OBJECTIVETo develop a new method for the determination of cholesteryl palmitate in Oviductus Ranae.

METHODA HPLC method was set up, using Zorbax Silica column and cyclohexane-diethyl ether (40:1) as mobile phase with a flow rate of 1.0 mL x min(-1), and the UV detection wavelength was 203 nm.

RESULTThe calibration curve was linear over the range of 0.60-8.92 microg (r = 0.9997), the average recovery of the method was 98.4%. RSD 1.8% (n = 6).

CONCLUSIONThe results showed that method was reliable and accurate.

Animals ; Cholesterol Esters ; analysis ; Chromatography, High Pressure Liquid ; methods ; Female ; Materia Medica ; analysis ; chemistry ; Oviducts ; chemistry ; Quality Control ; Rana temporaria

Nerve growth factor is one of the most important factors playing an important role in regulating the growth, development and survival of the neuron. The purified NGF from human placenta has been reported, the tissue from which can be isolated the NGF is very limited. It is important for basic research and clinic application to expression hNGF by genetic engineering. By polymerase chain reaction,gene fragment encoding the mature part of ?-NGF was amplified using the DNA of human placenta as template. The fragment was sequenced and inserted into expression vector pET-15b, and the recombinant expression vector pET15b-NGF was transformed into E.coli BL21(DE3)pLysS. After inducing with IPTG the NGF was higher expressed up to 25% of the total cell proteins. The expression product was purified with metal chelate affinity chromatography on Ni-IDA agarose under denaturing condition. The purity of rNGF was higher than 90% and yield of rNGF was 4.56mg/L expressing bacteria. SDS-PAGE revealed the NGF expression product had a Mr 16kDa. Western-blot displayed the recombinant product had strong immunological activity with rabbit anti-human ?-NGF polyclonic antibodies. The expression products were dealed with solubilizing inclusion bodies and refolding protein. The test of nerve fiber growth of chicken embryo DRG neurons displayed rNGF had biological activity.

OBJECTIVEStudies on the substance with nourishment Yin for a reasonable and rational auality appraise for turtleback.

METHODTo separate by chromatography and identifying with MS, 1H NMR, 13C NMR, IR.

RESULTTwo compounds were separated and identified as hexadecanoyl cholesterol ester and cholesterol.

CONCLUSIONThe two compounds are isolated from turtleback for the first time.

Animals ; Cholesterol ; isolation & purification ; Cholesterol Esters ; chemistry ; isolation & purification ; Materia Medica ; chemistry ; Turtles

OBJECTIVETo evaluate the practicability and validity of transcranial magnetic motor evoked potential monitoring (TMS-MEP) during spinal surgery.

METHODSFrom February 2001 to June 2004, 37 patients undergoing spinal surgery were involved, anaesthesia was maintained with volatile anesthetics in 11 operations and etomidate in 26. Analgesia was provided with fentanyl, and non-depolarizing muscle relaxant was given intermittently. MEPs elicited with transcranial magnetic stimulations were recorded from tibialis anterior muscles, simultaneously bispectral index (BIS) and train-of-four stimulation (TOF) were used to monitor the anesthesia depth and neuromuscular blockade respectively. The variety of MEP and its effect on surgical operation at different anesthesia depth and muscular relaxation were observed, and the muscle strength of the patients before and after operation were compared.

RESULTSThe 11 cases anesthetized with isoflurane or enflurane gave no response to TMS, the other 26 cases in which anaesthesia was maintained with etomidate and fentanyl gave satisfactory TMS-MEPs, but with significantly attenuated amplitudes and prolonged latencies (P < 0.05). Intraoperative MEP showed a grossly unchanged waveform, and its amplitude and latency had little fluctuation when anaesthesia and neuromuscular blockade maintained stable. When T(1) value of TOF at 40% - 60%, a steady MEP could be acquired and the muscular contraction after TMS should not interfere the operation.Seven of 26 cases had a MEP amplitude drop up to 50% or more during the operation, the surgical team was notified to avoid further spinal injury, at last only 1 case had a worsen muscle power after operation.

CONCLUSIONSMyogenic TMS-MEP is a valid and practicable technique for intraoperative monitoring, and the etomidate + fentanyl technique is adequate for its anesthesia. BIS and TOF monitoring are helpful to maintain the steadiness of the anesthesia and MEPs, which is very important for monitoring the changes of the MEPs.

