以结果验证为导向的中医学八年制Western Blot实践教学

“实验中医学”是海军军区大学中医学八年制学员的主干课程。其中Western Blot是研究蛋白水平的基本方法,也是学员所必须掌握的实验研究方法。本课程以结果验证为导向的教学方式革新,最大限度压缩每个步骤的操作时间,聚焦于让学员完成最多的实验步骤,力求实验的完整性。该方法可在一堂课完成实验,展示结果并进行分析讨论。该教学方式有助于在最短时间内掌握完整的Western Blot实验技术,值得进一步推广应用。

Geomorphic signatures and active tectonics in western Saurashtra,Gujarat,India

Active tectonics in an area includes ongoing or recent geologic events.This paper investigates the tectonic influence on the subsidence,uplift and tilt of western Saurashtra through morphotectonic analysis of ten watersheds along with characteristics of relief and drainage orientation.Watersheds 7-9 to the north(N)are tectonically active,which can be linked with the North Kathiawar Fault System(NKFS)and followed by watersheds 6,10,1,4 and 5.Stream-length gradient index and sinuosity index indicate the effect of tectonic events along the master streams in watersheds 6-9.Higher R^(2)values of the linear curve fit for watershed 7 indicate its master stream is much more tectonically active than the others.The R^(2)curve fitting model and earthquake magnitude/depth analysis confirm the region to be active.The reactivation of the NKFS most likely led to the vertical movement of western Saurashtra.

Food Hygiene and Safety Practices amongst Food Vendors in the Western Area, Sierra Leone

Food safety and hygiene practices require a multisectoral approach including food, water, sanitation, waste management, transport, education, trade, policies and programs that enable safe food preparation, storage and service. Unsafe food can cause illness keeping people from achieving their full potential and death. This was a descriptive study that uses a mixed method approach to derive insights into the characteristics of food vendors related to demography, knowledge, practices, infrastructure, compliance and recommendation for a policymaking framework. Using the Lemeshows’ sample size formula, 473 vendors from formal (restaurants) and informal (cookri-baffa/table top) sites were interviewed and observed. We found from discussions that respondents had a good understanding on how to keep food safe. However, observed practices were poor 93% handled food with their bare hands, 83% did not cover their hair, and 76% did not wear an apron whilst handling, preparing or serving food, 61% did not keep their finger nails clean or short and 57% did not wash their hand before preparing or serving food. Over half (51%) had access to a toilet but 32% reported their use required payment and emphasized their poor condition/inadequate management. Nearly half (47%) of the vending sites did not have a handwashing facility, with soap and water available. Only 7% reported having any authority oversight of food safety. Food safety and hygiene practices in most cookri shops and restaurants was ‘poor’ with very limited surveillance system in place by competent authorities for compliance of food operators. Hand washing, clean surroundings, and covered food were the most common and emphasized practices to mitigate the risks associated with unsafe food.

Impacts of hydropower-induced flow alterations on composition and diversity of riparian vegetation in the Western Himalayas: A case study in Uttarakhand, India

The increasing demand for water and energy resources has led to widespread dam construction,particularly in ecologically sensitive regions like the Himalayan Range.This study focuses on the Uttarakhand state in the Western Himalayas,where hydroelectric projects(HEPs)have significantly altered river flow regimes.The research investigates the impact of flow alterations on the composition and structure of riparian vegetation in the Garhwal Himalayas,specifically analysing four rivers regulated by hydroelectric projects.Utilizing the paired-reach comparison method,control(undisturbed),diverted(downstream of barrage/dam),and altered flow conditions(downstream of water outlet)were examined.The research reveals diverse and unique riparian ecosystems,with 89 genera and 113 taxa identified,showcasing the dominance of families like Asteraceae and Lamiaceae.The study unveils the structural importance of key species such as Berberis asiatica and Artemisia nilagirica.The density,diversity,and richness of shrub and herb species vary significantly across flow conditions.Notably,altered flow conditions demonstrate resilience in vegetation structure,while diverted conditions exhibit decreased species richness and density.The study emphasizes the importance of nuanced environmental flow management for mitigating adverse effects on riparian biodiversity in the fragile Himalayan region.These findings contribute to the global discourse on dam impacts and riparian ecology,shedding light on the complexities of this dynamic relationship in a vulnerable ecosystem.

Track-Pattern-Based Characteristics of Extratropical Transitioning Tropical Cyclones in the Western North Pacific

Based on the Regional Specialized Meteorological Center(RSMC)Tokyo-Typhoon Center best-track data and the NCEP-NCAR reanalysis dataset,extratropical transitioning(ET)tropical cyclones(ETCs)over the western North Pacific(WNP)during 1951–2021 are classified into six clusters using the fuzzy c-means clustering method(FCM)according to their track patterns.The characteristics of the six hard-clustered ETCs with the highest membership coefficient are shown.Most tropical cyclones(TCs)that were assigned to clusters C2,C5,and C6 made landfall over eastern Asian countries,which severely threatened these regions.Among landfalling TCs,93.2%completed their ET after landfall,whereas 39.8%of ETCs completed their transition within one day.The frequency of ETCs over the WNP has decreased in the past four decades,wherein cluster C5 demonstrated a significant decrease on both interannual and interdecadal timescales with the expansion and intensification of the western Pacific subtropical high(WPSH).This large-scale circulation pattern is favorable for C2 and causes it to become the dominant track pattern,owning to it containing the largest number of intensifying ETCs among the six clusters,a number that has increased insignificantly over the past four decades.The surface roughness variation and three-dimensional background circulation led to C5 containing the maximum number of landfalling TCs and a minimum number of intensifying ETCs.Our results will facilitate a better understanding of the spatiotemporal distributions of ET events and associated environment background fields,which will benefit the effective monitoring of these events over the WNP.

