IL-12重组质粒联合骨髓间充质干细胞对MCF-7人乳腺癌细胞侵袭及裸鼠移植瘤生长的影响分析

目的探讨IL-12重组质粒联合骨髓间充质干细胞（BMSC）对MCF-7人乳腺癌细胞侵袭及裸鼠移植瘤生长的影响。 方法分离纯化培养BMSC原代细胞,制备BMSC条件培养基,分析BMSC条件培养基对MCF-7细胞株体外增殖和凋亡的影响;建立裸鼠MCF-7转移瘤模型,分为对照组、BMSC组、IL-12组以及联合组,各组每3 d进行一次瘤内注射,共注射3次,15 d后处死裸鼠,比较各组裸鼠肿瘤、脾脏重量。采用χ²检验、t检验以及方差分析进行统计学分析。 结果BMSC条件培养基能够抑制MCF-7细胞增殖,且随着作用时间的延长其抑制作用升高[24 h（13.42±1.93）%,48 h（16.83±1.77）%,72 h（20.21±2.01）,F = 6.271,P = 0.000]。BMSC条件培养基处理MCF-7细胞,可有效促进MCF-7细胞凋亡,且随着作用时间的延长细胞凋亡率升高[24 h（10.82±2.18）%,48 h（18.73±2.95）%,72 h（28.15±3.52）%,F = 9.215,P = 0.000]。与对照组裸鼠肿瘤体积[1 d（124.12±9.28）mm³,3 d（582.41±17.28） mm³,7 d（983.42±42.10） mm³,15 d（793.46±109.38） mm³]比较,BMSC组[1 d（132.61±12.41） mm³,3 d（378.46±23.08） mm³,7 d（542.61±58.49）mm³,15 d（329.48±47.51） mm³]、IL-12组[1 d（119.85±13.10） mm³,3 d（322.75±26.49） mm³,7 d（518.37±67.54）mm³,15 d（302.17±68.53） mm³]以及联合组[1 d（123.41±8.94）mm³,3 d（217.85±24.03）mm³,7 d（318.29±47.32） mm³,15 d（189.27±37.58）mm³]裸鼠肿瘤体积生长速度均减慢,差异具有统计学意义（F_1d=0.827,F_3d=37.583、F_7d=31.472、F_15d=15.372,P < 0.001）;处死各组裸鼠后裸鼠肿瘤质量比较,BMSC组[（529.42±98.74） mg]、IL-12组[（544.01±117.85）mg]以及联合组[（327.55±78.56） mg]均低于对照组[（877.42±120.11）mg],且联合组裸鼠肿瘤质量最低,差异具有统计学意义（F = 10.821,P < 0.001）;而裸鼠脾指数BMSC组（7.58±1.21）、IL-12组（7.63±1.09）以及联合组（10.03±1.52）均高于对照组（5.37±0.89）,联合组裸鼠脾质量最高,差异具有统计学意义（F = 13.411,P < 0.001）。 结论BMSC在体内外均具有抑制MCF-7肿瘤增殖作用,联合使用pcDNA6/IL-12对裸鼠MCF-7转移瘤抑制具有着明显的协同作用。     

