OBJECTIVETo observe the effects of electroacupuncture (EA) at "Dazhui" (GV 14) on Wnt-β-catenin signal pathway in annulus fibrosus cells in intervertebral disc in rats with cervical spondylosis.

METHODSForty SD rats were randomized into a control group, a model group, an EA group and a medication group, 10 rats in each one. Rats in the control group were treated with sham operation, only incision on local skin; rats in the remaining groups were made into cervical spondylosis models. After model establishment, rats in the control group and model group received fixed treatment under identical condition; rats in the EA group were treated with EA at "Dazhui" (GV 14), 30 min per treatment; rats in the medication group were treated with intragastric administration of meloxicam tablets. Treatments were both given once a day, and 14 days were taken as one session; there was an interval of 2 days between two sessions, and totally two sessions were given. After the treatments, immunohistochemistry was applied to measure the expression of Wnt, glycogen synthase kinase-3β (GSK-3β) and Axin in annulus fibrosus cells; western blot was used to test the expression of P-β-catenin.

RESULTSIn the control group, there were more positive cells of Wnt, GSK-3β and Axin, which were intensively distributed, deeply colored, and strongly positive; In the model group, there were less positive cells of Wnt, GSK-3β and Axin, which were sparsely distributed and weakly positive. The expression of Wnt, GSK-3β, Axin and P-β-catenin in the model group was less than that in the control group (all P < 0.05); expression of Wnt, GSK-3β, Axin and P-β-catenin in the EA group and medication group was higher than that in the model group (all P < 0.05); expression of Wnt, GSK-3β, Axin and P-β-catenin was not significantly different between EA group and medication group (all P > 0.05).

CONCLUSIONEA could delay the degeneration of intervertebral disc, which may be related to EA inhibiting signal pathway of Wnt-β-catenin.

Acupuncture Points ; Animals ; Electroacupuncture ; Female ; Fibrosis ; Glycogen Synthase Kinase 3 ; genetics ; metabolism ; Humans ; Intervertebral Disc ; metabolism ; pathology ; Male ; Rats ; Spondylosis ; genetics ; metabolism ; pathology ; therapy ; Wnt Signaling Pathway ; beta Catenin ; genetics ; metabolism

OBJECTIVETo study the regulation of luteolin on spleen cells and sarcoma S180 cells in normal ICR mice.

METHODSSpleen cells and S180 cells were incubated with different concentrations of luteolin (50, 100, 200, and 400 μmol/L). The effect of luteolin on spleen cells and sarcoma S180 cells was determined by MTT assay. The apoptosis was detected using propidium iodide staining flow cytometry. Intracellular reactive oxygen species (ROS) was determined by flow cytometric analysis. Activities of free radicals scavenging were determined by hydroxyl radical and DPPH tests.

RESULTSCompared with the solvent control group, 200 and 400 μmol/L luteolin increased the spleen cells viability (P < 0.05). Luteolin at 100, 200, and 400 μmol/L decreased activities of S180 cells (P < 0.01). The proportion of sub-G1 phase spleen cells was reduced after treated with 200 and 400 μmol/L luteolin (P < 0.05). The proportion of sub-G1 phase S180 cells was elevated after treated with 200 and 400 μmol/L luteolin (P < 0.05). Compared with the solvent control group, levels of intracellular ROS in spleen cells of ICR mice all increased; levels of intracellular ROS in S180 cells all decreased after treated with 50, 100, 200, and 400 μmol/L luteolin (P < 0.05). Luteolin scavenged hydroxyl radical and DPPH in a dose dependent manner.

CONCLUSIONLuteolin had bilateral regulation on viability and apoptosis of spleen cells and S180 cells (promoting the viability of spleen cells, inhibiting apoptosis of spleen cells, inhibiting the viability of S180 cells, and promoting apoptosis of S180 cells), which was worth further study and exploration.

Animals ; Apoptosis ; Apoptosis Regulatory Proteins ; metabolism ; Cell Survival ; Luteolin ; metabolism ; Mice ; Mice, Inbred ICR ; Reactive Oxygen Species ; Sarcoma ; Spleen ; metabolism

OBJECTIVETo investigate osthole effect on femoral tissue resorption activity of rat in vitro.

METHODSSix SD rats weighted (80 ± 5) g were used to isolate and culture femoral tissue (diaphyses and metaphysis) in vitro. The cultured tissue were devided into control group, estradiol group and osthole group. The femoral tissue was treated with final concentration of 1 x 10(-5) mol/L osthole and 1 x 10(-8) mol/L estradiol culture in vitro at 48 hours after cultured. Tartrate-resistant acid phosphatase (StrACP) activity, glucose and Lactic acid content, StrACP, MCSF (Macrophage colony stimulating factor) and CTSK (Cathepsin K) mRNA was detected by Real-Time RT-PCR were detected.

