Effects of ethane 1,2-dimethanesulfonate preconditioning on renal ische-mia/reperfusion injury in male rats / 中国病理生理杂志

AIM:To investigate the effects of ethane 1,2-dimethanesulfonate (EDS) preconditioning on renal ischemia/reperfusion (I/R) injury in male Sprague-Dawley (SD) rats.METHODS: Male SD rats (n=48) were ran-domly assigned to 6 groups:blank, sham, I/R, EDS+I/R, EDS+testosterone (TST) +I/R, and castration (Cast)+I/R.The renal pedicles were bilaterally occluded with a microvascular clamp for 45 min to establish renal I/R-induced in-jury model.Bilateral orchiectomy was conducted 2 weeks before surgery .EDS (75 mg/kg) was intraperitoneally injected 5 d before operation .Blood samples were collected 24 h after reperfusion from the vena orbitalis posterior plexus .Luteinizing hormone (LH), TST, serum creatinine (SCr), blood urea nitrogen (BUN), and kidney injury molecule-1 (KIM-1) were detected.The renal tissues were harvested to measure the level of TNF-αand the expression of Fas mRNA and caspase-3 protein.RESULTS:Serum TST levels in EDS +I/R group and Cast +I/R group were below the minimum detectable threshold.Compared with other groups , the rats in EDS+I/R group and Cast +I/R group had higher levels of SCr , BUN and KIM-1 (P0.05), but KIM-1 level in EDS+I/R group was lower than that in Cast +I/R group (P<0.05).After reper-fusion for 24 h, the levels of TST and LH in EDS +I/R group, Cast+I/R group and EDS+TST+I/R group were lower than those 1 h before operation (P<0.05).Compared with Cast+I/R and I/R group, the TNF-αlevel and expression of Fas mRNA and caspase-3 protein were significantly decreased in EDS +I/R group ( P<0.05 ) .CONCLUSION: EDS preconditioning substantially reduces the serum TST level , thus attenuating I/R-induced acute renal injury .TNF-α-induced Fas/FasL pathway may be involved in this process .

Clinical analysis of 30 cases of drug induced liver injury / 中国基层医药

Objective To investigate the clinical features and occurrence regularity of drug induced liver injury,so as to provide reference for clinical medication.Methods The choice of hospital 2012 -2014 reported adverse drug reaction monitoring center data,including DILI 30 cases,DILI related data to classify statistics,compre-hensive analysis.Results DILI was of many influence factors,the time of occurrence of large differences,partial onset occult,and related to a drug to immune enhancement agent most(23.33%),followed by anti -tumor drugs (20.00%)and lipid regulating agent(16.67%).Conclusion The medical staff should enhance the understanding of DILI medicine source disease,strengthen the monitoring of adverse drug reactions,as far as possible to reduce the occurrence of DILI,and make its harm to a minimum,reduce the patients pain and medical expenses of disease.

Generalized design of fitness equipment control interface / 中国组织工程研究

BACKGROUND:Health has been paid more and more attention in modern society, the health of those special groups, such as the elderly and the disabled, should also be brought to the forefront. As fitness equipment is more and more popular in contemporary, our focus should be transferred to the use of fitness equipment for these special groups. OBJECTIVE:To put forward the design strategy of fitness equipment control interface, for the special groups to use common fitness equipment comfortably and conveniently. METHODS:Based on the research of existing fitness equipment control interface, according to the usage habits and cognitive awareness of special groups on control interface, a new design strategy has been put forward using the general design method and ergonomics. RESULTS AND CONCLUSION:Based on the existing 45 kinds of exercise bikes, 84 kinds of running machines and 129 kinds of treadmil fitness equipments, we found that these equipments are designed for common users in the aspects of design and production process and control interface. Universal design is a design concept beyond the lack of capacity to meet the different needs of different populations, and designed products can reduce or even eliminate the difference in the ability of special populations and the general population in use. The new strategy makes design of fitness equipment control interface generalized and good man-machine interactive, which can meet the demand of the special groups and help them do fitness exercise to improve their health and body function.

