Methods HK-2 cells were used as target cell.The neonatal different concentration postasphyxial-serum of 1,3,7 days after asphyxia were used as attacking means.The experimental groups were divided into 15 groups:the 2.5%,5.0%,10.0%,(20.0%) attacking concertration groups of 1,3,7 day after asphyxia and control group of each concertration.The culture medium and concertration of the control group and the experimental groups were the same.The changes of morphology were observed under inverted microscope,the cell viability was measured by 3-(4,5-dimethyl-2-thiazoly1)-2,5-diphenyl-2H-tetrazolium bromide(MTT) method and the leakage rate of lactate dehydrogenase(LDH) was determined by biochemical methods.Results Compared with control group,the changes in morphology of HK-2 were most serious and obvious,the cell viability were obviously decreased(all P

OBJECTIVETo compare clinical therapeutic effect of surrounded needling therapy and medication on acute gouty arthritis.

METHODSSixty cases of acute gouty arthritis were randomly divided into a surrounded needling therapy group and a western medicine group, 30 cases in each group. The surrounded needling therapy group was treated with shallow needling on local affected area as main and 4-5 distant acupoints as adjuvant, once each day; the western medicine group was treated with oral administration of Indomethacin and Allopurinol, thrice each day. They were treated for 15 days. The clinical therapeutic effects, the changes of serum uric acid content and the adverse reaction were observed in the two groups.

RESULTSThe total effective rate was 93.3% in the surrounded needling therapy group and 80.0% in the western medicine group, with a significant difference between the two groups (P < 0.01); the serum uric acid contents before and after treatment were (516.85 +/- 48.63) micromol/L and (293.77 +/- 68.45) micromol/L in the surrounded needling therapy group, and (509.66 +/- 51.11) micromol/L and (333.66 +/- 89.22) mciromol/L in the western medicine group, respectively, with significant differences before and after treatment in the two groups (both P < 0.05), and with a significant difference in the serum uric acid content after treatment between the two groups (P < 0.01). The surrounded needling therapy group had no adverse reaction, and the adverse reaction rate of the western medicine group was 46.7%, with a significant difference between the two groups (P < 0.01).

CONCLUSIONSurrounded needling therapy is superior to the western medicine in the therapeutic effect on acute gouty arthritis, and it is a safe and effective method for acute gouty arthritis.

Acupuncture Points ; Acupuncture Therapy ; Acute Disease ; therapy ; Adult ; Aged ; Allopurinol ; therapeutic use ; Arthritis, Gouty ; blood ; drug therapy ; therapy ; Humans ; Indomethacin ; therapeutic use ; Male ; Middle Aged ; Treatment Outcome ; Uric Acid ; blood

OBJECTIVETo observe the promoting consciousness effect of electroacupuncture combined with routine western medicine therapy on the patient with coma caused by craniocerebral trauma.

METHODSThirty-two cases were randomly divided into an acupuncture-medication group treated with electroacupuncture at Neiguan (PC 6) and Quze (PC 3) and routine western medicine, and a control group treated with routine western medicine, 16 cases in each group. Glasgow (GCS) scores were assessed after treatment for 7 sessions and 30 sessions respectively and the promoting consciousness rate was observed.

RESULTSAfter treatment of 7 sessions, GCS score was 6.88 +/- 1.63 in the acupuncture-medication group and 5.25-1.65 in the control group with a significant difference between the two groups (P < 0.05); after treatment of 7 sessions, the promoting consciousness rate was 25.0% in the acupuncture-medication group and 0 in the western medicine group, and after treatment for 30 sessions, the promoting conscious ness rate was 81. 3% in the acupuncture-medication group and 43.8% in the western medicine group with a signifi cant difference between the two groups (P < 0.05).

CONCLUSIONElectroacupuncture at Neiguan (PC 6) and Quze (PC 3) combined with western medicine has a good promoting consciousness effect in the patient with coma caused by craniocerebral trauma, which is better than that of simple western medicine.

Adolescent ; Adult ; Aged ; Coma ; drug therapy ; etiology ; therapy ; Combined Modality Therapy ; Craniocerebral Trauma ; complications ; Electroacupuncture ; Female ; Humans ; Male ; Middle Aged ; Young Adult

Biosimilars are biological medicinal products that are highly similar to an already licensed reference product in terms of quality, safety, and efficacy. BAT1706 is being developed by Bio-Thera Solutions, Ltd. as a proposed biosimilar candidate to bevacizumab reference product (Avastin^®). Bevacizumab acts by specifically binding to vascular endothelial growth factor A (VEGF-A), and preventing the interaction of VEGF-A with its receptors on the surface of endothelial cells, then blocking the downstream signaling pathway mediated by ligand-receptor, and inhibiting endothelial angiogenesis, thus inhibiting tumor growth. Comprehensive analytical characterization studies incorporating orthogonal analytical techniques were performed to compare the in vitro functional activities of BAT1706 and Avastin^®. BAT1706 and Avastin^® showed highly similar binding activity to multiple VEGF-A isoforms and equivalent VEGF-A neutralizing activity, as well as inhibitory activity of VEGF receptor (VEGFR)-2 tyrosine kinase autophosphorylation. Both products exhibited similar binding of the Fcγ receptors and a lack of Fc-related effector functions such as antibody-dependent cell-mediated cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC). Overall, the results demonstrate that BAT1706 and Avastin^® are highly similar in terms of in vitro functional activities.

