Objective To investigate the role of mitogen-activated protein kinase(MAPK)signaling pathway regulating synthesis function of steroid hormone of porcine ovarian granulose cells.Methods Porcine ovarian granulose cells were cultured in medium added with dimethyl sulfoxide(DMSO)at concentration of 50μmol/L or with MAPK inhibitor PD98059 (PD) for 48 hours.Then,granulose cells in DMSO medium were added with activator of adenylate cyclase(20 μmoL/L)or blank agent for next 48 hours incubation,which were defined as observation group 1 and control group 1,similarly,granulose cells in PD medium were also added with activator of adenylate cyclase(20 μmol/L)or blank agent for next 48 hours,which were defined as observation group 2 and control group 2.The level of estradiol(E_2),progesterone (P)and testosterone (T) were detected by chemoluminescence.The mRNA expression of 17alpha-hydroxylase/17,20-lyase (CYP17) and aromatase P450 (P450 arom) were assessed by RT-PCR.Results (1)Hormone expression:the level of T,P and E_2 were(29.5±2.5)nmol/L,(80±5)nmol/L,(49±4) pmol/L in control group 1 and(42.3±3.4)nmol/L,(170±15)nmol/L,(75±6)pmol/L in control group 2,which showed significant difference(P<0.05).In the mean time,the level of T, P and E_2 were (106.2±7.6)nmol/L,(210±16)nmol/L,(130±11)pmol/L in observation group 2 and(47.2±3.5)nmol/L,(130±6)nmol/L,(81±6)pmol/L in observation group 1,which also reach statistical difference(P<0.05).(2)The expression of enzyme:the expression of CYP17 mRNA was inereased by 50% and P450 arom mRNA was decreased by 20% between control group 1 and 2. However, the mRNA expression of P450 arom and CYPI7 were upregulated remarkably, especially, the expression of CYP17 mRNA were increased by 125%. Conclusion MAPK signaling pathway plays an inhibitory role in regulating synthesis of steroid hormone of ovarian granulose cell.

Objective To study the local irritation of repeated intrathecal injection of Ziconotide Acetate,and to provide reference for irritancy evaluation ofintrathecal injection.Methods Sixteen New Zealand rabbits were assigned into two groupsat random:Control group and Ziconotide Acetate group,eight animals each group.Totally 50 μL saline or Ziconotide Acetate (100 μg/mL) were administrated by repeated lumbar intrathecal injection once daily for 7 d.Animal behavior was observed every day,and four animals in each group were sacrificed 2 d later after the last injection,the lumbar spinal cord was removed for histopathological examination and irritancy evaluation.The remaining animals were sacrificed for initancy evaluation 14 d later after the last injection.Results Only one animal died after anesthesia on day three in saline group,while no obvious adverse reactions were observed in other rabbits during the entire study,and no intrathecal irritant reactions of histopathological examination were found in both groups.The reversible minor mechanical damage was observed at the injection point,2 d after the last administration.Conclusion For 7 d repeated lumbar intrathecal injection in rabbits,no intrathecal irritant reactions observed in Ziconotide group,and the New Zealand rabbit could be used as a local irritation evaluation model.

【Objective】To investigate the molecular mechanism of dopamine (DA)-induced apoptosis in cultured cerebellar granule neurons (CGNs) and the effect of muscarinic cholinergic receptor (mAChR) agonist carbachol on it.【Methods】The apoptosis of neurons was measured by phase-contrast microscopy,Hoechst 33258 nucleus staining and DNA fragmentation agarose gel electrophoresis.The neuronal viability was measured by fluorescein diacetate (FDA) staining.The activation of extracellular signal-regulated protein kinase (ERK) was determined by Western blot.【Results】Dopamine increases the phosphorylation of ERK and induces apoptosis in CGNs,which is blocked by both carbachol and PD 98059.The protective effect and the inhibiting ERK phosphorylation of carbachol were blocked by atropine.【Conclusion】DA-induced apoptosis in CGNs may be mediated by activation of ERK.Carbachol protects CGNs from DA-induced apoptosis by activating mAChR and subsequent inhibition of activation of ERK.