国产西罗莫司与原研品在体内外对细胞影响的比较实验

目的探讨国产西罗莫司与原研品对移植宿主外周血中免疫细胞的影响效果。方法体外实验:人膀胱癌T24细胞体外培养,分别加入国产西罗莫司和原研品,CKK-8法检测并比较细胞增殖活性受抑制的情况。体内实验:建立小鼠异位心脏移植模型,设立对照无手术组(对照组)、移植无治疗组(Tx组)、移植+国产西罗莫司组(Tx+YXK组)、移植+原研品组(Tx+RAPA组)。观察移植心脏搏动情况,受者脾脏的流式细胞学检测,以及脾脏及移植物中免疫细胞浸润的病理检查。流式细胞检测树突状细胞(DC),CD8+细胞和调节性T细胞(Treg),病理组织学检测及免疫组化染色比较两组免疫细胞浸润情况。两组间比较采用独立样本t检验,多组间比较采用单因素方差分析,两两比较采用LSD-t检验。结果体外实验结果显示,国产西罗莫司与原研品对T24细胞活力影响的差异无统计学意义(P>0.05)。体内实验结果显示,Tx组移植心脏于第7天停止搏动,Tx+YXK组和Tx+RAPA组在第10天心脏搏动仍有力、节律正常。(1)脾脏流式细胞检测显示,与对照组、Tx组比较,Tx+RAPA组、Tx+YXK组CD11c+I-A+CD86+DC细胞(15.88±4.73、22.90±3.86比4.51±1.57、5.40±2.54)、CD8+淋巴细胞数量(6.32±0.98、6.75±1.34比3.03±1.12、3.23±0.97)均降低,而Tx+RAPA组CD4+CD25+Foxp3+阳性细胞数量(15.06±3.42比7.87±1.95,10.88±2.08)升高(P均<0.05)。Tx+YXK组和Tx+RAPA组3种免疫细胞数量差异均无统计学意义(P>0.05)。(2)移植心脏病理免疫细胞组化染色灰度分析,Tx组、Tx+YXK组和Tx+RAPA组CD4,CD8,IDO和CD11b数量差异无统计学意义(P>0.05),与Tx组比较,Tx+RAPA组和Tx+YXK组CD11c(25143.52±3525.12比12936.30±766.94、14240.60±3124.67)、Foxp3阳性细胞浸润数量(500.78±238.33比46.05±68.16、49.22±25.82)降低(P均<0.05),Tx+YXK组和Tx+RAPA组比较差异无统计学意义(P>0.05)。(3)模型动物脾脏病理免疫细胞组化染色灰度分析,Tx组CD 4和CD8阳性细胞浸润数量较Tx+YXK组和Tx+RAPA组少,但差异无统计学意义(P>0.05),Tx+YXK组和Tx+RAPA组比较,各种细胞染色的IOD值差异均无统计学意义。结论使用国产西罗莫司与原研品两种药物后受者移植心脏和脾脏中的细胞浸润变化一致;在体外对细胞增殖、移植后抗排斥作用和体内免疫细胞的影响表现均一致。     

Effect of graft versus host reaction on cell cycle time in neonatal mouse jejunum

Abstract. The duration of cell cycle parameters in control mouse jejunum has been compared with that found following induction of a graft-versus-host reaction (GvHR) during the first 3 weeks of postnatal life.
Values for t_c , and t_G1 were found to decrease progressively during normal development: estimates for the whole crypt column in 21-day-old mice were approximately half to one quarter those found 6 days after birth 12.1 ± 0.5 hr and 24.2 ± 0.3 hr for t_c ; 2.8 ± 0.3 hr and 12.1 ± 0.3 hr for t_G1 respectively; (means ± SE). t_S and t_G2 were found to remain approximately constant during this period of neonatal development.
Injecting foreign spleen cells into 3-day-old mice produced no effect on crypt cell proliferation or cell cycle parameters measured 3 days later. GvHR mice studied 8 days after spleen cell injection, however, showed both an increase in crypt cell proliferation and decreases in the values for t_c and t_G1 , to levels similar to those normally found in 21-day-old control animals ( t_c 12.4 ± 0.4 hr and t_G1 5.4 ± 0.4 hr for 11-day-old GvHR mice). The possible mechanism leading to these changes is discussed.
The ability of GvHR to stimulate cell proliferation is used in the present work to test the hypothesis that the total number of cell divisions taking place after birth determines the temporal sequence of changes in disaccharidase content produced during neonatal development.     

Neuronal Regulation of Interleukin 6 Secretion in Murine Spleen: Adrenergic and Opioidergic Control

Abstract: The PNS was anticipated to be involved in the modulation of immune responses. To study aspects of this neuronal-immune communication, a recently developed tissue slice method was used to study the effects of adrenergic and opioidergic transmitters on interleukin 6 (IL-6) secretion in the spleen. The α₂-adrenergic agonist p -aminoclonidine (10⁻⁷ M ) inhibited IL-6 secretion (control vs. p -aminoclonidine, 100.0 ± 4.76 vs. 59.3 ± 6.6% of control values; p < 0.001). The α₁-adrenergic agonist methoxamine (10⁻⁸ M ) also inhibited IL-6 secretion (100.0 ± 4.8 vs. 71.5 ± 3.8%; p < 0.001). The endogenous opioids β-endorphin (10⁻¹⁰ M ), methionine-enkephalin (10⁻⁹ M ), and leucine-enkephalin (10⁻⁹ M ) inhibited IL-6 secretion as well ( p = 0.0051, p = 0.0337, and p = 0.0226, respectively). Electrical stimulation of spleen slices inhibited IL-6 secretion (100.0 ± 4.3 vs. 56.7 ± 4.6% of control values; p < 0.001). The involvement of α-adrenergic and opioidergic molecules in this electrically induced inhibition was shown by the use of antagonists. Electrical inhibition of IL-6 secretion was attenuated by phentolamine (10⁻⁷ M ; p = 0.0345), by naloxone (10⁻⁶ M ; p = 0.0046), by cyprodime (10⁻⁸ M ; p = 0.0014), and by the combination of cyprodime (10⁻⁷ M ) plus phentolamine (10⁻⁸ M ; p < 0.0001). We conclude from the complementary studies that the inhibition of IL-6 secretion induced by electrical pulses was mostly mediated by α-adrenergic and μ-opioidergic endogenous transmitters.     