Man-machine interface design of a rehabilitation training equipment / 中国组织工程研究

BACKGROUND:Rehabilitation training equipments play an important role in the rehabilitation treatment. Because of poor muscle strength and joint mobility in patients, we must guarantee the safety of rehabilitation training equipments. OBJECTIVE:To design a new suitable man-machine interface that ensures patients can use rehabilitation equipments and even parts of fitness equipments safely. METHODS:Through user experience research, we found the flaws of the existing rehabilitation equipment. Depending on the principles of ergonomics, we designed a new man-machine interface for upper limb exercise through survey and computer-aided design. RESULTS AND CONCLUSION:The new man-machine interface program achieves the rapid wear and discharge between patients and rehabilitation training equipment, and importantly, it can automatical y separate people from the equipment when the patient's body discomforts or equipment failure appears. What’s more, this man-machine interface can be promoted to other fitness equipments. As a result, rehabilitation training for patients wil be more convenient.

Digital three-dimensional model reconstruction of the cardiac cavity / 中国组织工程研究

BACKGROUND:Digital three-dimensional model which can reflect the fine structure of the chambers inside heart not only enhances the understanding of cardiac physiology, but also provides basic medical data for the study of cardiac electrophysiology simulation and endocardial electrophysiological mapping navigation. OBJECTIVE:To construct the digital three-dimensional model of cardiac cavity from sectional data and in conformity with the actual anatomical structure. METHODS:Image segmentation was accomplished in MATLAB environment. Firstly, registration of human cardiac cavity slice dataset was realized. Secondly, classifying each composition was achieved by clustering method according to color characteristics of the image. Then, both cardiac cavity and related connected region was distinguished by region growing method. At last, the processed image was reconstructed through dedicated medical processing software into three-dimensional model of the cardiac cavity. RESULTS AND CONCLUSION:The proposed method could reconstruct quite exquisite three-dimensional model of the cardiac cavity. In models, left and right atrial and ventricular structure was clear. Aorta and superior vena cava were visible. Three tricuspid and mitral valve were also observed. Results indicated that reconstructed model can reflect the anatomical characteristics of cardiac cavity accurately, and provide basic medical data for the study on electrophysiological simulation and endocardial electric mapping.

Simulation analysis and experimental study of positioning signals in thorax electric field catheter / 生物医学工程学杂志

In order to enhance the position accuracy of ablation catheter in heart electrophysiology operation, signals of respiration and heartbeat must be removed for subsequent data processing. Based on locating principle of electrical field with low frequency, synchronous detector with MC1496 has been developed in this study. In the present research, several methods are utilized to optimize the circuit performance, such as coupling and stopping direct current, low-pass filtering, as well as limiting ripple voltage etc. Through simulation results, it showed that the demodulation performance of the circuit was fine. Through simulation platform of thorax electric field and animal experiment, the circuit feasibility were further proved good for extracting signals of respiration and heartbeat.

Pharmacokinetics of a fusion protein for human acidic fibroblast growth factor and transcriptional activator protein in rat and its penetration across blood-brain barrier / 药学学报

This paper is to report the study of the pharmacokinetics of a fusion protein TAT-haFGF(14-154) for human acidic fibroblast growth factor and transcriptional activator protein in rat plasma, and the investigation of their penetration across blood-brain barrier in mice and rats, in order to provide a basis for clinical development and treatment of Alzheimer's disease. Enzyme-linked immunosorbent assay (ELISA) was used to determine concentration of TAT-haFGF(14-154) in rat plasma and in mouse brain homogenate; and immunohistochemistry was used to analyze the distribution in brain. The concentration-time curve fitted two-compartment open model which was linear kinetics elimination after a single intravenous injection of TAT-haFGF(14-154) in rat at the dose of 300 microg x kg(-1). The half life time was 0.049 +/- 0.03 h for distribution phase and 0.55 +/- 0.05 h for elimination phase, and the weight was 1/C2. The result showed that TAT-haFGF(14-154) could be detected in the brain by ELISA and immunohistochemistry, the elimination of TAT-haFGF(14-154) in rat was swift, and TAT-haFGF(14-154) could penetrate across the blood-brain barrier, distribute in pallium and hippocampus and locate in the nucleus.

