Objective To observe vancomycin and vancomycin in elderly patients with renal toxicity.Methods 105 cases because of infection from March 2013 to October 2014 were collected and randomly divided into two groups, one had 52 patients and were given vancomycin for anti-infection treatment, another group had 53 patients and were given norvancomycin for anti-infective treatment.Changes of serum urea nitrogen and creatinine levels and adverse reactions were observed and compared between two groups.Results Creatinine levels of patients with vancomycin group after 10 days and 7 days after withdrawal were (97.86 ±8.27)μmoI/L, (82.03 ±5.72)μmoI/L, and the norvancomycin group were (98.67 ±8.34)μmoI/L, (83.47 ± 5.91)μmoI/L, the difference were not significant.Urea nitrogen levels of patients with vancomycin group after 10 days and 7 days after withdrawal were (6.71 ±1.15)mmoI/L,(6.09 ±1.09)mmoI/L, respectively, and the norvancomycin group were(6.75 ±1.17)mmoI/L,(6.15 ±1.12)mmoI/L, the difference were not statistically significant.The total effective rate of vancomycin group was 78.85%, and norvancomycin group was 75.47%, the difference was not statistically significant.Adverse reactions of vancomycin group during treatment was 13.46%, and norvancomycin group was 13.21%, the difference was not statistically significant.Conclusion Vancomycin and norvancomycinboth have anti-infective effect on renal function in patients with certain adverse effects, urea nitrogen, creatinine levels in two groups were elevated during treatment, but decreased after withdrawing medicine.

Objective Infrared spectroscopy was used to study on both raw material and different degree water processing varieties extract of Radix Aconiti kusnezoffii,to observe the changes of main toxic components in processing,and thus to improve the quality of Radix Aconiti kusnezoffii processed products.Methods Fu Liye transform infrared spectroscopy was adopted to study the infrared spectrum characteristic of raw materials and different degree water processing varieties of.Radix Aconiti kusnezoffii.Results ①Aconitine and hypaconitine infrared spectrum showed that 1717 cm-1,1727 cm-1 and 1711 cm-1 is the C=O stretching vibration peak.It is a diester alkaloids characteristic peaks; ② although absorption peak of all vesicular samples had a certain change,it still existed diester alkaloids absorption peak,indicating the incomplete hydrolysis; ③ boiled aconite processing methods demonstrated diester alkaloids absorption peak shift in the water sample.Conclusion Diester alkaloids in Radix Aconiti kusnezoffii shows a positive relation with its time soaked in water.

Objective To apply iTRAQ technology to observe changes in protein expression group in the process of inducing C2C12 cells differentiation towards osteoblast by BMP-2.Methods The myoblast C2C12 cells were seeded in BMP-2 induced differentiation system for differentiation induction.In the 7th day,differentiation protein was extracted and labeled with iTRAQ reagent.Then,mass spectrometric detection,data analysis of differentially expressed proteins,and analysis of biological information were carried out.Results 23 significantly differentially expressed protein spots were screened by BMP-2-induced myoblast C2C12 differentiated cell protein expression profile analysis,where the protein was labeled with iTRAQ reagent.8 protein points were up-regulated,and 15 protein points were down-regulated.Trend classification found that the above differential protein had differential expression in each period of C2C12 cell osteogenic differentiation (1-7 days).Part of up-regulated protein in the early differentiation period showed high expression level;part of up-regulated protein in the late differentiation period showed high expression level;similarly,part of down-regulated protein in the early differentiation period presented low expression level;part of down-regulated protein in the late differentiation period showed low expression level.Preliminary identification showed SERCA3,Cytochrome bS,S100A4,ATPase inhibitor and ATPIF1 presented dynamic changes,which suggests that these proteins may be related to inducing osteogenic differentiation mechanism.Conclusion The results of differential protein expression trend show the necessity of full monitoring of C2C 12 cells osteogenic differentiation and indicate that iTRAQ technology is an effective method of studying protein changes of cellular molecule.Five proteins including SERCA3,Cytochrome b5,S100A4,ATPase inhibitor and ATPIF1 can be used as candidate targets for osteogenic differentiation mechanism research.

