Aim To investigate the effect of the GABAA receptor on the analgesia,hypnosis and amnesia induced by GABA in mice.Methods GABA was intracerebroventricularly(icv) injected and securinine,a GABAA receptor antagonist,was intravenously(iv) injected after the GABA intracerebroventricularly injection.Then four techniques including hot-plate test,acetic acid-induced writhing test,awaken test as well as step-through test & step-down test were employed to evaluate the effects on the pain index in hot-plate test(HPPI),the writhing times,the sleeping time(ST) and the latency & the number of errors.Results GABA increased the HPPI and ST,but decreased writhing times and the latency & increased the number of errors in conscious mice;the above effects are dose-dependent.Securinine could antagonize the effects stimulated by GABA.Conclusion GABA lead to analgesia,hypnosis and amnesia.GABAA receptor might be the important target for these effects.

Objective: To compare the efficacy of moxibustion with different doses for knee osteoarthritis (KOA), and explore the correlation between moxibustion dose and clinical efficacy. Methods: Sixty-eight patients with KOA who met the inclusion criteria were randomly divided into a 20-minute moxibustion group and a 40-minute moxibustion group by the random number table method, with 34 cases in each group. Dubi (ST 35), Neixiyan (EX-LE 4) and Heding (EX-LE 2) were used for moxibustion in the two groups. Each treatment lasted 20 min or 40 min for each point in the 20-minute moxibustion group and 40-minute moxibustion group, separately; the treatment was given 3 times a week and lasted for 4 weeks. The visual analog scale (VAS), Western Ontario and McMaster University osteoarthritis index (WOMAC) and traditional Chinese medicine (TCM) symptom scores were evaluated before and after treatment to compare the efficacy between different moxibustion doses for KOA. Results: After treatment, the total effective rate was 87.5％ in the 40-minute moxibustion group, versus 70.0％ in the 20-minute moxibustion group, and the difference in the total effective rate between the two groups was statistically significant (P<0.05). After treatment, the VAS scores, the total WOMAC scores and the component scores of pain, stiffness and dysfunction, and the TCM symptom scores in both groups all changed significantly when compared with those before treatment (all P<0.05). After treatment, the between-group differences in the VAS score, the total WOMAC score and the component scores of pain and dysfunction, and the TCM symptom score were statistically significant (all P<0.05), while the difference in the stiffness score in WOMAC showed no statistical significance (P>0.05). Conclusion: Either 20-minute moxibustion or 40-minute moxibustion can relieve pain, improve stiffness, dysfunction, and TCM symptoms for KOA; and 40-minute moxibustion is better in relieving pain, improving dysfunction and TCM symptoms.

To obtain single chain variable fragment (scFv) and bivalent single chain variable fragment (bsFv) against transferrin receptor, up-stream and down-stream primers were designed according to the complementary sequences of FR1 region of variable heavy (VH) and FR4 of variable light (VL), respectively, which contained inter-linker G4S and the restriction endonuclease SfiI, AscI and NotI. Two pieces of scFv fragments were first amplified through PCR and then inserted into plasmid pAB1, which could express scFv protein once induced by IPTG in the host bacteria. To express scFv and bsFv, E. coli TG1 was cultured in LB broth and was induced by IPTG. The restriction enzyme digestion map and DNA sequencing demonstrated that scFv and bsFv genes were successfully inserted into the expression plasmid. SDS-PAGE and Western blotting revealed the protein band at 35kD and 60kD, which were consistent with the molecular weight of scFv and bsFv respectively. Flow cytometry showed that scFv and bsFv harbored the specific binding activity with TfR expressed in various tumor cells, and the avidity of bsFv was higher than that of the parent scFv.
Base Sequence ; Cloning, Molecular ; Escherichia coli/genetics ; Escherichia coli/metabolism ; Genetic Vectors/genetics ; Hep G2 Cells ; K562 Cells ; Molecular Sequence Data ; Receptors, Transferrin/*immunology ; Recombinant Fusion Proteins/biosynthesis ; Recombinant Fusion Proteins/genetics ; Single-Chain Antibodies/*biosynthesis ; Single-Chain Antibodies/genetics

