健脾消癌方对结肠癌HCT116/5-Fu耐药细胞增殖、凋亡、周期的影响研究

目的：研究健脾消癌方对HCT116/5-Fu耐药细胞增殖、凋亡、周期的影响。方法：采用5-Fu浓度递增间断刺激法诱导建立HCT116/5-Fu细胞,制备健脾消癌方含药血清,设立10%、15%、20%含药血清组,设立相同浓度无药血清对照组及10%胎牛血清组。采用CellTiter GLO法检测细胞增殖能力并计算细胞抑制率,采用PI-FACS法检测细胞周期变化,采用Annexin V-APC单染法检测细胞凋亡水平。结果：与对照组相比,不同浓度含药血清组对HCT116/5-Fu细胞增殖均有抑制作用,差异均有统计学意义（P＜0.05或P＜0.01）,且随含药血清浓度的升高而抑制作用增强;与对照组相比,不同浓度含药血清组HCT116/5-Fu细胞凋亡率均明显提高,差异有统计学意义（P＜0.01）,且随含药血清浓度升高细胞凋亡率提高幅度增大;与对照组相比,不同浓度含药血清组均可将细胞周期阻滞在G0/G1期,差异有统计学意义（P＜0.01）。结论：健脾消癌方可抑制HCT116/5-Fu细胞的增殖,促进HCT116/5-Fu细胞的凋亡,阻滞其细胞周期进程,且其作用随浓度的升高而明显增强,具有改善结直肠癌耐药的作用。     

疏肝健脾解毒方含药血清对MDA-MB-231乳腺癌细胞凋亡及Bax、Bcl-2的影响

目的：研究疏肝健脾解毒方含药血清对MDA-MB-231三阴性乳腺癌（TNBC）细胞凋亡及相关X蛋白（Bax）、B淋巴细胞瘤-2（Bcl-2）蛋白表达的影响。方法：给大鼠灌胃疏肝健脾解毒方制备含药血清,设置空白对照组、含药血清低剂量组、含药血清中剂量组、含药血清高剂量组及阳性对照组,以流式细胞术检测细胞凋亡,Western Blot测定Bcl-12、Bax蛋白表达量。结果：各给药组凋亡率及Bax、Bcl-12蛋白表达水平与空白对照组比较,差异均有统计学意义（P<0.05）;含药血清低、中、高剂量组Bax、Bcl-2蛋白表达水平与阳性对照组比较,差异均有统计学意义（P<0.05）。结论：疏肝健脾解毒方对MDA-MB-231 TNBC细胞具有凋亡诱导作用,其机制可能与干预Bax、Bcl-2蛋白表达有关。     

降气平喘方对哮喘大鼠气道平滑肌细胞中ADAM33和ETS-1表达的影响

目的：探讨降气平喘方含药血清对哮喘大鼠气道平滑肌细胞(ASMC)中易感基因解整合素金属蛋白酶 33（ADAM33）和E26 转录因子-1（ETS-1）表达的影响及其作用机制。方法：通过卵清蛋白(OVA)致敏建立大鼠哮喘模型,另取SD雌性大鼠分组（降气平喘方组、孟鲁司特钠组）灌胃给药制备不同浓度的含药血清,即降气平喘方组血清分别配制为浓度5%、10%、15%的中药含药血清培养液,孟鲁司特钠组血清配制为30%浓度的西药含药血清培养液,分别对体外培养的ASMC进行同化干预24h。采用CCK8法测定不同含药血清对ASMC生长的抑制率,ELISA法检测各组细胞培养上清液中转化生长因子-β1(TGF-β1)、白细胞介素-6(IL-6)的浓度,Western Blot法检测各组细胞中ADAM33、ETS-1蛋白表达情况。结果：培养出的细胞在细胞免疫荧光检测下鉴定成功;5%和10%中药血清组均能明显抑制ASMC的增殖(P<0.05或P<0.01);ASMC培养上清液中,10%中药血清组的TGF-β1和IL-6浓度较哮喘模型组明显降低(P<0.05);与哮喘模型组比较,10%中药血清组对ADAM33蛋白的高表达下调作用更显著（P<0.01）,10%、15%中药血清组及30%西药血清组对ETS-1蛋白的高表达下调更显著（P<0.01）。结论：降气平喘方可通过调控ADAM33和转录因子ETS-1表达,减少炎性细胞介质TGF-β1和IL-6释放以及改善气道重塑,达到干预和治疗哮喘的作用。     

