To determine the number average molecular mass of chitosan-oligosaccharide by u-sing the acetylacetone's method based on the color-producing reaction with amino terminal . Via the tests on the repetitiveness, accuracy and the stability, supposed this method was reliable. At the same time, the contrastive results of HPLC and the traditional K3Fe(CN)6 method indicate that the acetylacetone method was more suitable for the determination of chitosan-oligosaccharide molecular mass.

Central composite design (CCD) is one of the most commonly used design methods in response surface optimization and has been widely applied in the field of pharmaceutics to optimize preparations. On the 20th anniversary of the introduction of CCD into China, the paper reviews its application in domestic pharmaceutical researches. Based on the brief introduction of basic principle and operation steps of CCD, the mistakes emerging in the application of CCD are summarized, including conceptual confusion with Box-Behnken design and face-centered CCD as well as wrong designs. Besides, the issues concerning the selection of factors and responses are discussed. The article is helpful for researchers to comprehensively understand the CCD and facilitates the rational application of this method.

Objective To investigate the morphological and functional alterations of peritoneum in uremic rats undergoing peritoneal dialysis(PD). Methods Thirty-five male Wistar rats were randomly divided into control group(sham operated group,n=6),uremia group(5/6 nephrectomy,n=6) and uremia with PD group(n=18).Uremia with PD group was subdivided into three subgroups according to different dialysis period(10 d,4 weeks and 8 weeks,n=6).Omenta were obtained for morphological examination,and peritoneal equilibration tests(PET) were performed to assess the transport function of peritoneal membrane. Results The number of blood vessels per high-power field in the uremia group,uremia with PD group and uremia with PD subgroups(5?3,10?5,17?5 and 19?4) were significantly increased compared with the control group(1?1),and that was much bigger in the uremia with PD group than the uremia group(P

BACKGROUNDIn tumors the process of apoptosis occurs over an interval of time after chemotherapy. It is important to determine the best time for detecting apoptosis by in vivo imaging. In this study, we evaluated the dynamics and feasibility of imaging non-small cell lung cancer (NSCLC) apoptosis induced by paclitaxel treatment using a (99)Tc(m)-labeled Annexin V recombinant with ten consecutive histidines (His10-Annexin V) in a mouse model.

METHODS(99)Tc(m)-His10-Annexin V was prepared by one step direct labeling; radio-chemical purity (RCP) and radio-stability was tested. The binding of (99)Tc(m)-His10-Annexin V to apoptotic cells was validated in vitro using camptothecin-induced Jurkat cells. In vivo bio-distribution was determined in mice by dissection. The human H460 NSCLC tumor cell line (H460) tumor-bearing mice were treated with intravenous paclitaxel 24, 48 and 72 hours later. (99)Tc(m)-His10-Annexin V was injected intravenously, and planar images were acquired at 2, 4 and 6 hours post-injection on a dual-head gamma camera fitted with a pinhole collimator. Tumor-to-normal tissue ratios (T/NT) were calculated by ROI analysis and they reflected specific binding of (99)Tc(m)-His10-Annexin V. Mice were sacrificed after imaging. Caspase-3, as the apoptosis detector, was determined by flow cytometry, and DNA fragmentation was analyzed by the terminal deoxynucleotidytransferase mediated dUTP nick-end labeling (TUNEL) assay. Nonspecific accumulation of protein was estimated using bovine serum albumin (BSA). The imaging data were correlated with TUNEL-positive nuclei and caspase-3 activity.

RESULTS(99)Tc(m)-His10-Annexin V had a RCP > 98% and high stability 2 hours after radio-labeling, and it could bind to apoptotic cells with high affinity. Bio-distribution of (99)Tc(m)-His10-Annexin V showed predominant uptake in kidney, relatively low uptake in myocardium, liver and gastrointestinal tract, and rapid clearance from blood and kidney was observed. The T/NT was significantly increased after paclitaxel treatment, whereas it was low in untreated tumors (T/NT = 1.43 ± 0.18). The %ID/g activity in Group 2 (24 hours), Group 3 (48 hours) and Group 4 (72 hours) after treatment was 2.55 ± 0.73, 3.35 ± 1.10, and 3.4 ± 0.96, respectively. Whereas in the non-treated group, Group 1, %ID/g was 1.10 ± 0.18. The radiotracer uptake was positively correlated to the apoptotic index (r = 0.852, P < 0.01), as well as caspase-3 activity (r = 0.816, P < 0.01).

CONCLUSIONThis study addresses the dynamics and feasibility of imaging non-small cell lung tumor apoptosis using (99)Tc(m)- His10-Annexin V.