Objective To study thyroid hormone changes in women with pre-eclampsia patients,the characteristics of thyroid disease and its relationship with pre-eclampsia.Methods From May 2011 to December 2012 171 patients with pre-eclampsia who delivered in Shengjing Hospital of China Medical University were recruited as prc-eclampsia(PE) group,among which 114 cases were defined as early onset pre-eclampsia (EP) group and 57 cases were defined as late onset pre-eclampsia (LP) group.And 171 healthy women with same age and same stage of pregnancy were selected as the control group.Their blood pressures were normal and they had no obstetrical complications.Serum thyrotropin (TSH),free triiodothyronine (FT3) and free thyroxine (FT4) levels were determined by solid-phase chemiluminescent enzyme immunoassay method (CMIA).Thyroid peroxidase antibody (TPOAb) and thyroglobulin antibody (TGAb) were measured by electro-chemiluminescent assay (ECLIA).The positive rate was calculated (TPOAb ＞ 5.6 U/L,TGAb ＞ 4.1 U/L were defined as positive result).The relationship between TSH,FT3,FT4 level and blood pressure was analyzed in women with pre-eclampsia.Results (1) The median values of TSH,FT4 and FT3 in PE group were 3.4 mU/L,(12.0 ± 3.0) pmol/L and(3.9 ± 0.9) pmol/L.In the control group,they were 1.9 mU/L,(13.4 ± 2.4) and (5.0 ± 1.3) pmol/L.There were statistically significant differences between the two groups(P ＜ 0.01).In EP group,the median values of TSH,FT4 and FT3 were 3.3 mU/L,(12.1 ± 3.4) pmol/L and (3.8 ± 0.9) pmol/L.The differences between EP group and the control group were statistically significant (P ＜ 0.01).In LP group,the median values of TSH,FT4 and FT3 were 3.4 mU/L,(11.9 ± 3.1) pmol/L and (3.9 ± 1.0) pmol/L.There were statistically significant differences compared to the control group(P ＜0.01).While there was no difference between EP group and LP group (P ＞ 0.05).(2) The positive rate of TPOAb and TGAb in PE group were 15.2％ (26/171)and 21.6％ (37/171),and were 12.3％ (21/171) and 14.6％ (25/171) in the control group.There was statistically significant difference in the TGAb positive rate (P ＜ 0.01),but the difference in TPOAb positive rate was not statistically different(P ＞0.05).The TPOAb positive rates in EP group and LP group were 12.3 ％ (14/114) and 21.1％ (12/57),respectively,with no statistically significant difference (P ＞ 0.05).And the positive rates of TGAb in EP group and LP group were 21.9％ (25/114)and 21.1％ (12/57),respectively,with no statistically significant difference(P ＞ 0.05).The positive rate of TPOAb in LP group and in the control group had statistically significant difference(P ＜0.01).(3) The morbidity of thyroid disease in PE group and in the control group were 47.4％ (81/171) and 16.4％ (28/171),with statistically significant difference (P ＜ 0.01).(4) The morbidity of subclinical hypothyroidism or hypothyroidism in PE group and in the control group were 45.0％ (77/171) and 16.4％ (28/171),with statistically significant difference(P ＜0.01).(5) The morbidity of subclinical hyperthyroidism in PE group and in the control group were 2.3 ％ (4/171) and 1.8 ％ (3/171),with no statistically significant difference (P＞0.05).(6) In PE group,women with TSH level of 0.3-3.3 mU/L had systolic pressure of(170 ± 21)mmHg (1mmHg =0.133 kPa)and diastolic pressure of(112 ± 15) mmHg; women with TSH ＞ 3.3 mU/L had systolic pressure of(166 ± 21)mmHg and diastolic pressure of(109 ± 13)mmHg.There was no statistically significant difference(P ＞ 0.05).But the diastolic pressure in EP group and LP group had statistically significant difference(P ＜ 0.01).In PE group,no correlation was found among TSH,FT4 levels and systolic pressure,diastolic pressure(P ＞ 0.05).FT3 level was negatively correlated to diastolic pressure (r =-0.172,P =0.023).Conclusions It is common that pre-eclampsia is complicated with thyroid dysfunction,mainly subclinical hypothyroidism.Thus it is nessesary to test thyroid hormone and thyroid antibodies in women with pre-eclampsia.The decrease of FT3 and FT4,the increase of TSH and the presence of TPOAb and TGAb are related with the presence of pre-eclampsia.

We isolated a high efficient antifungal strain A02 from forest soil in a suburb of Beijing. The result of polyphasic taxonomy confirmed that strain A02 belongs to Streptomyces lydicus. The fermented broth of the strain presented a stable and strong inhibiting activity against many plant pathogenic fungi. The purpose of this study was to ascertain the substance base of the antifungal activity of strain A02. We extracted the antifungal metabolite of A02 by using column chromatography with X-5 macroporous resin and 100-200 mesh silica gel respectively, and then purified it by LC-9101 recycling preparative HPLC with a SP-120-15 column (JAIGEL-ODS-AP). An active compound with purity over 99.845% was finally obtained. The chemical structure of the active compound was determined with spectroscopy methods, including ultraviolet spectrometry, infrared spectrometry, high resolution mass spectrometry and nuclear magnetic resonance. According to the analysis results, we identified the active compound as a tetraene macrolide antibiotic with the molecular weight of 665, the molecular formula C33H47No3 and the same chemical structure as natamycin. Our research revealed a new biosynthetic function for S. lydicus to produce natamycin, and an expanding application field for natamycin to be used for the control of fungal plant diseases.