基于网络药理学及实验验证探究金水六君煎对人支气管上皮细胞黏液高分泌的影响

目的：通过网络药理学挖掘金水六君煎治疗气道黏液高分泌的潜在靶点,并采用体外实验探究金水六君煎对人支气管上皮细胞黏液高分泌的干预作用及作用机制。方法：采用网络药理学预测金水六君煎治疗气道黏液高分泌潜在靶点。将24只雄性SPF大鼠随机分为2组,中药组与正常组分别用金水六君煎［5.26g/（kg·d）］和等体积0.9%氯化钠注射液灌胃,5 d后麻醉下取大鼠含药血清和正常血清;将第5代人支气管上皮细胞分为正常组、模型组、不同浓度（10%、15%、20%）含药血清组、阿奇霉素组;正常组予常规培养液培养12 h后,加入15%正常大鼠血清干预12 h;其余各组采用香烟提取物（CSE）联合脂多糖（LPS）干预12h构建人支气管上皮细胞黏液高分泌模型,模型组干预后,加入15%正常大鼠血清干预12 h;不同浓度含药血清组干预后,分别加入10%、15%、20%含药血清继续干预12 h;阿奇霉素组干预后,加入含25μmol/L阿奇霉素血清继续干预12 h。采用CCK8检测细胞活力;采用Western blot法验证潜在靶点。结果：网络药理学预测黄芩素、松柏苷是与疾病靶点连接最广泛的活性物质,蛋白质-蛋白质相互作用（PPI）网络与分子对接发现关键靶点可能为基质金属蛋白酶9（MMP9）、血管内皮生长因子（VEGF）。实验研究结果显示,干预后各组细胞活性较正常组降低（P<0.05）,15%含药血清组和阿奇霉素组细胞活性较模型组升高（P<0.05）,10%、20%含药血清组细胞活性较阿奇霉素组低（P<0.05）。黏蛋白5AC（MUC5AC）是黏液高分泌的重要蛋白,其表达增加代表模型建立成功。干预后各组MMP9、VEGF、MUC5AC蛋白相对表达量较正常组增加（P<0.05）,15%含药血清组和阿奇霉素组MMP9、VEGF、MUC5AC蛋白相对表达量较模型组降低（P<0.05）。结论：金水六君煎对MUC5AC、MMP9、VEGF呈现抑制作用,揭示其机制可能是通过多靶点、多成分作用于气道黏液高分泌。     

健脾消癌方对人结肠癌HCT116细胞AKT、mTOR表达的影响

目的：探讨健脾消癌方（JPXA）含药血清对人结肠癌HCT116细胞丝氨酸/苏氨酸蛋白激酶（AKT）、哺乳动物雷帕霉素靶蛋白（mTOR）表达以及对细胞增殖、细胞周期及凋亡的影响。方法：取不同浓度的JPXA含药血清作用于HCT116细胞,筛选JPXA含药血清处理HCT116细胞的半抑制浓度（IC50）,并将HCT116细胞分为实验组和对照组,予以相应处理后,采用CCK-8法检测细胞增殖能力,PI-FACS法检测细胞周期变化,Annexin V-APC 单染法流式细胞仪检测凋亡水平,并采用Rt-PCR检测AKT mRNA、mTOR mRNA水平,采用Western Blot检测AKT、p-AKT、mTOR及磷酸化哺乳动物雷帕霉素靶蛋白（p-mTOR）水平。结果：与对照组相比,实验组HCT116细胞增殖能力显著下降,能使细胞周期停留在G1期及S期,且细胞凋亡水平上升,差异均有统计学意义（P<0.05）。与对照组相比,实验组AKT mRNA、mTOR mRNA表达水平及AKT、p-AKT、mTOR、p-mTOR蛋白含量明显下降（P<0.05或P<0.01）。结论：JPXA能抑制AKT、mTOR表达,对结肠癌细胞的增殖、克隆能力有一定抑制作用,并可在一定程度上促进结肠癌细胞的凋亡。     

