Animals ; Heart Ventricles ; cytology ; Lipid Bilayers ; Potassium Channels ; Sarcolemma ; physiology ; Swine

In order to investigate the effects of phenylalanine, tyrosine and tyramine on the growth of Lycoris radiata suspension cells and the accumulation of alkaloids, the growth quantity of the cells as well as the content of alkaloids in cells were determined, which were treated with above three kinds of precursors alone and phenylalanine combined with tyrosine respectively. The results indicate that the addition of phenylalanine alone and addition of phenylalanine on the basis of tyrosine at high concentration (200 micromol/L) had no significant effect on the growth of Lycoris radiata suspension cells and the content of alkaloids in cells; whereas tyrosine and tyramine promoted the growth of the cells and alkaloids accumulation. Treated with tyrosine at high concentration (200 micromol/L), the content of alkaloids of the cells was 2.56-fold higher than that of the control group, the amounts of lycoramine (3.77 mg/g) and galanthamine (4.46 mg/g) were 6.61-fold and 6.97-fold higher than that of the control group, respectively. When treated with tyramine (200 micromol/L), the amount of alkaloids in Lycoris radiata suspension cells was 2.63-fold higher than that of the control group, and the amounts of lycoramine (4.45 mg/g) and galanthamine (5.14 mg/g) were 9.08-fold and 9.18-fold higher than that of the control group, respectively. The above results demonstrate that adding tyrosine and tyramine in the media significantly promoted the growth of the Lycoris radiata suspension cells and alkaloids accumulation in the cells.
Amaryllidaceae Alkaloids ; chemistry ; Cells, Cultured ; Culture Media ; chemistry ; Galantamine ; chemistry ; Lycoris ; chemistry ; growth & development ; Phenylalanine ; chemistry ; Plant Cells ; chemistry ; Plant Extracts

Objective:To clone and express the Staphylococcus aureus Efb(extracellular fibrinogen-binding protein) protein in Escherichia coli, to purify the expression product and prepare its functional antibody and to detect the functions of Efb protein for further studies on S.aureus infection.Methods: Efb gene was amplified by PCR using S.aureus NCTC-8325 genome DNA as template and cloned into the recombinant expression vectors pET28a. E.coli BL21(DE3) with the plasmid was induced with IPTG for protein production. The protein was purified by Ni~(2+) affinity chromatography. The function of Efb protein was determined by complement activity assay and inhibition ELISA.The polyclonal antibodies were prepared by immunizing the animals. Results: The purified recombinant Efb was obtained, which could inhibit the CH50 and AH50 effectively. The functional poly-antibodies of Efb were prepared.Conclusion:Efb could inhibit the classical pathway and alternative pathway of complement activation, and the antibodies against to Efb could block the inhibition of the classical pathway of complement activation induced by Efb.

BACKGROUND: This study was undertaken to investigate the expression of hypoxia-inducible factor-1α (HIF-1α) in rat cerebral cortex and the effects of β-sodium aescinate (SA) administration after return of spontaneous circulation (ROSC).METHODS: Sixty rats were divided into three groups: SA group, injected intraperitoneally with SA instantly after ROSC; control group, injected intraperitoneally with normal saline; and sham-operated group, without cardiac arrest or SA. The cardiac arrest model was established using asphyxiation and intravenous potassium chloride. Blood was sampled 1, 6, 12, and 24 hours after ROSC. Protein and mRNA levels of HIF-1α, VEGF and EPO were detected in the cerebral cortex by immunohistochemistry and real-time RT-PCR; serum levels of NSE and S100β were determined by enzyme-linked immunosorbent assays.RESULTS: Serum S100β and NSE were signifi cantly increased in the control group versus the sham-operated group 1, 6, 12 and 24 hours after ROSC (P<0.05). Protein and mRNA levels of HIF-1α, VEGF and EPO were signifi cantly increased in the control rats (P<0.05). Serum NSE and S100β were significantly decreased in the SA group versus the control group 1, 6, 12 and 24 hours after ROSC (P<0.05). Protein and mRNA levels of HIF-1α, VEGF and EPO were signifi cantly increased in the SA group (P<0.05).CONCLUSIONS: The expression of HIF-1α is increased in rat cerebral cortex after ROSC, and SA up-regulates the expression of HIF-1α. The up-regulation of HIF-1α improves the resistance of the cortex to ischemia and hypoxia and contributes to neuroprotection, possibly because of up-regulation of EPO and VEGF expression.