Preparation and in vitro/in vivo pharmacological studies of bFGF/collagen composite sponge / 中国药科大学学报

Aim: To investigate a new type wound dressing,basic fibroblast growth factor(bFGF)/collagen com-posite sponge,and conduct its pharmacological studies in vitro and in vivo.Methods: bFGF/collagen composite sponge was prepared using fresh pig skin and bFGF.The sponge's physicochemical properties were studied.MTT assay was used to detect the proliferation effect of the sponge extract on 3T3 cells.Delayed allergy of the sponge was tested for the assurance of its biosafety.Results: Results showed that the physicochemical properties of bFGF/collagen composite sponge with high and low doses of bFGF have no significant difference from those of blank collagen sponge.SDS-page analysis indicated that the composite sponge has apparent strip in 18 kD.It was also found that bFGF/collagen composite sponge was responsible for significant effects on 3T3 cell proliferation in comparison to saline treatement(P <0.01,P <0.05).In the allergy study,during the periods of the induction and stimulation,no allergic reaction was found in bFGF/collagen composite sponge groups with high and low doses of bFGF,while severe reactions and inflammation occurred in positive group(2,4-dinitrochlorobenzene).Furthermore,pathological examination indicated the intact dermal structure and no sign of inflammation.Conclu-sion: The developed sponge has good physicochemical propertis and noticed cellular proliferation without dermal irritation.There is much potential to develop bFGF/collagen composite sponge into a new kind of wound dressing material for clinical use.

Effect of basic fibroblast growth factor plus Brucea Javanica oil emulsion on repairing back skin wound of rabbits / 中国组织工程研究