Adolescent ; Adult ; Aged ; Anesthesia ; methods ; Evoked Potentials, Motor ; physiology ; Female ; Humans ; Male ; Middle Aged ; Monitoring, Intraoperative ; methods ; Spinal Cord ; surgery ; Spine ; surgery ; Transcranial Magnetic Stimulation ; Young Adult

OBJECTIVETo study the effects of Liangge San to the expression of CD14 and scaverger receptor(SR) in the kupffer cells of liver and the pathological changes of liver tissue of endotoxemia-mice.

METHODThe model was established with intravenous injection of lipopolysaccharide (LPS) and at the same time different dose Liangge San were given. The expression of CD14 and scaverger receptor were detected with immunohigtochemistry at the 2nd, 4th, 8th hour ofter injury and analyzed with computer image system, and the pathological changes of liver tissue were also observed.

RESULTAt the three different hours, the expression of CD14 and scaverger receptor in macrophages of liver of LPS-injury group showed significant increase and significant decrease respectively, compared with that of the blank-control group (P < 0.01). The expression in dexamethasone group and Liangge San different dose groups were intermediate between those in injury group and those in control group. Compared with expression of LPS-injury group, those of dexamethasone group and Liangge San different dose groups showed significant differences (P < 0.01), especially that of Liangge San high dose group. Liver cells showed vacuole change. Changes of CD14 and SR expression were paralleled with the severity of liver damages of the mice.

CONCLUSIONLiangge San can inhibite the up-regulation of CD14 expression and down-regulation of scaverger receptor expression in a dosage-dependent manner and also alleviate the damages of liver induced by LPS.

Animals ; Anti-Inflammatory Agents ; pharmacology ; Dexamethasone ; pharmacology ; Dose-Response Relationship, Drug ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Endotoxemia ; chemically induced ; metabolism ; pathology ; Female ; Kupffer Cells ; metabolism ; pathology ; Lipopolysaccharide Receptors ; metabolism ; Lipopolysaccharides ; Liver ; metabolism ; pathology ; Mice ; Plants, Medicinal ; chemistry ; Receptors, Scavenger ; metabolism

OBJECTIVETo investigate the association of -141C insert/delete polymorphism with schizophrenia in Wuhan of Hubei province.

METHODSA case-control study was conducted to analyze the polymorphism in the D(2) receptor gene promoter region with schizophrenia. A total of 120 cases of schizophrenia diagnosed according to CCMD-II R criteria and 100 normal controls were recruited in the study.

RESULTSIn this sample, the allele and genotype showed statistically significant differences between patients and normal controls (P<0.05).Especially, the frequency of -141C del was 11% in patients and 18% in control(OR 0.55, 95% CI 0.30-0.96; P<0.05). This allele was less common in schizophrenia than in normal controls (P<0.05).

CONCLUSIONThe -141C del polymorphism is associated with schizophrenia.The polymorphism may modify the association with other factors. Possibly -141C del in the DRD(2) promoter region is a strong candidate for a protective factor for this trait.

Adult ; Alleles ; Female ; Gene Frequency ; Genotype ; Humans ; Male ; Middle Aged ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Promoter Regions, Genetic ; genetics ; Receptors, Dopamine D2 ; genetics ; Schizophrenia ; diagnosis ; genetics

OBJECTIVETo find a way to discriminate operative reason from anaesthesia reason for the changes of intraoperative transcranial magnetic motor evoked potentials (MEPs).

METHODSIn 26 patients under Etomidate/Fentanyl anesthesia from February 2001 to June 2004, MEPs elicited by transcranial magnetic stimulation were recorded from tibialis anterior muscles, simultaneously bispectral index (BIS) and train-of-four stimulation (TOF) were used to monitor the anesthesia depth and neuromuscular blockade respectively. MEP, BIS and measurements of TOF at different anesthesia depth and muscular relaxation were recorded synchronously, statistical analysis of this data set was done in order to find the inherent relationship between these variables.

RESULTSUnder anesthesia, MEP amplitude was always positively correlated with the corresponding BIS and TOF value. A regression equation could be built, with which the MEP amplitude could be reckoned based on realtime BIS and T(1)/T(c). In case of spinal cord injury, the measured amplitude value would significantly deviate from predicted one, which suggested that the change of MEP was because of the operation, but not the anaesthesia or neuromuscular blockade. Each patient had his or her own regression equation, which was different from each other.