Animals ; Annexin A5 ; Antineoplastic Agents, Phytogenic ; therapeutic use ; Apoptosis ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; pathology ; Cell Line, Tumor ; Disease Models, Animal ; Histidine ; Humans ; Lung Neoplasms ; drug therapy ; pathology ; Mice ; Organotechnetium Compounds ; Paclitaxel ; therapeutic use ; Radiopharmaceuticals

OBJECTIVETo observe the effect of blood activating and cooling, stasis removing herbs on the occurrence rate of Henoch-Schonlein purpura nephritis (HSPN).

METHODSThe 141 HSP children patients with bleeding of the blood stasis syndrome and of the blood heat syndrome (having normal results of urine routines) were assigned to the blood activating and stasis removing group and the blood cooling and arresting group. They were treated with blood activating and stasis removing herbs and blood cooling and arresting herbs respectively for eight weeks. The occurrence rate and time of the renal injury, changes of transforming growth factor (TGF), D-dimer (D-D), immunoglobulin (Ig), urine micro-protein, and urease before and after treatment were observed.

RESULTSThe occurrence of the renal injury in the blood activating and stasis removing group was 36.2%, obviously lower than that in the blood cooling and arresting group (69.4%). The occurrence time of the renal injury was 32.2 +/- 10.6 days, obviously later than that in the blood cooling and arresting group (20.0 +/- 9.0 days), showing statistical difference (P<0.05). The levels of TGF, D-D, IgA, microglobulin (MG), immunoglobulin G (IgG), albuminuria (ALB) of children patients in the blood activating and stasis removing group were lower after treatment than before treatment, showing significant difference (P<0.05). Significant difference was also shown when compared with those of the blood cooling and arresting group (P<0.05).

CONCLUSIONThe application of activating blood and removing stasis method could lower the probability of the renal injury in Henoch-Schonlein purpura (HSP). It played a role in preventing the occurrence of HSPN.

Child ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Incidence ; Kidney Diseases ; epidemiology ; Male ; Phytotherapy ; Purpura, Schoenlein-Henoch ; drug therapy ; epidemiology

Objective:To explore the protective effect of agmatine on mice with multiple organ failure (MODS) induced by yeast polysaccharide(ZYM) on the expression of inflammatory factors.Methods:ZYM induced inflammation model was established by intraperitoneal injection of ZYM in mice.All mice were divided into blank group,ZYM group and ZYM+AGM group.The mice feeding, white cell count,heart rate and so on were observed before and after the modeling to determine whether the model was successful.The liver function of mice,renal function,myocardial enzymes and other biochemical indicators were detected after the success of the model;and through the qPCR and ELISA method for detection of blood tumor necrosis factor alpha (TNF-alpha),interleukin 1 beta (IL-1 beta),interleukin 6 (IL-6),IL-10 gene and protein secretion level.Results: After the injection of ZYM,mice looked disorganized, activity and reduce consumption;the functional serological indexes of various organs of the mice were detected,which showed that the function of the viscera was serious.Compared with the blank control group,the serum parameters of ZYM group and ZYM+AGM group were significantly higher,and the inflammatory factors TNF-α,IL-1,IL-6 and IL-10 were significantly increased (P<0.05).Compared with ZYM group,ZYM+AGM serum markers of organ function decreased,inflammatory factor TNF-α,IL-1β,IL-6 decreased significantly (P<0.05),while there was no significant difference in IL-10 (P>0.05); and the mouse spirit,eating and activity had no significant change.Conclusion:Intraperitoneal injection of 500 mg/kg ZYM can successfully construct a model of MODS,AGM by reducing the release of inflammatory factors,play a protective role in the function of various organs of MODS mice.