加味大黄䗪虫丸对原发性胆汁性胆管炎肝星状细胞损伤状态下TGF-β1、Smads的影响

目的：观察加味大黄?虫丸对原发性胆汁性胆管炎（PBC）肝星状细胞损伤状态下转化生长因子-β1（TGF-β1）、Smads的影响。方法：将20只大鼠随机分为干预组与对照组,每组各10只。干预组灌胃加味大黄?虫丸混悬液1.5 ml/100 g,对照组灌胃等量0.9%氯化钠注射液,每天1次,持续1周。将肝星状细胞T6（HSC-T6）细胞分为6组,分别为空白组、对照组、5%空白血清组、10%空白血清组、5%含药血清组、10%含药血清组,培养24 h。建立乳酸损伤模型,造模成功后分为5组：空白组、对照组、正常组、模型组、中药组。测定细胞活性,细胞中TGF-β1、Smad3、Smad7 mRNA表达,TGF-β1、磷酸化Smad3、磷酸化Smad7 mRNA蛋白表达。结果：各组HSC-T6细胞活性对照组、5%空白血清组、10%空白血清组、5%含药血清组、10%含药血清组与空白组比较,差异均有统计学意义（P<0.01）。TGF-β1、Smad3、Smad7 mRNA表达及TGF-β1、p-Smad3、p-Smad7 mRNA蛋白表达水平,模型组与空白组比较,中药组与模型组比较,差异均有统计学意义（P<0.01或P<0.05）。结论：加味大黄土?丸治疗PBC的作用机制与降低TGF-β1、Smad3水平,提高Smad7水平相关。     

马钱子碱通过调控PI3K/Akt/mTOR通路对肺腺癌细胞的抑制作用研究

目的：探究马钱子碱对非小细胞肺癌的影响及其作用机制。方法：采用CCK-8法和划痕试验法研究马钱子碱抑制A549细胞的增殖、迁移能力,Western blot法检测各组细胞磷脂酰肌醇3激酶/蛋白激酶B/哺乳动物雷帕霉素靶蛋白(PI3K/Akt/mTOR)通路相关的蛋白表达,ELISA法检测白细胞介素6（IL-6）、肿瘤坏死因子-α（TNF-α）的含量。结果：CCK-8试验结果显示,给药组对A549细胞增殖有明显抑制作用;划痕试验显示,与空白组比较,给药组细胞迁移率明显下降（P<0.01）;Western blot结果显示,给药干预后可显著下调p-Akt、p-mTOR的表达（P<0.001）,同时给药组的mTOR、Akt、PI3K蛋白表达下调（P<0.05）;ELISA试验结果显示,采用马钱子碱进行干预后,细胞上清IL-6和TNF-α含量均显著升高(P<0.01或P<0.001)。结论：马钱子碱可有效抑制非小细胞肺癌细胞的增殖、迁移,可能是通过抑制PI3K/Akt/mTOR通路而起作用,并通过调节TNF-α含量来达到抗肿瘤的作用。     