Morphological and functional alterations induced in trout intestine by dietary cadmium and lead

Effects of oral administration of lead and cadmium on structure and function of trout intestine were investigated in Salmo gairdneri. Fish were fed either cadmium (5 mg kg*¹ fish per day) or lead (10 mg kg⁻¹ fish p er day) and sacrificed after a 15 or 30 day treatment.
Levels of cadmium and lead in kidney, liver and spleen were significantly increased by the 15th day of treatment.
Following both cadmium and lead treatment, morphological observations showed an increased mucous cell activity, a disruption of intestinal brush border and an increased renewal rate of absorptive cells.
Influx (J_ms), outflux (J_sm) of chlorine and sodium ion and net fluxes were measured on perfused intestinal segments and ouabain-sensitive sodium, potassium-ATPase (Na, K-ATPase) activity was determined on intestinal scrapings. Cadmium did not alter either sodium chlorine transepithelial fluxes or sodium, potassium-ATPase activity, but lead did. In the middle intestine, lead modified significantly transepithelial sodium and chlorine fluxes (J_ms Na: 3.21 ± 0.34 to 1.79 ± 0.29 and J_ms Cl: 3.32 ± 0.34 to 1.86 ± 0.22μmol h⁻¹ cm⁻², n = 6) after 30-day diet. J_net remain unchanged and Na, K-ATPase activity decreased. In the posterior intestine, lead altered only J_ms Na (1.95 ± 0.31 to 0.37 ± 0.09μmol h⁻¹ cm⁻²) and J_ms Cl. Consequently J_nes were also decreased.
So, although both cadmium and lead induce morphological disorders in the middle and in the posterior intestine of the trout, they have different effects on the absorption mechanisms of ions.     

THE f NUMBER OF PRIMARY TRANSPLANTED SPLENIC COLONY-FORMING CELLS

The proportion of murine haemopoietic stem cells that settled in the spleen, after transplanting spleen cells into lethally-irradiated recipient mice, was found by comparing the number of spleen colonies obtained by transplanting a whole spleen with an estimate of the total number of colony-forming units (CFU) present in the intact spleen. the latter number was estimated assuming that endogenous spleen colonies were produced from surviving spleen-derived CFU which exhibited the same survival parameters as transplanted CFU.
Account was taken of the post-irradiation loss of CFU from the spleen in the endogenous assay, which was found to be a reasonably constant factor for doses higher than about 100 rad X-rays.
The total measured number of CFU/spleen from transplantation was less than the number calculated in the intact spleen by a factor, the primary f number, of 0.03 ± 0.02.     

Changes of microcirculation in healthy volunteers and patients with septic shock in Xining

Objective: The purpose of this study is to investigate the characteristic of microcirculation in healthy volunteers and patients with septic shock in both Xining(2 260 m) and Nanjing(10 m). Methods: A total of 62 cases, 33 healthy volunteers, 22 cases in Xining,(2 260 m above sea level) and 11 cases in Nanjing(10 m above sea level); and 29 septic shock, 13 cases in Xining and 16 cases in Nanjing were collected. The total vessel density(TVD), perfused vessel density(PVD), proportion of perfused vessel(PPV) and microcirculation flow index(MFI) of both healthy volunteers and septic shock had been investigated by using sidestream dark field(SDF). Analyzed and managed the image data by using AVA3.0 software. Results: In the healthy volunteers in Xining area(22 cases),the volume of TVD(15.59 ± 2.58 mm/mm~2), PVD(15.58 ± 2.58 mm/mm~2) and PPV(96.60% ± 4.63%) were significant higher than the volume of TVD(10.0 ± 2.10 mm/mm~2), PVD(10.81 ± 2.38 mm/mm~2) and PPV(84.24% ± 8.00%) of the volunteers(11 cases) in Nanjing(11 cases). But the MFI(2.17 ± 0.31) of the healthy volunteers in the Xining was significant lower(P0.05) than the MFI(3.21 ± 0.34) in the healthy volunteers of Nanjing. In the septic shock group(13 cases) in the Xining, the volume of TVD(5.44 ± 1.94 mm/mm~2), PVD(4.18 ± 1.61 mm/mm~2), PPV(42.14%± 5.38%) and MFI(1.05 ± 0.32) compared with the volume of the healthy volunteers in Xining, the TVD(15.59 ± 2.58 mm/mm~2), PVD(5.58 ± 2.58 mm/mm~2), PPV(96.60% ± 4.63%) and MFI(2.17 ± 0.30) were significant lower(P0.05). In the healthy volunteers compare with septic shock group in Nanjing area, the TVD(6.80±1.72 vs 10.00±2.10, P0.05), PVD(5.86±1.58 vs10.81±2.38,P0.05), PPV(45.42±4.86 vs 84.24±4.86, P0.05), MFI(1.28±0.28 vs 3.21±0.34 P0.05), there was significant decreased. In the septic shock group in the Xining compared with the septic shock in Nanjing, there was no significant difference. 10 of 13 patients with septic shock were survived in Xining. 13 of 16 patients with septic shock were survived in Nanjing. Conclusion: The changes of physiological and pathophysiological characteristic in microcirculation induced by hypoxia would be useful for clinical treatment of septic shock at high altitude.     

