OBJECTIVETo investigate the effect of removing bacterial endotoxin in the key processes of Reduning injection.

METHODThe content of bacterial endotoxins was detected by kenitic-turbidimetry and the removal efficacy was studied before and after using 0.8% of activated carbon and ultrafiltration with molecular weight cut-off of 10 x 10(3).

RESULTThe adsorption rate of bacterial endotoxins was 78.7% by using activated carbon, while the removal efficacy of bacterial endotoxins was 99.6% with ultrafiltration membrane at cut-off molecular weight 10 x 10(3).

CONCLUSIONThe key technology can effectively guarantee the safety of Reduning injection.

Adsorption ; Endotoxins ; isolation & purification ; Injections ; Pyrogens ; isolation & purification ; Ultrafiltration

Preclinical evaluation of medical devices (prototype products) offers the opportunity to investigate and study the intended use of device materials. Preclinical evaluation programs are designed to determine the efficacy, safety, and biocompatibility of biomaterials, prostheses, and medical devices. The purpose of safety testing is to determine if a material presents potential harm to the human; it evaluates the interaction of the material with the in vivo environment and determines the effect of the host on the implant. Preclinical evaluation is the determination of the ability of the prototype product to perform with appropriate host response in a specific application, considered from the perspective of human clinical use. Therefore, preclinical data should include materials science and engineering, biology, biochemistry, medicine, host reactions and their evaluation, the testing of biomaterials, and the degradation of materials in a biological environment.
Animal ; Carcinogenicity Tests ; Equipment and Supplies*/adverse effects ; Hemolysis ; Human ; Pyrogens/toxicity ; Sterilization

Endotoxin removal is essential for the safety of biological products. To remove endotoxin efficiently, we used polymyxin B (PMB) affinity adsorbent to remove endotoxin from protein solutions by static adsorption. We studied the effects of spacer length and ligand density of the affinity adsorbent, pH, salt type and concentration, protein type and concentration, endotoxin concentration, and additive on endotoxin removal and protein recovery. Endotoxin content and protein concentration were determined by test and Lowry assay respectively. The results showed that PMB affinity adsorbent had high capacity, high adsorption speed, high removal efficiency and good reusability. In addition, ligand density, pH, salt concentration and the isoelectric point and hydrophobicity of protein all had remarkable influence on the endotoxin removal. Under the optimal conditions, the recoveries of hemoglobin, human serum albumin and lysozyme were 87.2%, 73.4% and 97.3%, respectively, and the corresponding endotoxin removal rates 99.8%, 97.9% and 99.7%, respectively. This study illustrated the effects of solution conditions on the efficiency of endotoxin removal and protein recovery, and would provide useful reference for the efficient removal of endotoxin from biological products.
Adsorption ; Chromatography, Affinity ; methods ; Drug Contamination ; prevention & control ; Endotoxins ; isolation & purification ; Hemoglobins ; isolation & purification ; Polymyxin B ; chemistry ; Proteins ; isolation & purification ; Pyrogens ; isolation & purification ; Serum Albumin ; isolation & purification ; Solutions

AIMTo investigate the possible central mechanism of antipyretic effects of Chinese medicine gypsum.

METHODSGypsum was injected after the fever model was established. The firing rate of thermosensitive neurons in preoptic-anterior hypothalamus(PO/AH) region was recorded by using extracellular microelectrode technique.

RESULTSThe injection of pyrogen evoked decrease in firing rate of the warm-sensitive neurons and increase in the cold-sensitive neurons in the region of PO/AH; the changes of the firing rate of pyrogen- treated warm-sensitive and cold-sensitive neurons could be reversed by the injection of gypsum.

CONCLUSIONThe result may suggest that antipyretic action of gypsum is mediated by its influences on the thermosensitivity neurons in the region of PO/AH.