Antifungal Agents ; chemistry ; isolation & purification ; Natamycin ; analogs & derivatives ; Soil Microbiology ; Streptomyces ; chemistry ; isolation & purification ; metabolism

Objeetive To compare the clinical value of early enteral nutrition (EEN) and combined enteral nutrition (EN) and parenteral nutrition (PN) in patients with hepatocellular carcinoma (HCC) after precise hepatectomy.Methods The prospective study was conducted.The clinical data of 60 HCC patients who underwent precise hepatectomy in the Affiliated Drum Tower Hospital of Nanjing University Medical School between January 2013 and December 2014 were collected.All patients were allocated into the EN group and combined EN+PN group by random number table method.Patients in the EN group and combined EN+PN group respectively received EEN and early combined EN+PN at 24 hours postoperatively for 7 days,total nutrient intakes between groups were consistent,with an average daily intake of nitrogen-feeding of 0.2 g/(kg · d) and an average daily intake of nonprotein calories of 25 kcal/(kg · d).Observation indicators:(1) comparison of nutritional indexes between groups;(2) comparison of liver function between groups;(3) comparison of postoperative conditions between groups.Measurement data with normal distribution were represented as-x±s.Comparisons at the same time between groups were respectively analyzed using the group-design t test.Comparisons of count data were analyzed using the chi-square test.Ordinal data were analyzed by the nonparametric test.Comparison of repeated measurement data were done using the repeated measures ANOVA.Results Sixty patients were screened for eligibility,and 30 in each group.(1) Comparison of nutritional indexes between groups:levels of albumin (Alb),globulin,prealbumin and hemoglobin (Hb) and lymph nodes count in the EN group were respectively (41±4)g/L,(28±4) g/L,(188±37) mg/L,(139± 17) g/L,(1.6±0.6) × 109/L before operation and (32±4) g/L,(23±5)g/L,(114±41)mg/L,(121±19)g/L,(1.1±0.7) ×109/L at 1 day postoperative1y and (34±5)g/L,(26±4) g/L,(169± 41) mg/L,(113 ± 16) g/L,(1.9 ± 1.1) × 109/L at 8 days postoperatively,with statistically significant differences in above indexes (F=2.23,4.45,8.96,5.21,2.18,P＜0.05).There were statistically significant differences in above indexes between pre-operation and 1 day postoperatively (t =9.79,8.53,9.81,4.56,5.77,P＜0.05) and between 1 and 8 days postoperatively (t =5.55,6.31,4.69,3.99,8.26,P＜0.05).There were statistically significant differences in levels of Alb,globulin,prealbumin and Hb between pre-operation and 8 days postoperatively (t=3.48,5.12,6.37,8.20,P＜0.05) and no statistically significant difference in lymph nodes count (t =2.48,P＞0.05).Levels of Alb,globulin,prealbumin and Hb and lymph nodes count in the combined EN+PN group were respectively (42±4)g/L,(28±6)g/L,(188±29)g/L,(142±16)g/L,(1.6±0.6)×109/L before operation and (31±5)g/L,(20±5) g/L,(96±31) g/L,(124± 16) g/L,(0.9±0.4) × 109/L at 1 day postoperatively and (34±4)g/L,(24±4)g/L,(143±18)g/L,(115±18)g/L,(1.4±0.7)×109/L at 8 days postoperatively,with statistically significant differences in above indexes (F=3.21,7.35,4.36,7.78,3.19,P＜0.05).There were statistically significant differences in above indexes between pre-operation and 1 day postoperatively (t =11.95,5.38,9.91,4.84,6.22,P＜0.05) and between 1 and 8 days postoperatively (t =9.45,7.66,4.98,3.15,4.79,P＜ 0.05).There were statistically significant differences in levels of Alb,globulin,prealbumin and Hb between pre-operation and 8 days postoperatively (t=9.31,8.44,6.57,5.25,P＜0.05).There were no statistically significant difference in lymph nodes count (t =3.11,P＞ 0.05) and in changing trends of Alb,globulin,prealbumin,Hb and lymph nodes count (F=8.54,3.83,7.21,6.33,3.91,P＞0.05).Levels of prealbumin at 8 days postoperatively were statistically different between groups (t =3.00,P＜0.05).