A mutant strain RZ13 with high lipase productivity was obtained through treating the Rhizopus sp. RXF12 by UV and microwave,and its activity was 2.62-fold of the original one. The high lipase productivity of the mutant strain could be inherited after repetitious subcultures.The optimal culture conditions of the RZ13 for producing lipase were studied.The lipase with highest activity (67.32U/ml) was obtained on the conditions of 25℃,pH 8.0,5 %(v/v) single spore suspension (1?107spore/ml) and 120 r/min for 84h,and stable under 40℃ and pH 8.5. In addition,the lipase was immobilized by adsorption onto Mg-Al hydrotalcite at 25℃ for 4 h by screening carriers and optimizing the immobilization. The results showed that the optimal reaction temperature and pH were 35~55℃ and 7.5~9.0,respectively,much wider than the free lipase.

OBJECTIVETo compare the clinical efficacy differences between acupoint catgut embedding and Kuntai capsule for perimenopausal syndrome, so as to provide an effective treatment method for perimenopausal syndrome.

METHODSThirty-three cases in the embedding group were treated with acupoint catgut embedding at back-shu points and front-mu points of liver, spleen and kidney combined with syndrome differentiation and disease differentiation, ten days per times; the Kuntai group was treated with oral administration of Kuntai capsule, 4 capsules each time, three times per day. The Kupperman index (KI) was observed in the two groups before treatment after 10 days, 30 days and 60 days of treatment, respectively; the efficacy was evaluated according to the ratio of KI.

RESULTSAfter the treatment, as treatment proceeded, the score of KI and ratio of KI were gradually reduced in two groups; the score of KI and ratio of KI in the embedding group after 10 days of treatment was lower than those in the Kuntai group (both P<0.05); after 10 days of treatment, the total effective rate was 36.4% (12/33) in the embedding group, which was superior to 3.0% (1/33) in the Kuntai group (P<0.05); however, after 30 days and 60 days of treatment, the differences of each index between two groups were not statistically significant (all P>0.05).

CONCLUSIONBoth the acupoint catgut embedding and Kuntai capsule could reduce the score of KI and improve clinical symptoms, and the acupoint catgut embedding has certain advantage on the early stage of treatment.

Acupuncture Points ; Acupuncture Therapy ; instrumentation ; Adult ; Catgut ; Female ; Humans ; Middle Aged ; Perimenopause ; physiology ; Treatment Outcome

Objective To explore the value of color doppler ultrasonography by bed side in the early diagnosis of HIE in full term neonates.Methods The changes of cerebral parenchymal and cerebral arterial blood stream parameter on 35 cases of neonates clinically diagnosed HIE of mild and moderate degree and 40 cases of normal newborns on the 24,48 and 72 hours after birth were observed by color doppler ultrasonography by bed side.Results 1.The cerebral parenchyma was even echo in normal newborns,but it was maldistributed and reinforced in mild asphyxia neonates and it was more serious in moderate degree.The echo of cerebral parenchyma in mild degree was near normal in 48 hours after birth,while the echo of cerebral parenchyma in moderate degree was still maldistributed and reinforced in 48 and 72 hours after birth.2.There was obvious changes in the cerebral arterial blood stream parameter and hemodynamics of the asphyxia newborns compared with normals.The systolic peak velocity(Vs)and end diastolic velocity(Vd)of the cerebral arteries in mild and moderate degree were obviously lower than that of control group in 24,48 hours after birth(Pa0.05).3.Resistance index(RI)of the cerebral arteries in mild and moderate degree were higher than that of control group in 24,48 hours after birth(Pa0.05).Conclusion Color doppler ultrasonography by bed side is a convenient,noninvasive method for diagnosing HIE.

OBJECTIVETo explore the anti-tumor mechanism of the combination of cisplatin with DC vaccine in tumor-bearing mice.