Objective To investigate the mechanism of oxidative damage caused by lipopolysaccharide (LPS)induced sepsis in rat brain.Methods The rats were randomly divided into control group and model group (low LPS group and high LPS group).Twenty-four hours after the modeling,the rats were sacrificed before their brain tissue was taken out and prepared for the test.The changes in malondialdehyde (MDA),superoxide dismutase (SOD),glutathione peroxidase (GSH-Px),total antioxidant capacity (T-AOC),hydrogen peroxide (H2 O2 )and succinate dehydrogenase (SDH)were detected.The expression level of JNK and Nrf2 protein in brain tissue was detected by qRT-PCR and Western blotting. Results Compared with the control group,the MDA,SOD,GSH-px,T-AOC,H2O2 and SDH level increased significantly in the model group,and the difference in expressions of JNK and Nrf2 was statistically significant (P <0.05). Conclusion The LPS induced septic oxidative brain damage model in rats is successfully established,and the process may be regulated through the Nrf2 and JNK signal pathways.

Objective To investigate the protective effect of rosiglitazone on the rats with high altitude pulmonary edema.Methods Thirty-six SD rats were randomly (random number) divided into 6 groups (n =6 each):control group (Control),hypobaric hypoxia model group (HH),rosiglitazone groups (RSG) which were administered with 3 different doses [RSG-L:5 mg/ (kg · d),RSG-M:10 mg/ (kg·d),RSG-H:20 mg/ (kg· d)],dexamethasone group [Dex,4 mg/ (kg· d)].Rats were injected intraperitoneally with different doses of rosiglitazone (RSG),dexamethasone (Dex) or vehicle (Control and HH) for 3 days before placed in simulated altitude of 6 000 m hypobaric hypoxia animal chamber where the temperature and pressure were constant.After 72 h in the chamber,each rat was anesthetized.The water content of lung was determined with wet/dry weight ratio.Bronchoalveolar lavage fluid was measured by bradford method.The contents of GSH was measured by micro-ezymed labeled method.The contents of MDA was measured by TBA method.The enzymatic activities of SOD was measured by WST-1 method.The changes of the TNF-α,IL-6 and IL-10 in serum were determined by ELISA.Light microscope was used to observe the pathological changes of lung tissue.Results Compared with Control group,the wet/dry weight ratio of lung (5.08 ± 0.24) and total protein content of BALF (351.06 ± 44.55) μg/mL increased significantly (P ＜ 0.01) in HH group.There were red blood cells in the alveolar and interstitium,pink fluid exudation in the alveolar,the alveolar septum enhancement,and a large number of inflammatory cell infiltration;the SOD activity (10.65 ± 0.94) U/mgprot and the content of GSH (1.63 ±0.20) μmol/gprot in lung tissue were significantly decreased (P ＜ 0.01),the contents of MDA (2.1 5 ± 0.18) nmol/mgprot increased significantly (P ＜ 0.01),TNF-o (56.92 ± 2.87) pg/mL and IL-6 (217.80 ±48.01) pg/mL levels in serum were significantly increased (P ＜0.01),and IL-10 (76.85 ± 16.72) pg/mL level decreased (P ＜ 0.05).Compared with the HH group,the wet/dry ratio of lung and total protein content of BALF in different doses of rosiglitazone group significantly decreased (P ＜ 0.01),the pathological changes of the lung tissue was significantly improved,SOD activity and the content of GSH in lung tissue was significantly increased (P ＜ 0.01),the content of MDA decreased (P ＜ 0.01),The levels of TNF-α and IL-6 in serum were significantly decreased (P ＜ 0.01),while the IL-10 level was significantly increased (P ＜ 0.01).Conclusion Rosiglitazone could protect the high altitude pulmonary edema by alleviating the oxidative stress and inflammatory response.

OBJECTIVETo investigate the influence of drug properties on the encapsulation effiency (EE) and drug release of transfersomes for a proper transfersome preparation.

METHODTo prepare the transfersomes of colchicines (CLC), vincristine sulfate (VCR) and mitoxantrone hydrochloride (DHAD) with the same materials and methods, and then measure their EE. To find out the relationship between drug properties like solubility, molecular weight and charges, and EE. To performe the drug release experiments of various types of transfersomes in vitro, and compare their differences.

RESULTVCR and DHAD are lipophilic or hydrophilic, owing positive charges and large molecular weight, as a result, their EE are high, while CLC is amphipathic, neutral, and of small molecular weight, its EE is very low. As DHAD can insert into the membrane of transfersome, the drug release of DHAD-T in vitro is much slower than that of VCR-T.

CONCLUSIONTo prepare transfersomes with high EE, drugs that are lipophilic or hydrophilic, high molecular weight and opposite charges to the membrane should be chosen. Interaction between drugs and membrane will influnce the rate of drug release.