Objective To study the feasibility of rat models of renal edema based on disease combined with traditional Chinese medicine syndrome yin-deficiency and yang-deficiency types .Methods Gastric gavage of thyroxine and tail vein injection of Adriamycin were performed to induce a rat model of kidney Yin deficiency edema , and intramuscular injection of hydrocortisone and tail vein injection of Adriamycin were used to establish a rat model of kidney Yang deficiency edema .The 24 h urine protein content , triiodothyronine ( T3 ) , thyroxine ( T4 ) , cyclic adenosine monophosphate ( cAMP ) , cyclic guanosine monophosphate ( cGMP ) , estradiol ( E2 ) and testosterone ( T ) and other indicators were assayed to determine whether the rat models were successfully established .Result The rats of Yin deficiency edema group had clinical presentation such as hyperactivity , hair loss, dry stool, weight loss and temperature rise.Compared with the blank group , T3, T4, cAMP, E2, and 24 h urinary protein levels were significantly increased , cGMP and T content decreased , and cAMP/cGMP ratio was significantly increased , showing significant differences ( P<0.05 for all).The Yang deficiency edema animals displayed reduced activity , diarrhea, decreased body weight and body temperature, and other signs of disease.Compared with the blank group, T3, T4, cAMP, and E2 contents were significantly decreased , cGMP, T, and 24 h urinary protein levels were significantly increased , and cAMP/cGMP ratio decreased , showing significant differences ( P <0.05 for all ) .Conclusions The rat model of renal edema disease combined with TCM syndrome yin-deficiency was successfully induced by thyroxine in combination with Adriamycin , and the rat model of renal edema based on disease combined with TCM syndrome yang-deficiency is successfully established by administration of hydrocortisone plus Adriamycin .These two rat models demonstrate similar clinical manifestations of human renal edema based on disease combined with TCM syndrome yin-deficiency and yang-deficiency, respectively, therefore, may serve as useful tools for further research on this disease .

Objective To investigate the levels, phenotypes and functions of regulatory T cells (Treg) and Th17 cells in systemic sclerosis (SSc) patients and assess the role of imbalance between Treg and Th17 cells in the pathogenesis of SSc. Methods Peripheral blood mononuclear cells (PBMCs) of 31 SSc patients and 33 healthy controls were analyzed for the expression of CD4, CD25, CD45RA, the cytotoxic T-lymphocyte antigen-4 (CTLA-4), FoxP3, and IL-17 using flow cytometry. Treg immunosuppression capacities were measured in co-cultured experiments. The expressions of IL-17A, IL-22, IL-6 and IL-1β were measured by FlowcytoMix. The expression of FoxP3, CTLA-4, IL-17A, and RORC mRNA were measured by real-time polymerase chain reaction (PCR). The independent samples t-test was used to compare data between the groups. Results The frequency of Treg cells was significantly elevated in SSc [(3.6 ±1.1)%] compared with controls [(2.0±0.8)%, t=6.88, P<0.01)] and the expression of CTLA-4 was lower in Treg cells of SSc (P=0.034). The expression of CTLA-4 and FoxP3 mRNA was lower in SSc compared with controls (P<0.05), the immunosuppressive capacity of Treg cells were diminished in SSc (P=0.034). Th17 cells were increased in SSc (P<0.01); the levels of IL-17A, IL-22, IL-6 and IL-1β in the serums of SSc were higher than the controls (P<0.05). In SSc, CD25+FoxP3+IL-17+cells were increased[0.075%±0.032)%vs (0.049±0.027)%, t=3.52, P=0.029], but no difference of CD25+FoxP3+IL-17- was observed when compared with control. The expression of IL-17A and RORC mRNA were increased in SSc (P<0.01, respectively). Conclusion Both Treg and Th17 cells are increased in patients of SSc. Treg increases along with the deficiency of phenotype and function. FoxP3+IL-17+co-express cells are increased, and the balance between Treg and Th17 cells is broken.

Objective:To investigate the value of ADC derived from DWI combined with texture analysis derived from T 2WI fat suppressed images in distinguishing benign and malignant soft tissue tumors. Methods:The MRI and DWI images of 94 patients with soft tissue tumors (44 cases with malignant and 50 cases with benign) confirmed by pathology were analyzed retrospectively in the First Affiliated Hospital of USTC West District. ADC values of solid components were measured at GE ADW4.6 workstation. The texture features were extracted by manually drawing the ROI on the maximum level of the T 2WI fat suppressed images; the ADC values and texture parameters between the two groups were statistically analyzed by SPSS17.0, and the multivariate logistic regression model were conducted to analyze and calculate the diagnostic performance. Results:ADC value of benign and malignant soft tissue tumors was (1.6±0.3)×10 -3 mm 2/s, (1.2±0.5)×10 -3 mm 2/s, respectively, and the difference was statistically significant( t=-5.382, P<0.05). Taking 1.28×10 -3 mm 2/s as the critical value, the area under curve (AUC) for the diagnosis of benign and malignant soft tissue tumors was 0.783, the sensitivity was 92.00%, and the specificity was 65.91%. Among the texture features, the AUC of frequency size, skewness, Inertia All Direction_offset7, Inverse Difference Moment angle0_offset1, Inverse Difference Moment angle0_offset7 and Haralick Correlation All Direction_offset4_SD distinguishing benign and malignant soft tissue tumors were 0.825, 0.739, 0.826, 0.816, 0.820 and 0.783, respectively. The AUC, sensitivity and specificity of the best predictive model distinguishing benign and malignant soft tissue tumors were 0.930, 88.00% and 86.36% respectively using multivariate logistic regression analysis. Conclusion:ADC combined with texture analysis is of great value in preoperative differentiation of benign and malignant soft tissue tumors.