METHODSB16 melanoma cells were treated with cisplatin at the final concentration of 20 µg/ml in vitro for 24 h. The expression of HMGB1, Hsp70 and TGF-β were detected by Western blot. B16 tumor-bearing mouse models were generated. The therapeutic effect of the combination of cisplatin (100 µg/mouse i.p., for sequential 3 days) and intratumoral injection of DC cells (3×10(6)/mouse, twice with a 7-day interval) in the tumor-bearing mouse models was evaluated. Expression of MHC II, ICAM-1 and CD86 was analyzed by flow cytometry. The mice were sacrificed at 28 days after tumor cell inoculation. The tumors were removed and weighed, and tissue samples were taken for pathological examination. Tumor infiltrating lymphocytes (TIL) were isolated by discontinuous gradient centrifugation. The distribution of T-reg and CD8(+) T cells in the TIL was analyzed by flow cytometry, and the ratio of CD8(+) T/T-reg was determined. The activity of cytotoxic lymphocytes (CTL) was determined by microcytotoxicity assay.

RESULTSCisplatin enhanced both the B16 cell apoptosis and HMGB1 expression. After loading with cisplatin-treated cell lysate, the expression of MHC II, ICAM-1 and CD86 on DC cells were (47.5 ± 8.8)%, (35.5 ± 8.3)% and (36.2 ± 9.2)%, respectively. At 28 days after tumor cell inoculation, the tumor weight of the control group was (2.1 ± 0.6) g, that of the cisplatin group was (0.3 ± 0.2) g and that of cisplatin + DC vaccine group was (0.5 ± 0.2) g, showing a significant inhibition of tumor growth (P < 0.01). Furthermore, the CD8(+) T/T-reg ratio and CTL activity in TIL were also significantly enhanced in the tumor-bearing mice treated with cisplatin + DC vaccine. When the effector-to-target ratio was 20:1, 10:1 and 5:1, the CTL activity in the cisplatin + DC vaccine treated mice was (25.0 ± 5.0)%, (22.0 ± 6.0)% and (14.0 ± 4.0)%, respectively, significantly higher than (8.2 ± 3.6)%, (6.7 ± 1.8)% and (3.6 ± 1.9)%, respectively, in the control group (all P < 0.01).

CONCLUSIONCisplatin promotes the anti-tumor effect of DC vaccine by down-regulating T-reg cells and enhancing the CTL activity in tumors.

Animals ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; B7-2 Antigen ; metabolism ; CD8-Positive T-Lymphocytes ; pathology ; Cancer Vaccines ; pharmacology ; Cell Line, Tumor ; Cisplatin ; pharmacology ; Dendritic Cells ; immunology ; metabolism ; Female ; Genes, MHC Class II ; HMGB1 Protein ; metabolism ; Intercellular Adhesion Molecule-1 ; metabolism ; Melanoma, Experimental ; pathology ; Mice ; Mice, Inbred C57BL ; Neoplasm Transplantation ; T-Lymphocytes, Cytotoxic ; immunology ; T-Lymphocytes, Regulatory ; pathology ; Tumor Burden ; drug effects

Objective To investigate and compare the curative effect between delayed percutaneous coronary intervention (PCI) for patients with acute myocardial infarction presenting 12-24 hours from symptom onset and medical therapy on acute myocardial infarction patients presenting with ST-segment elevation (STEMI). Methods Using a prospective,open,parallel,controlled research approach,186 patients with STEMI were divided into delayed PCI group(n=89),which received PCI within 12-24 hours after STEMI and medical therapy group(n=97),which received medical therapy after STEMI. All patients were followed up 1-6 months with average follow-up (5.6 ± 1.4) months. Data of hospitalization period, the cardiac structures detected by echocardiography such as left atrial diameter (LAD), left ventricular diastolic diameter(LVDd),left ventricular ejection fraction LVEF,left ventricular fractional shortening(LVFS),composite end point events and major adverse cardiac events(MACE)were compared between the two groups.Results Compared with medical therapy group, the hospitalization cycle was significantly shorter in delayed PCI group. Data of the LAD and LVDd were significantly decreased,but LVEF and LVFS were increased in delayed PCI group compared with those of medical therapy group at 30 d and 6-month follow-up. The incidence of MACE and composite end point events were significantly less in delayed PCI group than those of medical therapy group (P<0.05). Conclusion Delayed PCI treatment can decrease the time of hospital stay and decrease the incidence rates of MACE and composite end point events,and improve left ventricular function and prognosis of patients.