Antineoplastic Agents ; administration & dosage ; chemistry ; Antineoplastic Agents, Phytogenic ; administration & dosage ; chemistry ; Colchicine ; administration & dosage ; chemistry ; Deoxycholic Acid ; Drug Carriers ; Gout Suppressants ; administration & dosage ; chemistry ; Mitoxantrone ; administration & dosage ; chemistry ; Particle Size ; Phosphatidylcholines ; Solubility ; Technology, Pharmaceutical ; methods ; Vincristine ; administration & dosage ; chemistry

BACKGROUND: Vascular endothelial growth factor (VEGF) is a very effective way to make tissue engineer bone vascularization.However, because of expensive and short half-life, VEGF cannot maintain effective concentration in blood after injection. To resolve the problem effectively, gene transfection technique is used in this experiment to transfer human VEGF into seed cells-mesenchymal stem cells (MSCs) of tissue engineer bone and to make it secrete VEGF which could vascularize bone.OBJECTIVE: To explore the possibility of human vascular endothelial growth factor 165 (VEGF165) to transfect rabbit MSCs, and establish the experimental foundation of angiogenesis tissue engineering organization and the treatment of ischemic disorders.DESIGN: Observation control trail.SETTING: First Hospital of Jilin University and Institute of Frontier Medical Sciences of Jilin University.MATERIALS: The experiment was conducted in the Key Laboratory (BSL-2) of Frontier Medical Sciences of Jilin University between June 2003 and August 2004. Health New Zealand white rabbits, 4.0-5.0 months old, weighing 2.5-3.5 kg, half male and half female, were provided by Animal Center of Jilin University. The rabbits were handled under asepsis and anesthetized condition,corresponding to the animal ethical standard. Medicine and reagents: Ham F12 culture media (Gibco, U.S), MTT (Sigma, U.S)PLXSNKDRp-VEGF165 and pcDNA 3.0 vectors were prepared in the present laboratory. ELISA detection kit (Jingmei company,Shenzhen), DH5 α, restriction endonucleases Barn H I, Xhol Ⅰ, Hind Ⅲ, EcoR Ⅰ and standard DNA molecule (Promega,U.S) were also used in this study.METHODS: Rabbits' MSCs were separated and cultivated. The pcDNA 3.0-hVEGF165 expression vector was constructed and identified, pcDNA3.0-VEGF165 eukaryotic expression vector was constructed, the vector was used directly to transfect MSCs. The cultural supernatant then was collected and the soluble protein of human VEGF gene expression was analyzed with ELISA method.The proliferation capability of human umbilical vein endothelial cells (HUVEC) stimulated by the supernatant was measured with MTT methods, untreated MSCs and pcDNA3.0 transfected MSCs were used as control groups.MAIN OUTCOME MEASURES: ① Result of restriction enzyme digestion and DNA sequencing of the recombinant plasmid pcDNA3.0-VEGF165;② the secretion of human VEGF165 proteins of the transfected MSCs analyzed by ABC-ELISA; ③ MTT method was used to detect the effects of MSCs culture supematant transfected with VEGF165 on HUVEC cells proliferation ability.RESULTS: ①Result of restriction enzyme digestion and DNA sequencing of the recombinant plasmid: The constructed plasmid was digested with Hind Ⅲ and XHol Ⅰ, and then two pieces fragments were isolated with agarose gel electrophoresis, which was accordance with expected results. And sequencing results showed that PeDNA3.0-VEGF165 eukaryotic expression vector was successfully constructed. ② ABC-ELISA method: Compared with the control group, concentration of human VEGF protein in the supernatant of the cultured cells increased significantly after the MSCs were transfected with pcDNA3.0-VEGF165 for 24, 48, 72 hours (P＜0.05).③ MTT method was used to detect the effects of MSCs culture supernatant transfected with VEGF165 on HUVEC cells proliferation ability. The results showed MSCs supematant transfected with VEGF165 (2%, 4%,8%, 16%, and 32%) had statistical significance in promoting HUVEC cells proliferation rate compared with the normal control (P＜0.05).CONCLUSION: Human VEGF gene can be successfully transfected into MSCs and expressed effectively.