BACKGROUND: It has been proved that the application of basic fibroblast growth factor (bFGF) combined with Brucea Javanica oil emulsion can accelerate wound healing and inhibit scar formation.OBJECTIVE: To observe the effects of bFGF plus Brucea Javanica oil emulsion cream on accelerating the skin wound healing of rabbits.DESIGN: A randomized controlled observation.SETTING: Center of Biotechnological Research and Development, Jinan University; College of Pharmacy, Jinan University.MATERIALS:The experiment was carried out in the College of Pharmacy, Jinan University from June to September in 2004. Eight Beijing big-ear white rabbits (4 males and 4 females) of 2.0-2.5 kg were provided by the experimental animal center of Southern Medical University (certification number: SoKx-2002-010). bFGF sterile freeze dried powder agent,provided by Guangzhou Changsheng Gene Pharmaceutical Co.,Ltd (batch number: 20040219; specific activity was 6 000 U/bottle), was prepared to solution with water for injection before application. Brucea Javanica oil emulsion (manufactured by Zhejiang 999 Bang'erkang Pharmaceutical Co.,Ltd.) was provided by Professor Yao from staff Room of Pharmacy, Shenyang Pharmaceutical University.METHODS: The rabbits were anesthetized and disinfected, 5 round wounds with diameter of 1.8 cm and area of 2.54 cm2were induced from front to back by bilateral incision at 1.5 cm from middle spine of rabbit. The 5 wounds of each rabbit were randomly divided into bFGF-treated group (90 U/cm2), bFGF+Brucea Javanica oil emulsion group [the wound was smeared with Brucea Javanica oil emulsion (30 mg/cm2) 30 minutes after bFGF (90 U/cm2)], Brucea Javanica oil emulsion treated group [the wound was smeared with Brucea Javanica oil emulsion (30 mg/cm2)], blank emulsion group (30 mg/cm2) and blank control group (the wound was smeared with saline). The medication was give immediately after injury, and changed once a day for 16 days. At 4, 8, 12 and 16 days after injury, the wound areas were recorded with the method of hyaline membrane tracing (the wound was covered with clean saran wrap, the size of wound was traced,and then sheared to be weighed, and converted to calculate the area), and the volume of wounded cavity was measured by infusing water. At 8 and 16 days, the wound tissue was removed, stained after routine tissue sections, and the conditions of growth of granulation tissue and reepithelization on the wound surface were observed.MAIN OUTCOME MEASURE: The wound area, volume of wounded cavity, and the conditions of growth of granulation tissue and reepithelization on the wound surface were obviously at different time points after injury in each group.RESULTS:All the 8 rabbits were involved in the analysis of results. ① Wound areas at different time points in each group: The wound areas in the bFGF+Brucea Javanica oil emulsion group at 4, 8 and 12 days after medication were smaller than those in the blank control group at corresponding time points [(2.05±0.35), (1.59±0.25), (0.55±0.25) cm2;(2.53±0.30), (2.41±0.19), (1.09±0.34) cm2, P＜0.05-0.01]. The wound areas in the bFGF group at 8 and 12 days after medication were (1.71±0.31) and (0.51±0.10) cm2, which were significantly smaller than those in the blank control group at corresponding time points (P＜0.05-0.01). ② Volume of the wounded cavity in each group: The wound volume in the bFGF group at 4 days after injury was markedly smaller than that in the blank control group at corresponding time point [(0.49±0.12), (0.59±0.1) mL, P＜0.05]. The wound volumes in the bFGF+Brucea Javanica oil emulsion group at 4 and 8days after injury were significantly smaller than those in the blank control group at corresponding time points [(0.47±0.12), (0.30±0.08) mL; (0.59±0.1), (0.41±0.07) mL, P＜0.05, 0.01]. ③ Growth of granulation tissue and reepithelization on the wound surface in each group: At 8 days after injury, the inflammatory reaction was milder and fibroblasts proliferated significantly in the bFGF+Brucea Javanica oil emulsion group, and the numbers of capillary plumules and fibroblasts were significantly more than those in the blank control group. The conditions in the blank control group and blank emulsion group were generally the same that there were severe inflammatory reactions, obvious increase of granulation tissue, fewer new capillaries, and unobvious proliferation of epidermic cells. At 16 days after injury, the contraction and reepithelization on the wound surface were obvious, and the new epithelia went towards the wound center rapidly in the bFGF+Brucea Javanica oil emulsion group.CONCLUSION : The application of Brucea Javanica oil emulsion plus bFGF can obviously accelerate the repair of incised wound on the back of rabbits.

A Convenient and Effective Approach for Gene Synthesis of Human Platelet Factor-4 / 中国药科大学学报

AIM:To synthesize the human platelet factor-4(hPF4) gene with a convenient and effective approach, and high express the hPF4 gene in E. coli BL21 (DE3). METHODS: According to the primary structure of hPF4, the nucleotide sequence was synthesized using touch-down PCR method. The resultant gene fragment containing EcoR Ⅰ and Xho Ⅰ overhangs at 5' and 3' ends was cloned into the expression vector pGEX-4T-1 to construct the recombinant plasmid pGEX-4T-1-hPF4,which was then transformed into the E. coli strain BL21 (DE3). RESULTS: hPF4 gene was successfully synthesized by touchdown PCR method. A fusion protein composed of glutathione S-transferase (GST) and the recombinant hPF4 was expressed in BL21(DE3) by IPTG induction. The expression level of the fusion protein in E. coli was about 30% of the total cellular protein. CONCLUSION: Touch-down PCR may provide a convenient and effective approach to obtain other target genes. The expressed fusion protein forms the inclusion bodies, providing sufficient material for further purification and biological activities process.