RESULTSConcetration of Alkaline phosphatase activity were 2226 and 2498 in 1 x 10(-5) mol/L osthole and 1 x 10(-8) mol/L estradiol respectively. As compared with control group, the activity of StrACP of 1 x 10(-5) mol/L osthole and 1 x 10(-8) mol/L estradiol were inhibited at 6, 9, 12 days (P < 0.05); under treatment of in l x 10(-5) mol/L osthole, the content of Lactic acid were increased and the content of glucose were decreased at 3, 6, 9 days (P < 0.05); StrACP, MCSF and CTSK mRNA expression level were inhibited at 6, 9 days (P < 0.05).

CONCLUSIONOsthole can inhibit bone resorption and raise the level of nutrition metabolism of femurs tissue.

Acid Phosphatase ; metabolism ; Animals ; Bone Resorption ; prevention & control ; Coumarins ; pharmacology ; Estradiol ; pharmacology ; Femur ; drug effects ; Glucose ; analysis ; Lactic Acid ; analysis ; Male ; Rats ; Rats, Sprague-Dawley

Objective To explore the effect of different proportions of mixed blood exchange transfusion on blood circulation in neonates with hemolytic disease.Methods Thirty-one newborn infants with hemolytic disease were treated by peripheral arteriovenous synchronization of exchange transfusion with different proportions mixed blood.AB type plasma was mixed with O type red blood cell(RBC) washing.The proportion for the treatment group was 1:1(the O type RBCs 2 U:the AB type plasma 200 mL),by exchange transfusion of haplotypes,in accordance with 80?mL/kg;the proportion for control group was 2:1(the O type RBC 4 U:the AB type plasma 200 mL),by exchange transfusion of double in accordance with 150-180 mL/kg.The indicators were detected,such as the exchange rate of neonatal serum bilirubin,RBC,hemoglobin(Hb),hematocrit(HCT),and the exchange transfusion quantity and days of hospitalization before and after the exchange transfusion were analyzed.Results The exchange rate of serum bilirubin of treatment group and control group was (44.92?3.99)% and (45.69?5.06)%,respectively,there was no significant difference between 2 groups(P=0.639),there was no significant difference of hospitalization days[(8.13?1.13) d vs(8.19?0.91) d]between 2 groups(P=0.884).After exchange transfusion in treatment group,the average level of the RBC,Hb and HCT were increased(P

Objective To detect the mutations of ATP2C1 gene in patients with Hailey-Hailey dis- ease (HHD).Methods PCR and direct sequencing were performed in 17 patients and 120 healthy controls to screen the mutations in the exons of ATP2C1 gene.Results Eight mutations were identified in nine probands, including three deletion mutations (nt1464-1487 del/nt1462-1485del,1523delAT,2375delTTGT),three splice site mutations (360—2A→G,1415—2A→T,2243+2T→C) and two missence mutations (C920T and G1942T).None of the above mutations was found in the controls.Conclusion Eight specific novel mutations were identified in nine probands of HHD,which could be causative factors of the disease.

Coronary Vessel Anomalies ; diagnosis ; Electrocardiography ; Humans ; Infant ; Pulmonary Artery ; abnormalities

Enzymatic synthesis of monoglycerides by glycerolysis of oil and fats in microaqueous solvent-free media was investigated by using lipase from pseudomonus fluorescens (PFL). Initial eutectic point(IEP) was substituted for melt point of oil and fats in Critical Temperature Theory. By investigating the glycerolysis under different IEP, it is showed that there is a relationship between composition of the oils and the yield of monoglycerides: Y = -0.0006 X3 + 0.0592 X2 - 0.8909 X + 26.753(13% < X < 76.5%), here X is the contents(W/W) of saturated fatty acid residue (C16 + C18) in the oils, Y is the yield of monoglycerides at 40 degrees C. The optimum isothermal reaction conditions for a system which IEP is 40 degrees C are: 40 degrees C, 3%-4.5% (W/W) water in glycerol, dosage of lipase is 500 u/g oil when the mole ratio of glycerol to oil is 2.5:1. The highest yield of monoglycerides is 81.4% in 48 h by means of programming temperature reaction.
Glycerides ; metabolism ; Glycerol ; metabolism ; Kinetics ; Lipase ; metabolism ; Palm Oil ; Plant Oils ; metabolism ; Pseudomonas fluorescens ; enzymology ; Substrate Specificity ; Temperature