OBJECTIVETo investigate the effects of oxidized low-density lipoprotein receptor 1 (LOX-1) on secretion of adhesive molecules mediated by ox-LDL in human umbilical endothelial cells (HUVECs).

METHODSHUVECs with different concentration of ox-LDL (0, 10, 20, 50, 100 microg/ml) were incubated for 24 h, or HUVECs were pretreated with 250 microg/ml poly (I) or 250 microg/ml carrageenan for 2 h and then incubated with 50 microg/ml ox-LDL for another 24 h. Expression of LOX-1 was determined by realtime RT-PCR and Western blot. mRNA and protein of ICAM-1, VCAM-1 and E-selectin were examined by RT-PCR and Western blot respectively.

RESULTSIncubation of HUVECs with ox-LDL (10-100 microg/ml) enhanced the expressions of LOX-1, ICAM-1 and E-selectin in a concentration-dependent manner (P < 0.01). On the contrary, ox-LDL did not affect the expression of VCAM-1 by HUVECs. The expression of LOX-1, ICAM-1 and E-selectin induced by ox-LDL were reduced in HUVECs pretreated with 250 microg/ml poly (I) or 250 microg/ml carrageenan for 2 h and then incubated with 50 microg/ml ox-LDL for 24 h. This showed that both poly (I) and carrageenan obviously decreased the expression of LOX-1, ICAM-1 and E-selectin induced by ox-LDL.

CONCLUSIONox-LDL may upregulate the expression of LOX-1, ICAM-1 and E-selectin, and LOX-1 blocker may partly inhibit this upregulation. The results suggest that the expression of inflammatory molecules induced by ox-LDL in HUVECs is mediated by LOX-1.

Cell Adhesion ; Cell Adhesion Molecules ; Cells, Cultured ; E-Selectin ; metabolism ; Endothelial Cells ; metabolism ; Endothelium, Vascular ; metabolism ; Humans ; Intercellular Adhesion Molecule-1 ; metabolism ; Lipoproteins, LDL ; biosynthesis ; RNA, Messenger ; metabolism ; Receptors, Oxidized LDL ; metabolism ; Scavenger Receptors, Class E ; metabolism ; Umbilical Veins ; cytology ; Vascular Cell Adhesion Molecule-1 ; metabolism

Objective To investigate the immunosuppressive effect ofcurcumin on xenogenic skin transplantation in mice.Methods The skin transplantation model was established by an operation,tergal skin flaps from BALB/c donor mice transplanted to the back of C57BL/6 recipient mice.Then recipient mice were divided into five groups at random,namely sham,model,curcumin (50 mg/kg),Cyclosporin A (10 mg/kg) and curcumin + Cyclosporin A (50 mg/kg + 5 mg/kg).All mice were ip administered once daily for 10 d.The survival days of skin graft were recorded in all groups.The interleukin-2 (IL-2) levels in plasma of all mice were determined by ELISA 4 and 8 d after the operation,respectively.Results The mean survival time of skin graft in curcumin,Cyclosporin A and curcumin + Cyclosporin A groups were 14.77,16.81 and 19.96 d,respectively,which showed significant.differences comparing with 12.10 d of model group (P ＜ 0.01).Combination of curcumin and Cyclosporin A administration showed a longer mean survival days than curcumin or Cyclosporin A group (P ＜ 0.05 or 0.01).The IL-2 levels in plasma of mice in curcumin,Cyclosporin A and curcumin + Cyclosporin A groups on postoperative day 4 were 3.68,2.05 and 2.70 ng/mL,respectively,which were significantly reduced than 4.76 ng/mL of model group (P ＜ 0.05 or 0.01).The IL-2 levels in plasma of mice in curcumin,Cyclosporin A and combination of curcumin and Cyclosporin A groups on postoperative day 8 were 4.06,2.11 and 2.95 ng/mL,respectively,which were significantly reduced than 5.85 ng/mL of model group (P ＜ 0.01).Conclusion Curcumin may have a good immunosuppressive effect on mice with xenogenic skin transplantation.