Stress-Induced Sensitization of Dopamine and Norepinephrine Efflux in Medial Prefrontal Cortex of the Rat

Abstract: We examined whether prior exposure to chronic cold (17–28 days, 5°C) alters basal or stress-evoked (30-min tail shock) catecholamine release in medial prefrontal cortex, nucleus accumbens, and striatum, using in vivo microdialysis. Basal norepinephrine (NE) concentrations in medial prefrontal cortex did not differ between chronically cold-exposed rats and naive control rats (2.7 ± 0.3 vs. 2.5 ± 0.2 pg/20 µl, respectively). Basal dopamine (DA) efflux in any of the brain regions was not significantly different between chronically cold-exposed rats and naive rats. However, a trend for lower basal DA efflux in the cold-exposed relative to naive rats was observed in medial prefrontal cortex (1.5 ± 0.2 vs. 2.2 ± 0.3 pg/20 µl, respectively), nucleus accumbens (3.7 ± 0.8 vs. 5.4 ± 0.9 pg/20 µl, respectively), and striatum (4.4 ± 0.5 vs. 7.2 ± 1.5 pg/20 µl, respectively). In medial prefrontal cortex of rats previously exposed to cold, tail shock elicited a greater increase from baseline in both DA and NE efflux relative to that measured in naive rats (DA, 2.3 ± 0.3 vs. 1.2 ± 0.1 pg, respectively; NE, 3.8 ± 0.4 vs. 1.4 ± 0.2 pg, respectively). However, in nucleus accumbens or striatum of rats previously exposed to cold, the stress-induced increase in DA efflux was not significantly different from that of naive rats (nucleus accumbens, 1.8 ± 0.7 vs. 1.5 ± 0.3 pg, respectively; striatum, 1.9 ± 0.4 vs. 2.6 ± 0.7 pg, respectively). Thus, both cortical NE projections and cortically projecting DA neurons sensitize after chronic exposure to cold. In contrast, subcortical DA projections do not sensitize under these conditions.     

Ghrelin-GHSR通路在急性低氧暴露大鼠胃炎症反应中的调节作用

由胃合成分泌的食欲刺激激素（ghrelin）可通过结合并激活生长激素促分泌激素受体,（growth hormone secretagogue receptor,GHSR）在调节胃功能方面发挥重要作用。急性低氧暴露导致的消化系统营养吸收障碍和胃肠道炎症反应是否通过Ghrelin-GHSR通路调控尚无研究。本研究采用Wistar大鼠为研究对象,随机分为4组：低氧暴露0 h组、12 h组、24 h组和48 h组,低氧干预在10.2%氧浓度的低氧房中进行。干预前后记录体重;通过分子生物学检测指标评价胃组织炎症因子含量、食欲刺激激素和下丘脑GHSR mRNA相对含量和蛋白质表达水平。本研究证实,随着低氧暴露时间的延长,大鼠体重减少量逐渐增加（12 h：3.73±3.08 g、24 h：8.77±5.04 g、48 h：12.53±6.16 g）;胃组织炎症因子IL-2、IL-4、IL-10、TNFα和MCP-1蛋白含量在低氧12 h后增加（4816.9±983.7 / 9074.5±1107.8 / 18895.1±2967.5 / 37.1±9.8 / 143.5±12.5 pg/mL vs. 166.1±34.6 / 38.3±4.2 / 1429.6±123.9 / 1.7±0.3 / 13.5±2.1 pg/mL）,随着低氧时间延长,炎症因子水平逐渐下降至正常水平（24 h：846.4±94.8 / 1269.8±167.9 / 5769.7±892.6 / 7.5±2.1 / 39.3±8.5 pg/mL;48 h：546.5±97.3 / 374.9±84.9 / 1889.7±982.3 / 2.1±0.8 / 24.6±6.4 pg/mL）;低氧12 h后胃组织食欲刺激激素 mRNA水平较0 h组下降（0.49±0.06 vs. 1, P < 0.05）,48 h后上升（3.79±0.54 vs. 1, P < 0.01）,胃组织的食欲刺激激素蛋白含量在低氧24 h和48 h后均出现上升（1.23±0.15 / 1.16±0.12 vs. 1, P < 0.05）;下丘脑GHSR mRNA在低氧48 h后上升（1.99±0.29 vs. 1, P < 0.01）,蛋白质水平在低氧24 h和48 h后均出现下降（0.35±0.06 / 0.48±0.04 vs. 1, P < 0.05）。表明急性低氧暴露会导致Ghrelin-GHSR通路下调,进而促进胃组织炎症反应,而随着低氧暴露时长的延续,Ghrelin-GHSR通路可通过下调胃中炎症因子水平而避免消化系统的进一步损伤。     