Enzymatic esterification by reacting caparic acid with glycerol in solvent-free system was studied. Lipases from Pseudomonas fluoresces(PFL), Mucor miehei(MML) and Candida antarictica(CAL) possessed good catalytic activity. The optimal reaction conditions to convert capric acid with CAL are: 60 degrees C, 20-100 u of CAL per gram capric acid, 12% (W/W) of initial water content in glycerol. CAL does not express its 1,3-specificity in final product. Mechanical fraying denatured CAL partly. 96.4% of catalytic activity of CAL recovered after 5 batches of reaction. Extraction with sodium carbonate solution can decrease acid value of product from 9.8 mg KOH/g to 0.68 mg KOH/g. Applying the enzymatic esterification in open system, under vacuum or dehydrating with molecular sieves all dehydrate effectively. Molar ratio of reactants does not influence the total conversion of capric acid but influences the yield of monoglyceride. With certain protocols, the reaction period could be shortened dramatically; conversion of capric acid reached 96.9% in 5 h.
Catalysis ; Decanoic Acids ; isolation & purification ; metabolism ; Glycerol ; isolation & purification ; metabolism ; Lipase ; metabolism

AIM: To observe thickness and morphological changes of bulbar conjunctiva pre- and post epidemic hemorrhagic conjunctivitis ( EHC ) therapy by optical coherence tomography ( OCT) .
METHODS: Observed morphological changes and measured the bulbar conjunctiva thicknesses of 29 cases (36 eyes) of incipient (1-2d) EHC patients, who were received and treated by department of ophthalmology, the Putuo Affiliated Hospital of Shanghai University of Traditional Chinese Medicine from May 2013 to December 2013, by OCT. Then measured the thickness again on 7, 14d after the therapy.
RESULTS: Among 29 patients (36 eyes), 7d after the EHC therapy, in 27 cured eyes, the full-thickness ( before 344. 00±59. 91μm, after 230. 19±22. 16μm, t=11. 75, P<0-01); epithelial thickness ( before 56. 52±6. 19μm, after 51. 37±5. 53μm, t=4. 61, P<0. 01); and stromal thickness (before 287. 11±60. 56μm, after 178. 81±20. 20μm, t=10. 69, P<0. 01) of patients' bulbar conjunctiva were thicker than values measured after therapy with significant difference. Significant difference was also found for full-thickness ( before 361. 39±65. 56μm, after 233. 44±22. 57μm, the difference was statistically significant, t=14. 45, P<0. 01);epithelial thickness ( before 55. 50±6. 72μm, after 46. 67±5-24μm, t=10. 06, P<0. 01) and stromal thickness ( before 305. 61±66. 02μm, after 186. 78±21. 82μm, t=13. 11, P<0-01 ) of patients' bulbar conjunctiva between values measured before and 14d after therapy.
CONCLUSION: The OCT is able to measure the thickness of bulbar conjunctiva in EHC patients. An significant increase was found in full, epithelial and stromal thickness of EHC patients' bulbar conjunctiva. With recovery from the disease, subepithelial fluid, interlaminar fluid and edema of the bulbar conjunctival stroma faded away firstly, which provide references for clinical therapies of the EHC.

Objective To conduct a microbiological and parasitological investigation of experimental minipigs in Guangdong province. Methods Four major experimental minipig production units in Guangdong province were included in this investigation. Samples were taken from a total of 154 pigs of 4 brreds, i. e. , Bama minipigs, Juema minipigs, Tibet minipigs and Wuzhishan minipigs. Pig fur, scales, serum, rectal swabs and feces samples were collected for detection of 20 pathogens. The data were analyzed and compared among the production units and breeds. Results Mixed infections were detected in all the four institutions. The infection rates of 7 pathogens were rather high: Streptococcus suis type 2 (50. 7%), Actinobacillus pleuropneumoniae (40. 3%), Mycoplasma hyopneumoniae (100%), Japanese encephalitis virus (41. 3%), porcine circovirus type 2 (74. 8%), porcine transmissible gastroenteritis virus (73. 8%),gastroenteritis virus (44. 7%). Porcine parvovirus (26. 0%), pseudorabies virus(15. 6%) and intestinal worms (3. 2%) were also detected in some animals. The immune qualified rates of classical swine fever virus (62. 8%) and foot-and-mouth disease virus (35. 8%) were rather low. The immune qualified rate of pseudorabies virus was as high as 98. 4%. Besides, Salmonella, Brucella, swine dysentery snake like spirochetes, dermatophytes, influenza virus. Toxoplasma gondii, ectoparasites, and coccidia were not detected. Conclusions The results of this investigation indicate that epidemiological quality control of pathogens in experimental minipigs and efforts to establish high grade minipig population in Guangdong province remain to be strengthened. Our study also provides a basis for revision of local and even national standards for experimental minipigs.