Adult ; Female ; Humans ; Male ; Medial Collateral Ligament, Knee ; injuries ; surgery ; Middle Aged ; Reconstructive Surgical Procedures ; methods ; Rupture ; Suture Anchors

Placentation,which is critical for maternal-fetal exchange of nutrients and gases,is a complicated process comprising stepwise vasculogenesis and angiogenesis.Hypoxia caused by impairedtrophoblast invasion may cause various angiogenic abnormalities in human placenta.The Notchl signaling pathway plays an important role in the regulation of angiogenesis.The angiogenesis of human umbilical vein endothelial cells (HUVECs) under normal/hypoxic conditions and the mRNA/protein level of Notchl/Dell4/Jaggedl were investigated in this study.The effects of DAPT/JAG-1 on the migration of HUVECs were also assessed by cell wound healing assay,so as to discover the possible role of notchl signaling pathway in the angiogenesis of human placenta.The results showed that angiogenic ability of HUVECs was seriously reduced under hypoxic conditions.The mRNA and protein levels of Notchl/Dell4/Jaggedl were decreased in the hypoxic group compared to the control one.In addition,the migration capability of HUVECs was significantly obstructed when treated with DAPT and under hopoxic condition,but promoted when treated with JAG-1.The above results demonstrate that hypoxia downregulates the angiogenesis in human placenta via Notch 1 signaling pathway.

The study was aimed to investigate the synergistically effect of interferon-α (IFN-α) and homoharringtonine (HHT) on the proliferation, apoptosis, cell cycle of K562 cells and the expression of β-catenin. The proliferation, apoptosis, cell cycle and β-catenin mRNA expression of K562 cells treated with IFN-α and/or HHT were assayed with MTT, flow cytometry or RT-PCR respectively. The results showed that HHT alone, but not IFN-α alone, displayed a proliferation inhibition, apoptosis induction, G(0)/G(1) phase block and down-regulation of β-catenin expression in K562 cells with concentration- and time-dependent manners. The expression level of β-catenin mRNA after being treated with HHT was 0.5576 ± 0.0373, which were lower than that in control group (0.9369 ± 0.0142). The down-regulation of β-catenin expression in group of IFN-α combined with HHT was higher significantly than that in HHT group (0.3737 ± 0.0529 vs 0.5576 ± 0.0373, P < 0.05). Otherwise, HHT combined with IFN-α did not demonstrate obvious toxicologic effect on bone marrow mononuclear cells. It is concluded that IFN-α combined with HHT can enhance the cytotoxic effect of HHT on K562 cells, which may be associated with down-regulation of β-catenin expression.
Cell Proliferation ; drug effects ; Harringtonines ; pharmacology ; Humans ; Interferon-alpha ; pharmacology ; K562 Cells ; beta Catenin ; genetics ; metabolism

AIMTo investigate the effect and mechanism of ascorbic acid on podocyte, last barrier of glomerular filtration, in diabetic rats.

METHODSDiabetic rats induced by streptozotocin injection intraperitoneally were treated by ascorbic acid for 5 weeks. The levels of blood glucose (BG), HbA1c, urinary albumin excretion rate (UAER) and superoxide diamutase (SOD), catalase (CAT) and malondialdehyde (MDA) in renal cortex were measured. The podocyte ultrastructure was observed while the expression of desmin protein, a marker of podocyte injury, was examined.

RESULTSCompared with control group, BG and HbA1c were increased markedly in diabetic group. The activities of SOD and CAT were decreased and the concentrations of MDA were increased significantly in diabetic renal cortex. There were the increased proteinic expression of desmin, foot process effacement in podocytes and UAER markedly in diabetic rats. Compared with diabetic rats, foot process effacement and the changes of UAER were ameliorated markedly while the activities of SOD were increased, the levels of MDA and proteinic expression of desmin were decreased markedly although BG, HbA1c and the activities of CAT were no significant difference in the diabetic rats by ascorbic acid treatment.

CONCLUSIONThe findings suggest that there are marked injury in podocyte, last barrier of glomerular filtration, in diabetic rats and administration of ascorbic acid can protect podocyte by increasing antioxidative capacity and ameliorating the renal oxidative stress.

Animals ; Ascorbic Acid ; pharmacology ; Catalase ; metabolism ; Desmin ; metabolism ; Diabetes Mellitus, Experimental ; metabolism ; pathology ; physiopathology ; Diabetic Nephropathies ; metabolism ; pathology ; physiopathology ; Male ; Oxidative Stress ; drug effects ; Podocytes ; drug effects ; metabolism ; ultrastructure ; Random Allocation ; Rats ; Rats, Wistar ; Superoxide Dismutase ; metabolism