肝宁方对NASH小鼠保护作用及大鼠肝星状细胞影响的实验研究

目的：探讨肝宁方对非酒精性脂肪性肝炎（NASH）小鼠的保护作用及其对活化后大鼠肝星状细胞（HSCs）的影响。方法：将24只小鼠随机分为正常组、模型组、肝宁方组,每组各8只。对模型组、肝宁方组采用高脂饮食联合四氯化碳豆油溶液皮下注射建立NASH小鼠模型,造模成功后正常组和模型组予0.9%氯化钠注射液灌胃,肝宁方组予肝宁方灌胃。干预3周。将大鼠HSCs分成正常组、模型组及肝宁方组,正常组予正常培养基培养,模型组及肝宁方组采用转化生长因子-β1（TGF-β1）活化,随后正常组和模型组用含正常大鼠血清培养基培养,肝宁方组加入含30%肝宁方含药血清培养基培养。培养24 h。观察肝组织病理学情况;血清天冬氨酸氨基转移酶(AST)、丙氨酸氨基转移酶(ALT)、血清总胆固醇（CHOL）、三酰甘油（TG）水平;肝组织中肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6）表达,葡萄糖调节蛋白78（GRP78）及内质网核信号转导蛋白1α（IRE-1α）蛋白表达;肝细胞凋亡情况;HSCs的增殖情况。结果：肝宁方组和模型组小鼠相比,脂质空泡和出血减少,炎性细胞浸润减轻。小鼠血清AST、ALT、CHOL、TG水平,肝组织TNF-α、IL-6水平,GRP78、IRE-1α蛋白表达,肝细胞凋亡情况模型组与正常组比较,肝宁方组与模型组比较,差异均有统计学意义（P<0.01或P<0.05）。与模型组相比,肝宁方含药血清干预后HSCs增殖趋势明显减弱（P<0.01）。结论：肝宁方能够改善NASH小鼠肝脏病理学变化,保护肝脏、调节脂质代谢、抑制炎症反应、减轻肝细胞凋亡、抑制HSCs增殖,其机制可能与调控内质网应激有关。     

鹿龙再生汤对肺癌裸鼠化疗骨髓抑制期骨髓的保护作用

目的：探讨鹿龙再生汤对肺癌裸鼠化疗骨髓抑制期骨髓的保护作用。方法：采用人肺鳞癌细胞株Eca109进行肺癌肿瘤造模,再腹腔注射环磷酰胺（CTX）进行肿瘤化疗骨髓移植造模。造模成功后分为4组。空白对照组10只,模型组、西药组、中药组各15只。西药组予利可君片灌胃,重组人粒细胞刺激因子一次性皮下注射;中药组予鹿龙再生汤灌胃,空白对照组和模型组不予药物干预。观察各组的白细胞（WBC）、血小板（PLT）、红细胞（RBC）、血红蛋白（HB）水平和骨髓有核细胞数（BMNC）;骨髓活检观察骨髓增殖情况;流式细胞仪检测细胞凋亡,免疫组织化学技术检测细胞凋亡因子FAS抗体表达水平。结果：模型组、西药组、中药组WBC、PLT、RBC、HB水平及BMNC与空白对照组比较,差异均有统计学意义（P＜0.05）;西药组、中药组给药3、5 d WBC、PLT、RBC水平及BMNC与模型组比较,差异均有统计学意义（P＜0.05）。中药组较模型组显示出较好的骨髓增殖效果;中药组骨髓单个核细胞生存比例及FAS抗体表达水平较模型组升高。结论：鹿龙再生汤对肺癌裸鼠化疗骨髓抑制期骨髓粒细胞系统、巨噬细胞系统、红细胞系统抑制均有促进细胞增殖的作用,其作用机制可能与调控细胞凋亡因子FAS抗体表达有关。     