Objective To explore the effect of age on neurogenesis of dentate gyrus granule cell and the impact on differentiation of newborn cells in rats.Methods SD rats were selected and divided into 5 groups according age of 7,14,28,60,180 d(n=8),and the neurogenesis of dentate gyrus granule cell in hippocampus with normal development was detected,using 5-bromo-BrdU(BrdU) labled newborn neuron and ?-tubulin protein(TuJ1) and glial fibrillary acidic protein(GFAP) labeled glial cells,and understood the newborn cells to neurons and glial cell differentiation ratio.Results Neurogenesis was found in dentate gyrus granule cell layer with hippocampus of all different age rats.Various forms of cells with a larger nucleus that were round,oval,diamond were distributed over the entire granule cell layer.BrdU-positive cells within each group were 158.07?5.37,141.28?7.27,116.93?9.24,76.56?6.88,41.42?4.45,the number of BrdU-positive cells were reduced with the growth of rats(P0.05);4%-5% newborn cells expressed GFAP.In addition,some of the BrdU-positive cells at the same time did not express TuJ1 or GFAP.Conclusions There are neurogenesis in dentate gyrus granule cell in rats of different age.The new born cells mostly differentzate into granule neuron cell.The capability of cell proliferation are decreased with the growth of age.

OBJECTIVETo observe the anti-inflammatory effect of total saponins of Panax japonicus on LPS-induced RAW264. 7 macrophages.

METHODThe effect of total saponins of P. japonicus of different concentrations on RAW264. 7 cell viability was determined with the MTT method. The NO kit assay was adopted to detect the NO release of total saponins of P. japonicus to LPS-induced RAW264. 7 cells. The enzyme linked immunosorbent assay (ELISA) was used to detect the secretion of tumor necrosis factor-alpha (TNF-alpha) and interleukin 1-beta (IL-1beta). The reverse transeriptase-polymerase chain reaction (RT-PCR) was used to determine the expression of inducible nitric oxide synthase (iNOS) ,TNF-alpha,IL-1beta. The protein expression of nuclear transcription factor-kappaB p65 (NF-kappaB p65) was tested by Western blot.

RESULTThe safe medication range of total saponins of P. japonicus was less than 80 mg x L(-1). Compared with the LPS model group, total saponins of P. japonicus high, middle and low dose groups (0.1, 1, 10, 40 mg x L(-1)) could significantly reduce the secretion of NO, TNF-alpha, IL-1beta of LPS-induced RAW264. 7 cells, and inhibit the expressions of iNOS, TNF-alpha and IL-1beta mRNA and the protein expression of NF-kappaB p65.

CONCLUSIONThis study preliminarily proves the protective effect of total saponins of P. japonicus on LPS-induced RAW264.7 macrophages. Its action mechanism may be related to NF-kappaB signal pathway.

Animals ; Anti-Inflammatory Agents ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Inflammation ; drug therapy ; genetics ; immunology ; Interleukin-1beta ; genetics ; immunology ; Lipopolysaccharides ; adverse effects ; Macrophages ; drug effects ; immunology ; Mice ; NF-kappa B ; genetics ; immunology ; Nitric Oxide ; immunology ; Nitric Oxide Synthase Type II ; genetics ; immunology ; Panax ; chemistry ; Protective Agents ; pharmacology ; Saponins ; pharmacology