Action Potentials ; Animals ; Antipyretics ; pharmacology ; Calcium Sulfate ; pharmacology ; Cats ; Fever ; physiopathology ; Hypothalamus, Anterior ; physiopathology ; Male ; Materia Medica ; pharmacology ; Neurons ; physiology ; Preoptic Area ; physiopathology ; Pyrogens

To detect the toxic reaction degree for sheep acellular dermal matrix (ADM) in vivo or vitro by using hemolytic, pyrogen and cell-cytotoxic reaction experiments, respectively.
 Methods: Leach liquor of cross-linked and non-cross-linked sheep ADMs were set for cross-linked group and non-cross-linked group, respectively, with a positive control group (10 mL sterile water for injection in test tube) and a negative control group (10 mL 0.9% sodium chloride solution in test tube). The supernatants were obtained from each group and were measured for the absorbance. The hemolysis degree was calculated; 16 New-Zealand rabbits were selected and then divided into 4 groups, A, B, C and D group. The leach liquor of cross-linked and non-cross-linked sheep ADMs were injected into bodies of the 6 New-Zealand rabbits in the A and B groups, and then the body temperatures were measured in every half hour after injection, 6 times in total. The value of highest temperature among 6 measurements minus the normal temperature was the fever degree for the body temperature. Based on these fever degree, the criterion of biological pyrogen reaction for sheep ADM pyrogen experiment was evaluated; the mice fibroblasts were collected during logarithmic phase and were cultured in the nutrient medium containing sheep ADM leach liquor with different density. The absorbance was measured to evaluate relative growth rate for fibroblast.
 Results: The hemolysis degree for the group A and B are less than 5%. The summary of fever degree for New-Zealand rabbits were lower than 1.8 ℃. MTT experiment showed that the toxicity of 10%-90% or 100% leach liquor nutrient medium with sheep ADM for the mice fibroblast is at level 1 or level 2. There was no significant difference between leach liquor of cross-linked and non-cross-linked sheep ADMs (P>0.05). The effects on relative growth rate for mice fibroblasts were minor. 
 Conclusion: The hemolytic and pyrogen reactions for the sheep ADMs embedded in New-Zealand rabbit were within the evaluation criterion, and the effects on vitality and growth rate for the fibroblast were not significant.
Acellular Dermis ; adverse effects ; Animals ; Cell Culture Techniques ; Culture Media ; adverse effects ; toxicity ; Fibroblasts ; drug effects ; Growth Inhibitors ; pharmacology ; Hemolysis ; drug effects ; In Vitro Techniques ; Mice ; Pyrogens ; pharmacology ; Rabbits ; Sheep

The precise mechanism of set-point regulation in hypothalamus was not elucidated. Nitric oxide synthases (NOS) were detected in hypothalamus, however, the roles of NO in hypothalamus was not fully studied. So, we tested the effects of NO on body temperature because preoptic-anterior hypothalamus was known as the presumptive primary fever-producing site. NO donor sodium nitroprusside (SNP, 4 nmol, i.c.v.) elicited marked febrile response, and this febrile response was completely blocked by indomethacin (a cyclooxygenase inhibitor). But, ODQ (selective guanylate cyclase inhibitor, 50 microgram, i.c.v.) did not inhibit fever induced by SNP. The cyclic GMP analogue dibutyryl-cGMP (100 microgram, i.c.v.) induced significant pyreses, which is blocked by indomethacin. NG-nitro-L-arginine methyl ester (L-NAME, non selective NOS inhibitor) inhibited fever induced by interleukin-1 beta (IL-1 beta, 10 ng, i.c.v.), one of endogenous pyrogens. These results indicate that NO may have an important role, not related to stimulation of soluble guanylate cyclase, in the signal pathway of thermoregulation in hypothalamus.
Body Temperature ; Body Temperature Regulation ; Central Nervous System* ; Cyclic GMP ; Fever ; Guanylate Cyclase ; Humans ; Hypothalamus ; Indomethacin ; Interleukin-1beta ; NG-Nitroarginine Methyl Ester ; Nitric Oxide* ; Nitroprusside ; Prostaglandin-Endoperoxide Synthases ; Pyrogens ; Signal Transduction ; Tissue Donors