月华丸（胶囊）及其拆方对巨噬细胞增殖影响的研究

目的：研究月华丸（胶囊）对小鼠单核巨噬细胞（RAW264.7）活性的影响,为后续研究月华丸（胶囊）对耐多药结核分支杆菌感染细胞自噬的影响提供前期实验基础。方法：取对数生长期的RAW264.7细胞,用10%胎牛血清培养制成的细胞悬液加入96孔培养板中培养。分别采用终浓度为1.25g/mL、0.63g/mL、0.32g/mL、0.16g/mL、0.125g/mL、0.063g/mL、0.032g/mL、0.016g/mL不含阿胶的月华丸（胶囊）以及相同浓度梯度含阿胶的月华丸（胶囊）处理RAW264.7细胞,并设不加药的对照组。以4-甲偶氮唑蓝（MTT）法检测细胞存活情况。结果：（1）不含阿胶的月华丸（胶囊）在1.25g/mL、0.63g/mL浓度时对RAW264.7细胞增殖抑制率明显升高,与对照组比较差异有统计学意义（P<0.05）,表明不含阿胶的月华丸（胶囊）高浓度对巨噬细胞的增殖有一定的抑制作用。（2）含阿胶的月华丸（胶囊）对RAW264.7细胞增殖率明显升高,各浓度组与对照组比较差异均有统计学意义（P<0.05）,提示含阿胶的月华丸（胶囊）对巨噬细胞的增殖有明显促进作用。（3）浓度为0.32g/mL的含阿胶月华丸（胶囊）对RAW264.7细胞增殖率最高,为最佳促细胞增长药液浓度。结论：月华丸（胶囊）能提高巨噬细胞的活力,增强巨噬细胞的增殖。     

Model of the post-Cenozoic evolution of the cryolithozone of the shelf of the western part of the Laptev Sea

The analysis of the structure of the cryolithozone, facies, and thicknesses of the Quaternary sediments and the results of the physicochemical mathematical modeling of the modern shelf of the western part of the Laptev Sea support the influence of the Late Pleistocene glaciations on the heat conditions and the distribution of the permafrost in the area. A ～200-m thick glacier formed under aerial conditions from atmospheric precipitation represented the metamorphosed snow cover. According to the modeling, the long-living (from 60?50 to 10?4 ky) glacier reduced the thickness of the permafrost rocks in the reviewed shelf area for 280–360 m. The Holocene marine transgression additionally decreased the thickness from 50–140 m on the inner shelf to 220–350 m on the outer shelf. The modern submarine cryolithozone 450-0 m thick is wide-spread in the studied region from the coast to the shelf boundary (isobaths of 130–140 m), where it pinches out at a distance of ～380 km from the coast at a depth of ～250 m above the sea level.     

Investigation of the mechanism of the long-term evolution of the carbon cycle in the ecosystem of the Sevastopol bay

We analyze the components of the carbon system of the Sevastopol bay waters and the balance of main sediment-forming substances using the data of field investigations in 1998–2008. The interannual variations of total inorganic carbon and the equilibrium partial pressure of carbon dioxide in the bay water are noted. An increase in the flux of carbon dioxide into the bay and in the content of organic carbon in bottom sediments is revealed, and an explanation of this phenomenon is given. The priority accumulation of organic carbon in the sediments of the bay is established. We assess the interannual variation in the relative abundances of organic and inorganic carbon as an index of the carbon cycle stability.     

Contemporary state of the investigation of the influence of the discharge of rivers on the hydrologic structure of the Black Sea

We generalize and analyze the state of investigations of the influence of river waters on the hydrologic structure of the Black Sea. Specific features of the water, regime and hydrography of rivers flowing into the sea, data on the discharge of rivers, and intrayear and interyear variability of the discharge are presented. We discuss and analyze works studying the transformation of river waters, routes of desalinized sea waters, and their influence on the thermohaline structure and dynamic regime of the sea. Translated by Peter V. Malyshev and Dmitry V. Malyshev     

Modelling of the ecosystem of the Black-Sea shelf zone near the estuary of the Danube

We analyze the results of numerical calculations performed according to the three-dimensional interdisciplinary model of an ecological system of the Black-Sea shelf zone near the estuary of the Danube. The complete system of equations of hydrothermodynamics is solved together with transport equations of the advection-diffusion-reaction type used to describe the transformation of a substance (nitrogen) between the components of the characteristic vectors of the ecosystem: plankton, detritus, and biogenic elements (nitrates). We describe the distinctive features of the circumcontinental distribution of components obtained as a result of numerical experiments and present arguments for the conclusion that the ecosystem of the Danube estuary water area plays the role of a buffer zone between the press of the Danubian biogenic pollutions and the neighbouring areas of the shelf zone and open sea. Translated by Peter V. Malyshev and Dmitry V. Malyshev     