BACKGROUND: The maximal problem of patient with serious hepatitis and surgical doctor is whether they can get donator and rational therapy timely. Looking for the suitable preoperative therapy method to enhance the success rate of operation and improve patient's prognosis is the focus of this domain.OBJECTIVE: To investigate the therapeutic effect of artificial liver support system (ALSS) combining with allotransplantation of the liver on patients with serious hepatitis.DESIGN: Retrospective case analysis. SETTING: Organ Transplantation Center, the Third Affiliated Hospital of Anhui Medical University.PARTICIPANTS: Five male patients with serious hepatitis who underwent allograft liver transplantation were selected from Organ Transplantation Center, the Third Affiliated Hospital of Anhui Medical University form June 2004 to May 2005. Their age ranged from 25 to 48 years. Inclusion criteria: The diagnosis was in accordance with phase standard established at the National Infectious Disease and Parasitology Academic Meeting in September 2000; all patients had signs of routine liver transplantation; their patients fiercely requested the operation.METHODS: Plasma exchange (PE) combined with continuous veno-venous hemofiltration (CVVH) technique was used in this study. Donor who supplied lives was from 20-38-year patients. All of them and their family agreed to donate their organ and signed the donate file before operation. All of 5 patients were used classical no-by-pass orthotopic liver transplantation (OLT). MAIN OUTCOME MEASURES: They were follow-up visited for 21-32 months for rechecking liver and kidney function,RESULTS: All of 5 patients' operation was succeeded. One continued coma postoperative and his serum creatinine and urea nitrogen raised up progressively and complicated by pulmonary infection 1 week after operation and died 2 weeks after operation although given medical treatment hemodialysis positively. The rest recovered well. All of them discharged one month after operation smoothly.CONCLUSION: Allotransplantation of the liver is an utilizable method to treat serious hepatitis. ALSS can be used as an effective method of supportive treatment preoperatively.

Aged ; Aged, 80 and over ; Humans ; Kidney Diseases ; diagnosis ; etiology ; Male ; Middle Aged ; Nephelometry and Turbidimetry ; Pneumoconiosis ; complications ; Pulmonary Disease, Chronic Obstructive ; complications ; Retinol-Binding Proteins ; urine

Objective To investigate the effects of dietary consumption of monounsaturated fatty acid (MUFA) on the control of type 2 diabetes mellitus. Methods One hundred and eighty-five patients with type 2 diabetes mellitus from a community in Shanghai were randomly divided into MUFA intervention group (MUFA group,n=125) and control group (n=60). The patients in MUFA group consumed tea oil enriched in MUFA for 3 months,while those in control group consumed normal oil. The serum glucose (fasting plasma glucose and 2-hour postprandial glucose),fasting insulin and blood lipid were examined before intervention and three months after intervention,and the parameters were compared within groups and between groups. Results The serum fasting glucose,fasting insulin,total cholesterol and low density lipoprotein-cholesterol 3 months after intervention were significantly lower than those before intervention in MUFA group (P0.05),while the serum fasting glucose,total cholesterol and low density lipoprotein-cholesterol in MUFA group were significantly lower than those in control group 3 months after intervention (P0.05). Conclusion Dietary consumption of MUFA can improve the glucose and lipid metabolism in patients with type 2 diabetes mellitus.

Anal Canal/surgery* ; Anorectal Malformations ; Constriction ; Fecal Incontinence ; Humans ; Rectal Prolapse

OBJECTIVETo establish a method for determining nucleosides (adenoside and guanoside) in Siweilingzhi Mixture by HPCE.

METHODAdenoside and guanoside were separated within 25 min using an 20 mmol.L-1 borate buffer with 30 mmol.L-1 SDS and 5% Ethanol (adjusted to pH 10.0 with sodium hydroxide solution), with an operation voltage of 10 kV, temperature of 20 degrees C and a hydrodynamic injection time of 15 s. Seperations were carried out in a fused-silica capillary 75 microns id x 57 cm (effective length 50 cm) with peak detection by direct UV at 254 nm.

RESULTRegression equation of adenoside and that of guanoside were Y = 0.0705 + 0.01707X (r = 0.9995) and Y = 0.0232 + 0.01864X (r = 0.9999) respectively. The average recovery rate was 99.22% (RSD = 3.66%) and 104.3% (RSD = 1.91%) respectively. Nine samples were determined with the method.

CONCLUSIONThe method is simple, rapid and accurate with good repeatability and it can be used to determine nucleosides.

Adenosine ; analysis ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; isolation & purification ; Electrophoresis, Capillary ; methods ; Fermentation ; Guanosine ; analysis ; Reishi ; chemistry