Objective To establish a mouse model of prostate cancer expressing human PSCA for the development of new anti-tumor drugs or vaccines. Methods The total RNA of DU145 cells,a human prostate cancer cell line,was isolated by using TRIzol reagent according to the (RT-PCR),the first-strand cDNA was synthesized using the SuperScript First-Strand synthesis system. The human PSCA gene was amplified with the primers and cloned into the plasmid pcDNA3.1 to generate pcDNA-PSCA. DNA sequencing was used to confirm the constructs. The mouse prostate tumor cell line RM-1 cells,syngeneic to C57BL/6,were transfected with pcDNA-PSCA plasmids followed by selection using G418. RT-PCR analysis was performed to examine the validity of the constructs. Expression of PSCA on the cell surface was determined by staining with anti-PSCA antibody,and the anti-PSCA antibody was detected using an FITC-conjugated goat anti-rabbit IgG antibody,and analyzed by flow cytometry. 4-6-week-old male C57BL/6 mice purchased from the Laboratory Animals Center were inoculated with different amounts of RM-PSCA cells to search for suitable cell population which can form tumor in mouse,and the mice were monitored twice a week. The growth and the survival time of mice were measured,respectively. The tumor volume was measured by vernier caliper according to the formula:V=0.5a×b~2,where a and b are the long and short diameters of the tumor,respectively. Results The plasmid pcDNA-PSCA was successfully constructed and the PSCA was successfully expressed in RM-PSCA 7~# and RM-PSCA 28~# cells by RT-PCR and confirmed by flow cytometry. 1×10~5 RM-PSCA cells were sufficient to get tumor growth in 100% of inoculated mice. The tumor grew quickly and the volume of the tumor reached 12000 mm~3 within 34 days. All the mice died within 40 days and their mean survival time was 37 days. Conclusion A PSCA-expressing tumor model in mice has been successfully established. It can be used to evaluate the activities of drugs or vaccines.

Objective To prepare the anti-TLR4 C-terminal domain monoclonal antibody and investigate its effect in treatment of sepsis.Methods TLR4 C-terminal polypeptide (amino acid sequence:368-579,named as TLR4-C) was obtained through prokaryotic expression and Sephacryl S-100 gel purification,and then was used to immunize female Balb/c mice (6-8 weeks old).After cell fusion,antibody screening and purification,monoclonal antibody specific for the C terminal of TLR4 was obtained.Specificity of monoclonal antibody was detected by Western blot and cell immunofluorescence.In vitro antibody activity test,NR8383 was cultured for 1 h with adding antibody (100 μg/ml) and then 12 h after adding lipopolysaccharide (LPS) (10 ng/ml),and level of tumor growth factor (TNF)-α in the culture medium was tested by ELISA.In vivo septic animal experiment,40 SD rats were assigned to control antibody group (n =20) and anti-TLR4 monoclonal antibody group (n =20) according to the random number table.Each group was rejected 50 mg/kg corresponding antibodies via caudal vein for 1 h,and then LPS (10 mg/kg) via intraperitoneal injection for 4 h.Blood samples from caudal vein of ten rats in each group were collect to test the serum level of TNF-α.The rest rats in each group were used to measure the animal survival rate within 72 h.Results Three highly specific anti-TLR4 monoclonal antibodies were obtained and could combined with TLR4-C and TLR4 holoprotein.In vitro cell activity study indicated only one monoclonal antibody could obviously inhibit the release of TNF-α.In vivo animal experiment showed serum TNF-α level in anti-TLR4-C antibody group was (1.54 ± 0.18) ng/ml,significantly lower than (0.51 ± 0.10) ng/ml in antibody control group (P ＜ 0.01).Animal survival rate in anti-TLR4-C antibody group was 70％,higher than 30％ in antibody control group (P ＜ 0.05).Conclusion Anti-TLR4-C monoclonal antibodies have great capacity to neutralize TLR4 and good protective effect on LPS-induced sepsis.

Objective To study the effects of dynamic,support-inducing exercise on the support,balance and gait ability of patients with moderate-to-severe hemiplegia after stroke.Methods Fourteen stroke patients were randomly assigned to an experimental or a control group (7 cases to each).The patients in the experimental group received both dynamic,support-inducing exercise and routine exercises,while the patients in the control group received routine exercises only.Before training and after 40 and 60 days of training,their functional capacity was evaluated with the Chinese stroke scale (CSS) for neurological deficits,Berg's balance scale (BBS) and using functional ambulation categories (FACs).Results Before training there was no inter-group difference in average CSS or BBS scores or in FACs.For the experimental group there were significant intra-group differences compared with 0th day in all three items at both time points.At days 40 and 60 there were also significant intra-group differences in BBS scores and FACs in the control group,and CSS scores improved significantly only in the experimental group.At day 40 there were significant inter-group differences in average CSS,BBS and FAC results.However,by day 60 a significant difference persisted only in average CSS scores.Conclusions Dynamic,support-inducing exercise can improve support,balance and gait in patients with moderate-to-severe hemiplegia after stroke.