Dynamics of the ice cover and peculiarities of the ice transportation of the sediments at the tidal flats of the Kandalaksha Gulf of the White Sea

Ten years of semistationary studies of the interaction of the ice cover and the coastal relief of the Kandalakhsa Gulf in the White Sea allowed us to reveal the peculiarities of these processes in the warm and cold years and the mechanisms of the origination of ice forms, including ice tents, which govern the character of the tidal flats. Three dynamic fast ice zones are distinguished by their geomorphological location, morphology, and ice dynamics. The dependence of the mechanisms and volumes of the enrichment of the fast ice in the sediments on the coastal zone??s relief was established. At the coast of the Kindo Peninsula (Velikaya Salma Strait), the ice mostly contains fine-sandy sediments. Coarser sediments, including boulders, are occasional and frozen into the ice at the contact of the fast ice and the bottom. At the expense of the ice??s spreading by 4?C6 times in the warm years and 2?C4 times in the cold years, several thousand tons of sediments mostly from the tidal flat??s surface (100?C700 m³ from 1 km of coast in the wide tidal flats and <50 m³ on the steep coasts) are delivered to the water area.     

Relationship between the path of the Kuroshio in the south of Japan and the path of the Kuroshio Extension in the east

A relationship between paths of the Kuroshio and Kuroshio Extension (KE) is investigated, using the satellite-derived altimetry dataset of 1993–2008. When the Kuroshio takes the nearshore nonlarge meander path or typical large meander path and resultantly goes through the deeper channel (about 2500 m) of the Izu-Ogasawara Ridge, the KE path adopts a relatively stable state with the two quasi-stationary meanders. On the other hand, when the Kuroshio takes the offshore nonlarge meander path and then passes over the shallower part of the Ridge (about 1000 m), the KE path tends to be convoluted, i.e., an unstable state.     

Variability of the nutrient runoff of the rivers of the Arctic Basin

The spatiotemporal variability of the hydrochemical composition of the riverine runoff in nearmouth areas is presented using the example of selected rivers of the Arctic Basin. The temporal dynamics of the riverine nutrient runoff are mainly determined by the seasonal and interseasonal variations in the drainage basins. The spatial variations in the chemical composition of riverine waters in the near-mouth areas are mainly caused by the dynamics of the water flow and the river bed relief and, to a lesser degree, by biochemical processes. It is shown that the variations in the nutrient composition of the waters in near-mouth areas are comparable in magnitude to those at the river-sea geochemical barriers.     

Parameters of the sediments of the deep-water basin of the Black Sea

The methodology and the first results of the computations of the volumes, masses, and growth rates of the sedimentary body of the buried Black Sea basin are presented. Their temporal evolution reflects the regional paleogeodynamics, in particular, the reorientation of the vector of the relative movement of the Arabian and Eurasian lithospheric plates and the related intensification of their collision in the late Miocene-Pliocene.     

Modelling of the ecosystem on the shelf of the Republic of Guinea

Three mathematical models are considered: a two-dimensional model for calculating the concentration fields of the ecosystem components on the shelf of the Republic of Guinea, a pointwise model for the annual variation of the components, and a two predators-prey model. These models are used to study the effect of hydrodynamic conditions on the state of the shelf ecosystem and for research into the causes which distort the balance between marketable and unmarketable fish.Translated by Mikhail M. Trufanov.     

Evolution of the slope of the buried depression of the Black Sea

The first data on the temporal evolution of the slope of the buried depression of the Black Sea are obtained. The sharp change in the rate of the decrease in the slope steepness that occurred in the Late Miocene-Pliocene time reflects a reorientation of the vector of mutual movement and a related intensification of the collision between the Arabian and Eurasian lithospheric plates.