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 evaluate the accuracy of air column width difference (ACWD) between inflation and deflation of the cuff of the endotracheal tube (ETT) in predicting post-extubation stridor (PES).Methods A total of 102 intubated patients of both sexes and all ages,who were mechanically ventilated for ≥24 h in the intensive care unit,of American Society of Anesthesiologists physical status Ⅲ or Ⅳ,were enrolled in the study.After the patients were in a stable condition and recovered consciousness and myodynamia,they were weaned from the ventilator,and ultrasound examination of the larynx was performed to determine ACWD between ETT cuff inflation and deflation.PES was assessed using blinding nethod after extubation,and the patients were divided into 2 groups depending on whether or not the patients developed PES:PES group (group P) and non-PES group (group N).Results There were 94 patients in group N and 8 patients in group P.Compared with group N,ACWD was significantly decreased in group P (P＜ 0.05).The cut-off value of ACWD determined by the receiver operating characteristic curve was 1.65 mm,the sensitivity and specificity of ACWD were 0.830 and 0.750,respectively,and the area under the receiver operating characteristic curve was 0.801.Conclusion For the intubated patients who are mechanically ventilated for ≥24 h in an intensive care unit,ACWD between ETT cuff inflation and deflation＜ 1.65 mm can effectively predict PES.

Objective To analyze clinical manifestation and investigate therapy effect of re-operation of tricuspid regurgitation (TR) after mitral valve replacement with rheumatic heart disease.Methods Seventeen cases with rheumatic heart disease recurred TR after mitral valve replacement surgery,underwent tricuspid valve surgery again in the Third People's Hospital of Nanchang from January 2000 to December 2011.Of 17 cases,10cases underwent tricuspid valve annuloplasty including 1 case for pure De Vega plasty,9 cases for the valve leaflets forming + artificial valve ring forming.Another 7 cases underwent tricuspid valve replacement surgery including 4 cases for biological valve replacement and 3 cases for mechanical valve.Retrospective analyzed the clinical manifestations,treatment process and condition of prognosis.Results One case was with early postoperative deaths (5.88％,1717),and died of postoperative left ventricular failure.Three cases were postoperative low cardiac output syndrome,2 cases were renal insufficiency,and 2 cases were respiratory insufficiency,all those cases were successfully cured.Sixteen cases were followed up from 3 months to 9 years and 2 cases were lost.Of 14,2 cases were NYHA class Ⅰ,8 cases for grade Ⅱ,4 cases for grade Ⅲ.Conclusion After mitral valve replacement in patients with rheumatic heart disease,TR in patients with reoperation is a suitable choice.Reasonable surgical indications,timing of surgery and good perioperative management are the keys to improve the success rate of surgery.

Objective To evaluate the feasibility and effectiveness of the application of IQQA (Intelligent/interactive Qualitative and Quantitative Analysis) three-dimensional reconstruction technique in precise laparoscopic or robot-assisted laparoscopic partial nephrectomy (LPN or RAPN) for renal hilar tumors.Methods The study retrospectively reviewed 11patients with hilar tunors from February 2016 to February 2017.Of the 11 patients,4 were women and 7 were men,with an average age of 51 years (range 38 to 70 years).The average tumor size was 3.1 cm (range 1.7 to 4.3 cm).For tumor stage,9 patients were in T1a stage and 2 patients were in T1b stage.Tbe average R.E.N.A.L score was 8.7 (range 7 to 10).The mean preoperative GFR was 40.6 ml/min (range 32 to 45 ml/min).IQQA three-dimensional reconstruction technique was applied for the purpose of precise navigation and resection of the tumors.Multivariate analysis was used to identify predictors of warm ischemia time,estimated blood loss,major perioperative complications,and postoperative renal function.Results All 11 laparoscopic or robot-assisted laparoscopic hilar partial nephrectomies were successfully completed without conversion to a hand-assisted or an open approach.Under the navigation of IQQA,all tumors were found precisely at the first time during surgeries.The final pathologic examination revealed that all the 11 patients were clear cell renal cell carcinomas.The mean operative time was 142 minutes (range 90 to 230 minutes),with a mean warm ischemia time of 24 minutes (range 17 to 33 minutes).The estinated blood loss was 146 ml (range 50 to 400 ml).No intraoperative complications occurred.Two patients suffered from postoperative complications.One patient with gross hematuria was recovered by consistent bladder irrigation.The other patient with postoperative hemorrhage needed transfusion.All patients had negative margins on the final pathologic examination.At a mean follow-up period of 3 months,the mean GRR is 22.5 ml/min (range 13 to 34 ml/min) without any disease recurrence.Conclusions With peculiar features,such as accurate location,complete resection and fewer perioperative complications,the application of IQQA three-dimensional reconstruction technique in precise partial nephrectomy represents a safe and effective procedure for hilar tumors.