CONCLUSIONSThe establishment of regression equation from MEPs, BIS and TOF is very useful to distinguish reasons of the changes of transcranial magnetic MEPs during surgery, and with this technique, the intraoperative MEP monitoring should be more reliable and practicable.

Adolescent ; Adult ; Anesthesia, General ; Evoked Potentials, Motor ; physiology ; Female ; Humans ; Male ; Middle Aged ; Monitoring, Intraoperative ; methods ; Regression Analysis ; Transcranial Magnetic Stimulation ; Young Adult

OBJECTIVETo establish an animal model of tumor-associated depression and observe their biological behaviors and biochemical indices.

METHODSFour groups of SD rats kept in separate cages were subjected to tumor cell inoculation with or without chronic unpredictable moderate stress administered before or after the inoculation. The depressive behaviors of the rats were examined by open-field test, and the concentration of 5-HT in the hippocampus was measured by spectrophotofluorometry. The body weight of the rats and volume of the implanted tumor were monitored and sugar water test was performed.

RESULTSThe rats subjected to chronic stress displayed significant depression, manifested by reduction in movement in the central area and total movement distance with prolonged resting time and shortened time of activity. These rats maintained high levels of depression even 12 days after withdrawal of chronic stress. Compared with the control group, the depressive rats showed obviously reduced sugar water consumption and hippocampal 5-HT level. Tumors of different sizes were observed in all rats in the 4 groups.

CONCLUSIONA rat model of tumor-associated depression is established, and the tumor-bearing rats exhibit obvious depressive behaviors and reduced level of neural substance (5-HT), which provides a good basis for studying the association of depression with tumorigenesis,progression and prognosis of tumor.

Animals ; Carcinoma 256, Walker ; complications ; psychology ; Depressive Disorder ; etiology ; Disease Models, Animal ; Female ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Stress, Psychological ; complications

The chitosan/PHEA-blended hydrogels were prepared from PHEA and chitosan in various blend ratios. The water contents of the hydrogels were in the range of 50%-80% (wt). The attachment and growth of fibroblast cells(L929) on the hydrogels were studied. The results indicated the PHEA content in hydrogels has great effect on cell attachment but has little effect on the growth of L929 cells.
Animals ; Biocompatible Materials ; chemistry ; Cell Adhesion ; physiology ; Cell Division ; physiology ; Cells, Cultured ; Chitin ; analogs & derivatives ; chemistry ; Chitosan ; Fibroblasts ; cytology ; physiology ; Hydrogel, Polyethylene Glycol Dimethacrylate ; chemistry ; Mice ; Peptides ; chemistry ; Water ; chemistry

BACKGROUNDThere are few researches for the healing of metaphyseal fractures; moreover, the animal models to study the metaphyseal fractures are usually made by the oscillating saw osteotomy without reliable fixation, which is not in accordance with our current clinical practice. In this study, we established a new model to observe the healing process of metaphyseal fractures.

METHODSEighteen New Zealand rabbits were used in the study. The fracture model was created by splitting the medial tibial plateau in rabbits, then reset, and fixed with compression screws. At 1, 2, 3, 4, 6, and 8 weeks postoperatively, the tibial specimens were collected; firstly, a general observation and an X-ray examination of the specimens was done, and then they were embedded in methylmethacrylate and cut into sections with hard tissue slicer. The sections were stained with Giemsa reagent and examined under light microscopy.

RESULTSThere was no fracture displacement in the tibial specimens of all time points, except for one showing a collapse. No external callus formation could be observed by X-ray and general examination. After 1 week of the operation, the fracture gap was filled by mesenchymal tissue; 2 weeks postoperatively, a large number of woven bones were formed; from the third week onwards, the woven bone began to turn into lamellar bone, and new trabecular structure began to form. In all of the slices, no obvious chondrocytes formed in fracture areas; thus, there was no endochondral ossification.

CONCLUSIONSThis model was an ideal fracture animal model and suitable for the study of metaphyseal fracture healing. The X-ray and histological images demonstrated that metaphyseal fracture healing was a process of direct bone healing through intramembranous bone formation under the conditions of minor trauma, good reduction, and firm fixation.

Animals ; Fracture Healing ; physiology ; Fractures, Bone ; diagnostic imaging ; pathology ; Rabbits ; Radiography