Expression and Purification of a hbFGF Lacking Nuclear Localization Signal / 中国药科大学学报

AIM:To study the mechanism of the unique export of one of human basic fibroblast growth factor (hbFGF) forms lacking the N-terminal nuclear localization signal (NLS),we high expressed and purified this hbFGF form in E.coli strain BL21(DE3).METHODS:The cDNA fragment of the hbFGF amplified by polymerase chain reaction (PCR) was cloned into the expression vector pET3c and expressed in BL21(DE3) by IPTG induction.The expressed hbFGF was purified by ionic exchange and heparin affinity chromatography from the supernatant of bacteria lysate.The mitogenic activity was measured by MTT.RESULTS:The expression level of hbFGF in E.coli was about 20% of the total cellular protein.The appreciable mitogenic activity of the purified hbFGF was comparable to that of hbFGF standard.CONCLUSION:The BL21(DE3)/ pET3c expression system could be used to high express hbFGF lacking NLS.The purified recombinant hbFGF was prepared and sufficient for further study.

Expression and Purification of Basic Fibroblast Growth Factor Mutant with Reduced Mitogenic Activity / 中国生物工程杂志

In order to decrease the potential side-effects of human basic fibroblast growth factor (hbFGF) caused by its broadspectrum mitogenic activity, a single residue of hbFGF, the residue serine 108, was replaced with neutral alanine residue to construct a mutant of hbFGF (mhbFGF) with reduced mitogenic activity. The mutant was overexpressed in Escherichia coli BL21(DE3) by IPTG induction. The expression level of mhbFGF was about 30% of the total cellular protein. The expressed mhbFGF was purified by ionic exchange and heparin affinity chromatography from the supernatant of bacteria lysate. Measured by MTT method, the effect of mhbFGF on Balb/c 3T3 cell proliferation was much lower than that of wild-type hbFGF. The purified recombinant mhbFGF was prepared and sufficient for the following pharmacological and safety studies.

Protective effects of nmhaFGF on NRK52E cell apoptosis induced by H_2O_2 / 中国病理生理杂志

AIM:The present study was designed to establish H2O2-induced NRK52E cell apoptotic model and to investigate the effects and the mechanism of nmhaFGF on the NRK52E cell apoptosis induced by H2O2. METHODS: In vitro experiment, a apoptotic model of normal rat kidney epithelium (NRK52E) was made by MTT method, Hoechst33342 dyeing and flow cytosorting (FCR). NRK52E cells were cultured with different concentrations of nmhaFGF and haFGF for 24 h before H2O2 was added. The apoptotic rate was detected with FCR method. RESULTS: H2O2 at concentration of 0.4 mmol/L, the optimal condition to establish the apoptotic model, was used to treat NRK52E cells for 18h. Different doses of nmhaFGF (0.01, 0.03, 0.10, 0.30, 1.00 mg/L) reduced the apoptotic rates with the dose rising. However, the decreasing tends of apoptotic rates with dose increasing for haFGF was not so obvious. CONCLUSION: nmhaFGF protects the NRK52E cells against apoptosis. The mechanism might be connected with its non-mitogenic property.

The mitogenic activity decline of a haFGF mutant and its mechanism / 中国病理生理杂志

AIM: To observe the effects of the human acidic fibroblast growth factor mutant (mhaFGF), lacking 27 amino acids at N-terminal, on the proliferation and signal transduction of the hepatocarcinoma cells. METHODS: The hepatocarcinoma cells were treated with human acidic fibroblast growth factor (haFGF) and mhaFGF, respectively. The expression levels of the signal proteins, Grb2 and Erk1/2, in the hepatocarcinoma cells were detected by semi-quantitative Western-blotting after treated for 15 min. The mitogenic activity of both haFGF and mhaFGF was detected by MTT method and the cell cycle was analysed by flow cytometer (FCM) after treated for 48 h. RESULTS: The mitogenic activity and the ratio of G 1 and S phase cells in mhaFGF-treated cells were markedly lower than that of the haFGF, and close to that of the control group. The expression level of both Grb2 and Erk1/2 in the mhaFGF-treated cells were lower than those in the haFGF- treated cells. CONCLUSION: The decrease in the mitogenic activity of mhaFGF is probably associated to its down-regulating the expression of the signal molecular, MAPK-ERK1/2 and Grb2.