The paper presented the " Professors Evaluation System" initiated by the hospital since July 2009, which was designed for quantitative assessment of senior medical professionals of non-management category in terms of their performance in medical, teaching, research and management work. Seven periods of appraisal have witnessed a significant elevation of the rate of excellence among such professionals, proving that such a system can motivate and guide these professionals to continuously improve their competence in medical, teaching, research and management work.

Particulate wear debris within the bone-prosthesis microenvironment generated by normal wear and corrosion of orthopaedic implants is considered to be one of the main factors responsible for chronic aseptic inflammation and development of osteolysis in the long-term instability and failure of total joint arthroplasty. While the decrease in bone volume caused by wear debris-induced osteolysis could have been compensated by enough new bone matrix secreted by osteoblasts. Actually, the normal osteoblastic population depend on the regular differentiation and proliferation of their progenitor cells--bone marrow mesenchymal stem cells (MSCs). This study aims to investigate the potential mechanism for the rat MSCs cytotoxicity upon exposure to Titanium (Ti) particles. Rat mesenchymal stem cells (rMSCs) isolated from 3-month-old male Sprague-Dawley rats by Percoll intensity gradient method were cultured in DMEM medium (low glucose) supplemented with 10% fetal bovine serum, 100 U/ml penicillin, and 100 micrograms/ml streptomycin in a humidified incubator with 5% CO2 at 37 degrees C. In order to gain the homogenous cell population, rMSCs were passaged to 3-4th subpassage which were used in all the experiment groups. Then rMSCs were seeded in the 6 well culture plates and exposed to three different circle diameters (mean size, TD1: 0.9 micron, TD2: 2.7 microns, TD3: 6.9 microns) with three different concentrations (0.1 wt%, 0.05 wt%, 0.01 wt%, W/V) at different durations (8 h, 16 h, 24 h,), respectively. Unexposed rMSCs were used as control. In the given periods of Ti loading, fluid shear stress (FSS) was applied to each group cells. The expression of F-actin and DNA of the rMSCs at the indicated time were determined with laser confocal scanning microscopy and image analysis software. The results showed that there was up-regulation expression of F-actin in the rMSCs without Ti particles loading but in the presence of FSS. Ti particles loading can suppress the expression of F-action and DNA of rMSCs, but this down-regulation response varied with the three circle diameter, concentrations and durations of Ti particles. Among three kinds of diametrically different Ti particles, submicron Ti particles (0.9 micron) had the greatest suppressive response on rMSCs, together with some apoptosis bodies. Under the same diameter condition, the inhibition induced by Ti particles loading was in a manner dependent on the particles concentration and exposure duration. The reductive effects produced from 0.1 wt% Ti was the greatest and earliest among the responses from Ti particles at three different concentrations; and the lower the concentration, the weaker the repressive influence. Furthermore, with the elongation of exposure to Ti particles, the expression of F-actin and DNA decreased gradually, the lowest level was at 32 h. These findings demonstrated that Ti particles loading can attenuate rMSCs' viability in a manner dependent on the circle diameter, particles concentration, treatment period, suggesting that a reduction in the number of viable MSCs together with a compromise of the their differentiation into functional osteoblast may exacerbate aseptic loosening of total joint implant. Further investigation into particles-mediated suppression of MCSs viability may reveal novel mechanism of implant loosening and aid in development and application of osteolytic drug therapy and the optimization of design and selection of future orthopaedic biomaterials, thereby improving long-term compatibility and stability for arthroplasty patients.
Actins ; metabolism ; Animals ; Cells, Cultured ; DNA ; metabolism ; Male ; Mesenchymal Stromal Cells ; cytology ; drug effects ; metabolism ; Particle Size ; Rats ; Rats, Sprague-Dawley ; Stress, Mechanical ; Time Factors ; Titanium ; administration & dosage ; toxicity

MicroRNAs have been identified as a new class of regulatory molecules that affect many biological functions by interferring the target gene expressions. Latest studies demonstrate that microRNAs can influence many pivotal bio-processes and deeply involve in the metabolism of glucose, lipid and amino acid and biological oxidation. For glucose metabolism, microRNAs are related to insulin secretion, insulin sensitivity, glucose uptake, glycolysis, oxidation and mitochondrial function. For lipid matebolism, microRNAs can regulate the target genes related to lipid biosynthesis, catabolism and transportation. MicroRNAs can influence glutamine catabolism.
Animals ; Glucose ; metabolism ; Glutamine ; metabolism ; Humans ; Insulin ; metabolism ; Insulin Secretion ; Lipid Metabolism ; physiology ; Metabolism ; physiology ; MicroRNAs ; physiology