Objective To investigate the diagnostic value of pregnancy-associated plasma protein-A (PAPP-A) in acute coronary syndrome (ACS) patients. Methods Forty-nine cTnI-negative patients with coronary artery disease who were documented by angiography [31 cases with ACS,18 cases with stable angina (SAP)], and 28 healthy persons were selected as controls. PAPP-A and hs-CRP were analysed with enzyme-linked immunosorbent assay (ELISA). Results Circulating PAPP-A and ha-CRP levels were significandy higher in patients with ACS than those in patients with SAP and controls (P < 0.05). PAPP-A threshold value of 2.79 μg/ml identified patients who had ACS with a sensitivity of 81.0% and a specificity of 84.6%. PAPP-A levels were correlated with hs-CRP levels in patients with ACS (r = 0.418, P < 0.01). Conclusion PAPP-A is a strong candidate marker of ACS, especially to eTnl-negative patients.

OBJECTIVETo explore the pathogenesis and significance of neuroendocrine breast carcinoma by detecting chromogranin A (CgA) in human mammary tissues.

METHODEighty-nine cases of human mammary tissues were collected to detect CgA expression using immunohistochemistry.

RESULTNo CgA expression was detected in normal or hyperplastic tissues, but its expression was found in mammary carcinoma tissues at the rate of 16.7%. A significant difference in CgA expression was found between cancer tissues and non-cancer tissues, but not between the cancer tissues with different pathological grades.

CONCLUSIONThe pathogenesis of mammary neuroendocrine carcinoma may involve the micro-environmental factors that affect the differentiation of stem cells to give rise to immature cells, cell differentiation in other lineages or transdifferentiation. CgA may serve as an immunological parameter for this type of breast cancer in routine screening test.

Breast ; metabolism ; Breast Neoplasms ; metabolism ; Carcinoma, Neuroendocrine ; etiology ; metabolism ; Chromogranin A ; metabolism ; Female ; Humans

OBJECTIVETo study the expressions of survivin, PTEN and their relationships with tissue grade and pathology stage in prostatic carcinoma (PCa).

METHODSThe immunohistological staining was used to evaluated the expressions of survivin protein and PTEN in 43 case of prostatic carcinoma (PCa) and 5 cases of benign prostatic hyperplasia (BPH).

RESULTSThe positive rate of survivin protein was 81.40%. Expression of survivin protein in 5 case of BPH was negative. The positive rate of PTEN was 30.23%, and the higher the grade and the clinical stage of tumors were, the lower the expression of PTEN was, PTEN of 5 case of BPH was positive.

CONCLUSIONThe positively correlation was found between the abnormal expressions of survivin protein, PTEN and the biological behavior of prostatic carcinoma (PCa). Detection of survivin combined with PTEN is valuable for diagnosing PCa, and evaluating malignancy extent and prognosis.

Aged ; Humans ; Immunohistochemistry ; Inhibitor of Apoptosis Proteins ; Male ; Microtubule-Associated Proteins ; biosynthesis ; Middle Aged ; Neoplasm Proteins ; biosynthesis ; PTEN Phosphohydrolase ; biosynthesis ; Prognosis ; Prostatic Neoplasms ; metabolism ; pathology