Patients with Amyotrophic Lateral Sclerosis and Other Neurodegenerative Diseases Have Increased Levels of Neurofilament Protein in CSF

Abstract: In the present study we describe an ELISA to quantify the light subunit of the neurofilament triplet protein (NFL) in CSF. The method was validated by measuring CSF NFL concentrations in healthy individuals and in two well-characterized groups of patients with amyotrophic lateral sclerosis (ALS) and Alzheimer's disease (AD). The levels were increased in ALS (1,743 ± 1,661 ng/L; mean ± SD) and AD (346 ± 176 ng/L) compared with controls (138 ± 31 ng/L; p < 0.0001 for both). Within the ALS group, patients with lower motor neuron signs only had lower NFL levels (360 ± 237 ng/L) than those with signs of upper motor neuron disease (2,435 ± 1,633 ng/L) ( p < 0.05). In a second study patients with miscellaneous neurodegenerative diseases were investigated (vascular dementia, olivopontocerebellar atrophy, normal pressure hydrocephalus, cerebral infarctions, and multiple sclerosis), and the CSF NFL level was found to be increased (665 ± 385 ng/L; p < 0.0001). NFL is a main structural protein of axons, and we suggest that CSF NFL can be used to monitor neurodegeneration in general, but particularly in ALS with involvement of the pyramidal tract.     

类叶升麻苷通过上调miR-204对缺氧/复氧诱导H9C2细胞凋亡的抑制作用

目的探讨类叶升麻苷对缺氧/复氧（H/R）处理大鼠心肌细胞（H9C2）损伤的影响及其分子机制。 方法体外培养H9C2细胞,H/R （4 h/20 h）建立心肌细胞损伤模型,并采用1、10、100 μmol/L类叶升麻苷,转染miR-204模拟物阴性对照（miR-NC）、转染miR-204模拟物（miR-24）,100 μmol/L类叶升麻苷干预+转染miR-204抑制剂阴性对照、100 μmol/L类叶升麻苷+转染miR-204抑制剂干预H/R细胞。分别进行RT-qPCR、MTT、流式细胞术、Western blot检测miR-204表达水平、细胞活力、细胞凋亡率和相关蛋白表达,利用相应试剂盒检测乳酸脱氢酶（LDH）、丙二醛（MDA）、超氧化物歧化酶（SOD）和谷胱甘肽过氧化物酶（GSH-Px）水平,酶联免疫吸附试验（ELISA）检测白细胞介素-6 （IL-6）、白细胞介素-β （IL-β）和肿瘤坏死因子-α （TNF-α）的含量。两组间比较采用独立样本t检验,多组间比较采用单因素方差分析,组间两两比较采用SNK-q检验。 结果与心肌损伤模型比较,10、100 μmol/L类叶升麻苷的细胞凋亡率[（25.62±1.96）%比（18.17±1.27）%,（11.24±0.57）%]、Bax（0.71±0.05比0.51±0.04、0.29±0.03）、LDH [（243.16±11.31）比（121.22±4.52）,（94.39±2.82）U/g]、MDA [（1.82±0.07）比（1.13±0.04）,（0.92±0.04）nmol/mg]、IL-6 [（121.45±6.18）比（87.16±4.53）,（47.11± 2.24）pg/mL]、IL-1β [（229.82±8.48）比（175.32±8.73）,（113.14±5.63）pg/mL]和TNF-α表达水平[（138.18±6.60）比（92.24±4.04）,（61.53±4.17）pg/mL]降低,Bcl-2 （0.18±0.01比0.35± 0.03、0.52±0.04）、SOD [（18.72±1.26）比（38.81±1.51）,（45.43±1.29）U/mg]和GSH-Px表达水平[（58.74±2.28）比（89.24±2.82）,（94.66±3.05）U/mg]升高（P均< 0.05）;与miR-NC比较,转染miR-204的细胞凋亡率[（24.12±1.12）%比（9.26±0.49）%]、Bax （0.62±0.04比0.25±0.02）、LDH [（229.11±8.47）比（86.32±5.92）U/g]、MDA [（1.75±0.08）比（0.85± 0.05）nmol/mg]、IL-6 [（134.47±7.31）比（55.26±2.13）pg/mL]、IL-1β [（211.14±9.70）比（98.11±3.18）pg/mL]和TNF-α表达水平[（152.92±3.49）比（51.34±2.66）pg/mL]降低,Bcl-2 （0.22±0.01比0.57±0.03）、SOD [（20.92±1.38）比（47.68±1.76）U/mg]和GSH-Px表达水平[（62.65±2.76）比（91.13±3.80）U/mg]升高（P < 0.05）;10、100 μmol/L类叶升麻苷可提高H/R诱导H9C2细胞中miR-204的表达水平（P < 0.05）;且下调miR-204逆转了类叶升麻苷对H/R处理H9C2细胞凋亡、氧化应激和炎症因子的影响。 结论类叶升麻苷可能通过上调miR-204表达缓解H/R诱导的H9C2细胞损伤。     