(2) Comparison of liver function between groups:levels of alanine transaminase (ALT),aspartate transaminase (AST),alkaline phosphatase (ALP),glutamyl-transpeptidase (GGT),total bilirubin (TBil),direct bilirubin (DBil) and C-reactive protein (CRP) in the EN group were respectively (44±37)U/L,(44±30) U/L,(90± 29) U/L,(85 ± 49) U/L,(15.7± 5.4) μmol/L,(4.6± 2.3) μmol/L,(4.5 ± 3.2) mg/L before operation and (411±375)U/L,(379±323)U/L,(68±26)U/L,(55±49)U/L,(20.3±10.7)μmol/L,(7.8±5.5) μmol/L,(47.9± 30.6) mg/L at 1 day postoperatively and (107± 58) U/L,(43± 21) U/L,(100± 42) U/L,(111±71)U/L,(19.7±10.6) μmol/L,(8.8±5.9) μmol/L,(28.3±23.0) mg/L at 8 days postoperatively,with statistically significant differences in above indexes among pre-operation,1 and 8 days postoperatively (F=5.48,9.44,1.73,5.03,8.42,6.09,6.69,P＜0.05) and between pre-operation and 1 day postoperatively (t=5.12,5.36,5.35,0.45,2.88,3.82,6.95,P＜ 0.05).There were statistically significant differences in levels of ALT,TBil,DBil and CRP between pre-operation and 8 days postoperatively (t =6.71,4.14,5.96,6.33,P＜0.05) and in levels of ALT,AST,ALP,GGT and CRP between 1 and 8 days postoperatively (t =6.23,5.55,3.14,3.56,4.99,P＜0.05).There were no statistically significant difference in levels of AST,ALP and GGT (t =0.13,0.98,1.11,P＞0.05) and in levels of TBil and DBil (t =0.08,0.23,P＞0.05).Levels of ALT,AST,ALP,GGT,TBil,DBil and CRP in the combined EN+PN group were respectively (41±38) U/L,(43±25) U/L,(100±89) U/L,(106±46) U/L,(17.9±9.8) μmol/L,(6.2±3.6) μmol/L,(3.7±2.3)mg/L before operation and (462±409)U/L,(494±162)U/L,(73±53)U/L,(75±57)U/L,(28.1±18.8)μmol/L,(9.1±6.1) μmol/L,(40.7±26.2) mg/L at 1 day postoperatively and (90±47) U/L,(42± 16) U/L,(95±40) U/L,(110±66) U/L,(22.5± 14.5) μmol/L,(8.2±8.1) μmol/L,(26.9±24.6) mg/L at 8 days postoperatively,with statistically significant differences in above indexes among pre-operation,1 and 8 days postoperatively (F=5.96,3.73,6.94,3.88,7.97,4.14,5.25,P＜0.05) and between before operation and 1 day postoperatively (t=5.72,4.96,3.95,0.88,2.83,4.13,6.11,P＜0.05).There were statistically significant differences in levels of ALT and CRP between pre-operation and 8 days postoperatively (t =4.22,6.77,P＜0.05) and in levels of ALT,AST,ALP and GGT between 1 and 8 days postoperatively (t=7.01,5.21,4.38,6.15,P＜0.05).There were no statistically significant difference in levels of AST,ALP,GGT,TBil and DBil (t =0.29,1.65,1.92,0.33,P＞0.05) and in levels of TBil,DBil and CRP (t =0.09,0.37,0.58,P＞0.05).There was no statistically significant difference in changing trends of ALT,AST,ALP,GGT,TBil,DBil and CRP of 2 groups (F=7.18,2.23,3.94,5.88,4.72,2.17,6.53,P＞0.05) and in above indexes among pre-operation,1 and at 8 days postoperatively (t=0.27,0.42,0.91,0.12,0.96,0.24,0.59,0.32,0.49,1.27,0.88,0.07,1.07,1.45,0.23,1.11,0.88,0.32,0.35,0.93,0.21,P＞0.05).(3) Comparison of postoperative conditions between groups:times of initial defecation were respectively (3.2± 1.0) days and (4.2± 1.2) days in the EN group and combined EN +PN group,showing a statistically significant difference (t =3.21,P＜ 0.05).Conclusion EEN and combined EN+PN are safe and feasible in HCC patients after precise hepatectomy,and also can improve liver function recovery,but EEN has advantages of promoting protein synthesis and gastrointestinal function recovery.