OBJECTIVETo observe the clinical therapeutic effect of acupuncture for treatment of reflux esophagitis of heat stagnation of liver and stomach type.

METHODSSixty-one cases were randomly divided into an acupuncture group (31 cases) and a medication group (30 cases). The acupuncture group was treated with needles at Zusanli (ST 36), Zhongwan (CV 12), Weishu (BL 21) and Neiguan (PC 6) mainly, once a day; and the medication group was treated with oral administration of 20 mg Omeprazole, once a day. The scores of clincial symptoms, comprehensive therapeutic effect, results of gastroscopy and pathology as well as recurrence rate etc. were observed before and after treatment.

RESULTSAfter treatment, the scores of symptoms significantly decreased in the two groups (both P < 0.01). The total effective rate of the acupuncture group was 90.3% (28/31), and 90.0% (27/30 )in the medication group, there was no statistical difference between two groups (P > 0.05); results of gastroscopy and esophageal mucosa pathology showed no statistical difference between two groups (both P > 0.05), the recurrence rate 12 weeks after treatment of 9.1% in the acupuncture group was lower than that of 42.9% in the medication grou p (P < 0.05).

CONCLUSIONAcupuncture has preferable short and long-term therapeutic effects for treatment of reflux esophagitis of heat stagnation of liver and stomach type.

Acupuncture Therapy ; Adolescent ; Adult ; Aged ; Esophagitis, Peptic ; pathology ; physiopathology ; therapy ; Female ; Humans ; Liver ; pathology ; physiopathology ; Male ; Middle Aged ; Stomach ; pathology ; physiopathology ; Young Adult

Objective To construct and apply a cell line screening Mycobacterium tuberculosis (Mtb)-specific tetramers of CD4+α/β T cell receptor(TCR). Methods The β chains of HLA class Ⅱ (DR) were amplified from tuberculosis patients by PCR. The pMT-HLA-DRB expression vectors that carries the HLA-DR 13 chain and pMT-HLA-DRA-P expression vectors which carries the genes of HLA-DR α chain loaded with Mtb antigen were transfected into S2 cells with the method of calcium phosphate transfection. The expressed Mtb peptide/HLA-DR complexes were primarily identified by the method of cell immunohistochemistry. The cell lines expressing Mtb peptide/HLA-DR complexes were used to screen tetramers of CD4+ TCR by flow cytometry. Results S2 cell lines expressing Mtb peptide/HLA-DR complexes on the cell surface were obtained, two kinds of Mtb specific tetramers of CD4+α/β TCR were screened. Conclusion S2 cell lines expressing Mtb peptide/HLA-DR complexes on the cell surface provide the solid basis of the further research on the TCR tetramers and are helpful for exploring new diagnostic study methods about tuberculosis and developing new vaccines.

BACKGROUNDCoronary artery lesions (CALs) are known to be the main complication in children with Kawasaki disease (KD). Instead of intravenous immunoglobulin (IVIG), corticosteroid therapy has been accepted to be used for children with KD who are unresponsive to IVIG. This study aimed to evaluate risk factors for CALs in children with KD.

METHODSWe retrospectively reviewed the clinical records of 2331 children with KD from January 2005 to December 2014. To identify the independent risk factors for CALs, multivariable logistic regression models were constructed using significant variables identified from univariate logistic regression analysis.

RESULTSThe incidence of CALs was 36.0% (840 of 2331), including 625 (26.8%) coronary artery dilations and 215 (9.2%) coronary artery aneurysms (CAAs). Multivariable logistic regression analysis identified that male, incomplete KD, longer fever duration, and C-reactive protein (CRP) >100 mg/L were independent risk factors for coronary artery dilatations. On the other hand, male, incomplete KD, longer fever duration, prolonged days of illness at the initial treatment, corticosteroid therapy, sodium ≤133 mmol/L, and albumin <35 g/L were the independent risk factors for CAAs. In addition, corticosteroid therapy, prolonged days of illness at the initial treatment, and albumin <35 g/L were the independent risk factors for giant CAAs.

CONCLUSIONSCALs might be associated with male sex, incomplete KD, longer fever duration, prolonged days of illness at the initial treatment, albumin <35 g/L, sodium ≤133 mmol/L, CRP >100 mg/L, and corticosteroid therapy. Corticosteroid therapy was an independent risk factor for CAAs and giant CAAs. Thus, corticosteroids should be used with caution in the treatment of KD with the risk for CALs.

Adolescent ; Adrenal Cortex Hormones ; adverse effects ; Child, Preschool ; Coronary Aneurysm ; chemically induced ; Female ; Humans ; Infant ; Logistic Models ; Male ; Mucocutaneous Lymph Node Syndrome ; drug therapy ; Retrospective Studies