Objective To investigate the protective effect of Carbachol on hyperoxia-induced acute lung injury (HALI) in mice and its related mechanisms. Methods Thirty-two healthy male ICR mice were randomly divided into four groups:control group, hyperoxia exposure three days group (HO3d group), hyperoxia exposure three days + Carbachol group (HO3d+Carba group), and Carbachol group (Carba group), eight mice in each group. The pathological changes of lung tissue in each group were observed under light microscope after the models were completed in each group.The expression of TLR4 and NF-κB protein in lung tissues were detected by Western blot, and the expression of HMGB-1 and TNF-α mRNA in lung tissues by RT-PCR. LSD-t test was used for sample pairwise comparison, and one-way ANOVA for intergroup comparison. P<0.05 was considered statistically significant. Results There was no statistical difference between the control group and the Carba group (P> 0.05), and no obvious abnormal changes in lung tissue structure. The expression of TLR4, NF-κB protein and HMGB-1 and TNF-α mRNA in the HO3d group were significantly higher than those in the control group (P<0.01), and there were obvious bleeding on the surface of the lung tissue and severe pathological damage. The expression of TLR4,NF-κB protein and HMGB-1 and TNF-α mRNA in the HO3d+Carba group were significantly lower than those in the HO3d group(P<0.01), while lung tissue damage degree was also lower than that in the HO3d group. Conclusions Hyperoxia can increase the expression of TLR4 and NF-κB in lung tissues, and cause inflammatory injury in lung tissue. Carbachol can reduce the release of HMGB-1 and TNF-α inflammatory factors in hyperoxia-induced acute lung injury, and its mechanism is related to the inhibition of TLR4/NF-κB signal pathway, which has a protective effect on HALI.

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, Mammalian ; Eukaryotic Cells ; metabolism ; Gene Expression ; Genetic Vectors ; Humans ; Kidney ; cytology ; metabolism ; Peroxidases ; biosynthesis ; genetics ; Peroxiredoxin III ; Peroxiredoxins ; Plasmids ; genetics ; Transfection

ObjectiveTo study the clinical efficacy of Shire Biqing pill in the treatment of rheumatoid arthritis （damp-heat obstruction syndrome） and its effect on the expression of serum osteoprotegerin （OPG）， nuclear factor-κB receptor activating factor ligand （RANKL）， and tumor necrosis factor-α （TNF-α）， and to explore its mechanism from the perspective of bone destruction. MethodPatients with rheumatoid arthritis （damp-heat obstruction syndrome） were randomly divided into two groups， with 36 patients in each group. The control group was treated with methotrexate tablets and celecoxib capsule， while the treatment group was treated with Shire Biqing pill based on the control group. The treatment period was 3 months. The pain visual analogue scale （VAS） score， joint tenderness number， joint swelling number， disease activity score （DAS28-ESR）， traditional Chinese medicine （TCM） symptom quantitative score， and related adverse reactions were recorded before and after treatment， and the peripheral serum OPG， RANKL， TNF-α， erythrocyte sedimentation rate （ESR）， and Creactive protein （CRP） were detected. ResultAfter treatment， the total effective rate was 88.57% （31/35） in the treatment group and 79.41% （27/34） in the control group. The total effective rate of the treatment group was higher than that of the control group （Z=-2.089， P<0.05）. The pain VAS score， joint tenderness number， joint swelling number， and DAS28-ESR of the two groups were significantly lower than those before treatment （P<0.05）， and the pain VAS score， joint tenderness number， joint swelling number， and DAS28-ESR of the treatment group were significantly better than those of the control group after treatment （P<0.05）. Compared with that before treatment， the TCM symptom quantitative score in the two groups decreased significantly （P<0.05）， and the decrease was more obvious in the treatment group than in the control group （P<0.05）. Compared with those before treatment， the levels of RANKL， TNF-α， ESR， and CRP in the two groups decreased and the level of OPG increased （P<0.05）， and the changes in the treatment group were more obvious that in the control group （P<0.05）. There were no serious adverse events or serious adverse reactions during this clinical trial. ConclusionShire Biqing pill can effectively improve the clinical symptoms of rheumatoid arthritis （damp-heat obstruction syndrome） with good safety. Shire Biqing pill effectively regulate the OPG/RANKL/RANK system and reduce the pro-inflammatory factor TNF-α， which may be its mechanism in the intervention in rheumatoid arthritis bone destruction.