δ-联蛋白通过AKT信号通路调控OGD/R后的炎症反应

δ-联蛋白(delta-catenin)作为高表达于神经系统的黏附蛋白质,在神经系统的功能发挥中有着至关重要的作用,但其在缺血缺氧性脑病中的研究尚未见报道。本文通过体外培养原代皮层神经元,构建氧糖剥夺/再灌注（oxygen-glucose deprivation/reperfusion, OGD/R）模型。用Western 印迹、LDH等方法显示,与对照组相比,δ-联蛋白在OGD/R模型后的不同时间点（0、4、12、24 和48 h）表达呈先降低后升高的趋势。在12 h表达量最低（0.48±0.08 vs 1.53±0.18,P<0.05）,在48 h表达升高到对照组水平（1.35±0.15 vs 1.53±0.18,P>0.05）。用siRNA慢病毒干扰δ-联蛋白表达后,ELISA结果显示,和对照组相比,OGD/R后,IL-1β和IL-18升高（24.80±1.64 vs 12.75±0.87,28.12±2.69 vs 12.99±1.24,P<0.05）,但在干扰δ-联蛋白表达后,和OGD组相比,IL-1β和IL-18释放降低（12.81±0.78 vs 24.80±1.64,14.27±1.37 vs 28.12±2.69,P<0.05）。Western 印迹结果显示,AKT信号通路磷酸化位点Ser 473活化增高（1.08±0.04 vs 0.85±0.06,P<0.05）,但Thr 308位点活化无改变（1.17±0.06 vs 1.11±0.08,P>0.05）。在siRNA慢病毒干扰并且联合使用AKT信号通路抑制剂GSK 690693后,和OGD+siRNA组相比,IL-1β和IL-18释放增高（24.58±0.99 vs 12.81±0.78,31.62±2.23 vs 14.27±1.37,P<0.05）。上述结果显示,δ 联蛋白通过AKT信号通路调控OGD/R后的炎症反应,这可作为δ-联蛋白功能研究的新的实验依据。     

龙须菜海藻场构建及其对水环境因子的影响

2010年2月～2010年5月,在汕头南澳岛海域,通过设计不同的主绳间距和苗绳间距研究了龙须菜(Gracilaria lemaneifomis)海藻场的构建模式及其对养殖附近海域环境的影响。结果表明,在不同的筏式养殖密度中,主绳间距在0.3～0.4m,苗绳间距在0.1～0.2m之间,龙须菜的生长速率较高。龙须菜海藻场构建后,养殖海域的无机氮(IN)平均值为133.40±15.18μg·L^-1,对照海域的无机氮平均值为153.81±11.97μg·L^-1,养殖海域显著低于对照海域(p<0.05);养殖海域的无机磷(IP)平均值为13.16±1.38μg·L^-1,对照海域的平均值为15.55±1.05μg·L^-1,养殖海域的显著低于对照海域的(p<0.05)。养殖海域叶绿素(Chl-a)的平均值为1.86±0.23mg·m^-3,对照海域的平均值为2.44±0.30mg·m^-3,养殖海域显著低于对照海域(p<0.05);养殖海域的透明度平均值为1.76±0.09m,对照海域的透明度为1.41±0.15m,养殖海域显著高于对照海域(p<0.05):养殖海域的溶解氧(DO)平均值为9.55±0.46mg·L^-1,对照海域的平均值为8.90±0.37mg·L^-1,养殖海域显著高于对照海域(p<0.05)。研究表明,构建龙须菜海藻场可以显著增加海水中DO浓度,抑制微藻生长繁殖,吸收水体营养盐,改善水质质量。     