BACKGROUND: Greatly importance has been attached to ceramic-on-ceramic bearing surface due to its excel ent wear resistance. But the risks of squeaking and ceramic fracture also go with it. Up til now, the choice between ceramic-on-ceramic and ceramic-on-polyethylene bearing surfaces in primary total hip arthroplasty remains controversial. OBJECTIVE: To compare the clinical outcomes and safety between ceramic-on-ceramic versus ceramic-on-polyethylene bearing surfaces in total hip arthroplasty based on meta analysis. METHODS: We electronical y searched databases including PubMed/Medline, Embase, Web of Science, Cochrane Col aboration database, Chinese Biomedical Literature Database (CBMdisc) and China National Knowledge Internet for randomized control ed trials on the comparison between ceramic-on-ceramic versus ceramic-on-polyethylene bearing surfaces in total hip arthroplasty from inception to January 2015. References of the included studies were also retrieved. Investigators severely selected the studies, extracted data and assessed the quality according to the inclusion and exclusion criteria. Then, meta-analysis was performed using RevMan 5.2 software. RESULTS AND CONCLUSION: Nine randomized control ed trials were included, involving 1 231 hips with ceramic-on-ceramic prosthesis and 932 hips with ceramic-on-polyethylene prosthesis. Meta analysis showed that both bearing surfaces achieved satisfied function recovery. But ceramic-on-ceramic had significantly increased risks of squeaking and ceramic fracture, meanwhile ceramic-on-polyethylene showed significantly higher wear rate. There were no significant differences in intra- or post-operative dislocation, osteolysis and other complications and prosthesis failure with any reason between two bearing surfaces. These results suggest that during the short- to mid-term fol ow-up period, no sufficient evidence can tel that ceramic-on-ceramic was obviously super than ceramic-on-polyethylene. Long-term fol ow-up is required for further evaluation.

Three kinds of Bacillus thuringiensis serotype-subsp. Leesis(H33) strain YBT-833, subsp. Aizawai(H7) strain YBT-1416 and subsp. Kurstaki(H3ab) strain YBT-1535, which were isolated by our lab, are chosen as original strain to clone vegetative insecticidal protein gene. Southern hybridization showed that vip genes are all localized at roughly 4-5 kb size-fractionated XbaI fragments of total DNA from YBT-833, YBT-1416 and YBT-1535. Three subgenomic libraries containing the vip gene fragment, were constructed with pUC19 as vector. Then, three vegetative insecticidal protein gene vip83, vip14 and vip15 are obtained from the libraries through the methods of colony-blot-in-situ screening and enzyme-cut detection. Comparision of DNA sequence made out that only vip83 gene exist five different base pairs with known vip genes. Because the sequences of vip14 and vip15 are the same, two of the three genes, vip83 and vip14, were subcloned to shuttle vehicle pHT315 to get recombinant plasmids pBMB8901 and pBMB8902 in turn. The plasmids were separately transformed into vip Bt. receptors BMB171 and 4Q7 to obtain four engineered strains BMB8901-171, BMB8902-171, BMB8901-4Q7 and BMB8902-4Q7. SDS-PAGE results indicated that all recombinant strains express 88 kD vegetative insecticidal protein. Bioassay also showed that the proteins of genes vip83 and vip14 both have certain toxicity to Lepidopteran insect larvae such as Heliochis armigera, Spodotera exigua and Plutella xylostella. While the toxicity of vip protein from four engineered strains to Plutella xylostellas are highest, whose LC50 value is 28.6, 31.6, 45.4 and 37.6 microL/mL respectively. This study will contributed to construct high efficacy and wide spectrum engineered strains on theory and reality.
Animals ; Bacillus thuringiensis ; genetics ; Bacterial Proteins ; chemistry ; genetics ; pharmacology ; Cloning, Molecular ; Insecticides ; pharmacology ; Pest Control, Biological ; Recombinant Proteins ; biosynthesis ; pharmacology

OBJECTIVETo assess the osseointegration capability of hydroxyapatite-coated porous titanium with bone morphogenetic protein-2 (BMP-2) and hyaluronic acid to repair defects in the distal femur metaphysis in rabbits.

METHODSPorous titanium implants were made by sintering titanium powder at high temperature, which were coated with hydroxyapatite by alkali and heat treatment and with BMP-2 combined with bone regeneration materials. And hyaluronic acid was further used as delivery system to prolong the effect of BMP-2. The implants were inserted into the metaphysis of the distal femur of rabbits. The animals were killed at 6, 12 and 24 weeks to accomplish histological and biomechanical analyses.

RESULTSAccording to the result of histological analysis, the osseointegration in BMP-2 group was better than that of the HA-coated porous titanium group. In push-out test, all the samples had bigger shear stress as time passed by. There was statistical difference between the two groups in 6 and 12 weeks but not in 24 weeks.

CONCLUSIONHydroxyapatite-coated porous titanium with BMP-2 and hyaluronic acid has a good effect in repairing defects of distal femur in rabbits, which is a fine biotechnology for future clinical application.

Animals ; Biomechanical Phenomena ; Bone Morphogenetic Protein 2 ; Bone Morphogenetic Proteins ; pharmacology ; Coated Materials, Biocompatible ; Durapatite ; Femur ; surgery ; Hyaluronic Acid ; pharmacology ; Osseointegration ; physiology ; Porosity ; Prostheses and Implants ; Rabbits ; Titanium ; Transforming Growth Factor beta ; pharmacology