丁酸通过增强肝线粒体功能缓解高脂诱导的小鼠肥胖

丁酸可以预防高脂日粮诱导的小鼠肥胖和胰岛素抵抗,但是否有治疗作用尚不清楚。本研究证明,高脂日粮诱导小鼠肥胖模型后,用80 mg/mL丁酸钠水溶液灌胃能够缓解肥胖。表观指标检测发现,丁酸钠显著降低肥胖小鼠的肝重（1.24 g ± 0.03 g 至1.08 g ± 0.04 g）、体重（32.46 g ± 0.50 g至28.35 g ± 0.58 g）和附睾脂重（1.33 g ± 0.13 g至0.81 g ± 0.08 g）及其与体重的比（4.06% ± 0.37%至2.83% ± 0.22%）。葡萄糖耐受实验和血液激素含量检测表明,丁酸钠部分缓解由高脂引起的葡萄糖不耐受,并显著降低血液中瘦素（3.71 ng/mL ± 0.62 ng/mL至1.50 ng/mL ± 0.26 ng/mL）和胰岛素（2.39 ng/mL ± 0.30 ng/mL至1.25 ng/mL ± 0.09 ng/mL）的水平。肝中脂质和糖原的生化检测表明,丁酸钠对肝中的甘油三酯、胆固醇和糖原的含量没有显著影响。通过RT-PCR实验发现,丁酸钠显著上调线粒体β氧化和解耦联相关的关键基因以及线粒体自身编码的8个基因的mRNA水平的表达,Western印迹检测表明,丁酸钠显著升高肝葡萄糖转运蛋白GLUT2和调控线粒体功能的关键蛋白PGC-1α的表达。上述结果提示,丁酸钠可能通过增强肝线粒体功能缓解食源性小鼠肥胖。     

Assay of bacterial and fungal activity in diesel contaminated soil using a 14C-glucose utilization method

Changes in the relative metabolism of soil bacteria and fungi following contamination with diesel were assessed using a modified substrate-induced respiration (SIR) method including selective antibiotic inhibition. ¹⁴CO₂ release from radiolabelled glucose was used as an indication of population activity. In a Sandy Gley Soil with no history of contamination, the population activity shifted from 38 ± 4% (bacterial): 62 ± 4% (fungal) to 73 ± 4% (bacterial): 27 ± 4% (fungal) after treatment with diesel.     

大蓟紫外光谱鉴别特征的初步研究

为了研究大蓟(Cirsium japonicum)紫外光谱的显著鉴别特征，以快速、准确地鉴别正品大蓟，采用多溶剂紫外光谱法分析福建省不同地区的自采大蓟和市售18批不同厂家商品大蓟在无水乙醇、氯仿及石油醚溶剂中紫外特征差异，并采用主峰值(含肩峰值)及主谷值的均值加方差归纳比较特征。结果表明，自采大蓟在无水乙醇溶剂中表现为“谷—峰—谷—峰”，即在273±3 nm、326±2 nm处的峰值和250±2 nm、298±1 nm处的谷值具有显著鉴别意义；在氯仿溶剂中，表现为“峰—谷—峰”，即在245±3 nm、273±4 nm处的峰值和260±3 nm处的谷值具有显著鉴别意义；在石油醚溶剂中表现为“肩峰—谷—峰”，即在233±3 nm处的肩峰、265±5 nm处的峰值和257±5 nm处的谷值具有显著鉴别意义。紫外光谱的峰谷特征可用于大蓟鉴别。     

Motility and fertilizing capacity of fresh and frozen-thawed spermatozoa in sturgeons Acipenser Baeri and A. ruthenus

The Siberian sturgeon ( Acipenser baeri ) and the sterlet ( A. ruthenus ) were injected with dried sturgeon pituitary (2 mg kg⁻¹), yielding 24 h later respectively 1.71 ± 0.5 and 1.65 ± 0.5 10¹¹ spermatozoa kg⁻¹ body weight. Spermatozoa were best activated with a solution of Tris HC1 50 mM, pH 8.0. The percentage of activated cells was 88 ± 4.4 in A. baeri (n = 5) and 68 ± 19 in A. ruthenus. (n = 5). In A. baeri , immediately after activation, the beat frequency of the flagellum and the mean velocity were in the range of 48–52 Hz and 100–300 μm⁻¹s, respectively. The beat frequency declined to 15 Hz at 30–40 and velocity to 100 μm s⁻¹ at 60 s post-activation. Only a small percentage of the spermatozoa remained motile after 3–4 min. In all cases spermatozoa showed mostly quasi-linear trajectories. Sperm was frozen in liquid nitrogen vapor immediately after dilution 1 v: 1 v in a cryopreservation medium (23.4 mM saccharose, 118 mM Tris-HCl pH 8.0, 20% egg yolk to which 15% DMSO were added). After fast-thawing procedure (25 s at 40°C), the percentage of motile spermatozoa (once activated in 118 mM Tris-HCl, pH 8.0) decreased to 23 ± 8.8 in A. baeri and to 15 ± 11 in A. ruthenus. The fertilizing capacity also decreased: 53 ± 8.3% vs. 89 ± 7.6% in control (P < 0.05) in A. baeri and 23 ± 11% vs 53 ± 8.3% in control (P < 0.05) in A. ruthenus. The motility pattern of the surviving frozen/thawed spermatozoa was the same as in fresh spermatozoa.     

Vasopressin-enhanced urea transport by rat inner medullary collecting duct cells in culture

Abstract. The distal inner medullary collecting duct (IMCD) is critical in the urinary concentrating process, in part because it is the site of vasopressin (AVP)-regulated permeability to urea. The purpose of these experiments was to develop a cell culture model of the IMCD on permeable structure and to characterize the responsiveness to AVP. Rat IMCD cells were grown to confluence on collagen-coated Millipore filters glued onto plastic rings. To assess the time required to achieve confluence, the transepithelial resistance was measured periodically and was found to be stable after 2 weeks, at a maximal value of 595 ± 22 ω cm². In separate monolayers the effect of AVP on inulin and urea permeability was determined. While inulin permeability was unchanged after AVP, urea permeability increased from 6.0 ± 0–4 to peak values of 16.0 ± 3–8(10nM),23.1 ± 3–9(1 μM)and28 1 ± 4–9(10μM) X 10^-6cms^-1 ( n = 24). In 10 other monolayers, after the addition of 1 mM 8-Br-cAMP, urea permeability increased from 5.1 ±0–3 to 8.1 ± 1–6 times 10^-6 cm s^-1 and, after 8-Br-cAMP +3-isobutyl-l-methylxanthine, to 12.2 ± 0–7 times 10^-6 cms^-1. We conclude that rat IMCD cells grown in culture exhibit the characteristics of a 'tight' epithelium. Inulin and urea permeability are not different in the absence of AVP, consistent with high resistance junctional complexes. Furthermore, IMCD cells retain the capacity for AVP-regulated urea permeability, a characteristic feature of this nephron segment in vivo.     

三疣梭子蟹蜕皮周期中MIH基因mRNA水平与蜕皮激素浓度变化

甲壳动物的蜕皮过程被认为是由位于眼柄的X器-窦腺复合体(XO-SG)分泌蜕皮抑制激素(MIH)通过调节Y器(YO)合成蜕皮激素而调控的。通过实时荧光定量PCR(qRT-PCR)发现MIH基因在三疣梭子蟹眼柄X器-窦腺复合体中表达最强。采用qRT-PCR分析了MIH基因在三疣梭子蟹蜕皮周期中的表达变化, 结果表明; A期为(0.42±0.08)倍, B期为(1.09±0.09)倍, C期为(1.35±0.16)倍, D₀亚期为(1.00±0.10)倍, D₁亚期(0.78±0.07)倍, D₂亚期为(0.27±0.08)倍, D_3/4亚期为(0.20±0.04)倍。采用高效液相色谱-电喷雾串联质谱(LC-MS/MS)法完成了三疣梭子蟹蜕皮周期中蜕皮激素(20E)浓度变化的测定。A/B期蜕皮激素的浓度较低, 低于仪器检测限0.33 pg, C期为(1.666±0.762) ng/mL, D₀亚期为(4.047±1.5133) ng/mL, D₁亚期为(6.756±4.928) ng/mL, D₂亚期为(8.609±3.827) ng/mL, D₃亚期为(19.534±4.799) ng/mL, D₄亚期为11.616 ng/mL。在三疣梭子蟹蜕皮周期中, MIH基因表达量与血淋巴中蜕皮激素浓度呈现一定拮抗性, 揭示MIH抑制Y器合成蜕皮激素而调控着三疣梭子蟹蜕皮的发生和进行。     

Monoamine oxidase activity in kidney and heart of Piaractus mesopotamicus (Holmberg)

The values of Michaelis–Menten constant (K_M) and maximum velocity (V_MAX) for kidney and heart monoamine oxidase (MAO) from pacu Piaractus mesopotamicus were determined. The mean ± s . e . K_M values were 17·28 ± 2·27 μM for kidney and 15·38 ± 1·86 μM for heart. MAO activities were 111·60 ± 3·25 and 15·12 ± 0·30 nmols min⁻¹ g⁻¹ of wet tissue for kidney and heart, respectively. In addition, MAO inhibitory studies in these two tissues indicate that this enzyme may be a different isoform of MAO.