Quantitative effect of kerogen type on the hydrocarbon generation potential of Paleogene lacustrine source rocks,Liaohe Western Depression,China

Kerogen types exert a decisive effect on the onset and capacity of hydrocarbon generation of source rocks.Lacustrine source rocks in the Liaohe Western Depression are characterized by thick deposition,high total organic carbon(TOC)content,various kerogen types,and a wide range of thermal maturity.Consequently,their hydrocarbon generation potential and resource estimation can be misinterpreted.In this study,geochemical tests,numerical analysis,hydrocarbon generation kinetics,and basin modeling were integrated to investigate the differential effects of kerogen types on the hydrocarbon generation potential of lacustrine source rocks.Optimized hydrocarbon generation and expulsion(HGE)models of different kerogen types were established quantitatively upon abundant Rock-Eval/TOC/vitrinite reflectance(R_(o))datasets.Three sets of good-excellent source rocks deposited in the fourth(Es4),third(Es3),and first(Es1)members of Paleogene Shahejie Formation,are predominantly types I-II_(1),II_(1)-II_(2),and II-III,respectively.The activation energy of types I-II_(2)kerogen is concentrated(180-230 kcal/mol),whereas that of type III kerogen is widely distributed(150-280 kcal/mol).The original hydrocarbon generation potentials of types I,II_(1),II_(2),and III kerogens are 790,510,270,and 85 mg/g TOC,respectively.The Ro values of the hydrocarbon generation threshold for type I-III source rocks gradually increase from 0.42%to 0.74%,and Ro values of the hydrocarbon expulsion threshold increase from 0.49%to 0.87%.Types I and II_(1)source rocks are characterized by earlier hydrocarbon generation,more rapid hydrocarbon expulsion,and narrower hydrocarbon generation windows than types II_(2)and III source rocks.The kerogen types also affect the HGE history and resource potential.Three types(conventional,tight,and shale oil/gas)and three levels(realistic,expected,and prospective)of hydrocarbon resources of different members in the Liaohe Western Depression are evaluated.Findings suggest that the Es3 member has considerable conventional and unconventional hydrocarbon resources.This study can quantitatively characterize the hydrocarbon generation potential of source rocks with different kerogen types,and facilitate a quick and accurate assessment of hydrocarbon resources,providing strategies for future oil and gas exploration.

Introduction of Western Medicines into China and Improvement on Decoctions during Modern China

During the late Qing dynasty(1840 A.D.-1912 A.D.),a large quantity of Western medicines entered China,which continuously impacted the traditional Chinese medicine(TCM)market and revealed the shortcomings of Chinese medicines.Some personages in the TCM community followed the trend of learning from the West,and attempted to reform TCM,with the improvement on decoction becoming an important aspect of this effort.Through debates and trials,the improvement on decoction underwent three stages of conceptual evolution:“taking Chinese medicines as the foundation and referring to the dosage forms of Western medicines”,“introducing Western techniques to serve the preparation of decoctions”and“integrating the theories of TCM and Western medicine to improve decoctions”.The study highlights the effective complementarity between modern TCM and Western medicine in the field of pharmacy,and provides valuable experience and support for the reevaluation of the value of TCM in contemporary society.

Morphological and molecular description of a new species of sandfly,Sergentomyia(Neophlebotomus)ashwanii sp.nov.(Diptera:Psychodidae)from Western Ghats,India

Objective:To report a new species of sandfly,Sergentomyia(Neophlebotomus)ashwanii sp.nov.(Diptera:Psychodidae)from Western Ghats,India.Methods:A systematic sandfly survey was conducted in the Thrissur and Kollam districts of Kerala,India using mechanical aspirators,light and sticky traps,both indoor and outdoor habitats,for a period of one year.Deoxyribonucleic acid barcoding of samples was performed targeting mitochondrial cytochrome oxidase I(COI)gene and sequence generated was subjected to phylogenetic analysis.Results:Sergentomyia(Neophlebotomus)ashwanii,a new sandfly species is recorded and described in this communication.A single row of 10-12 pointed teeth in the cibarium with 4-6 small denticles or fore-teeth are the key characteristics that is distinctive from other members of the subgenus Neophlebotomus.Mitochondrial COI barcode followed by phylogenetic analysis of the nucleotide sequence confirms that specimens of the species belong to the same taxonomic group while the genetic distance(14.2%)with the congeners established it to be a different species.Conclusions:The Western Ghats'being an important biodiversity hotspot and has dearth of systematic entomological surveys on sandflies.The current study tried to fill the void and also report a new sandfly species.

Genesis and tectonic setting of Early Jurassic granitic rocks in Huashan Town,Xingcheng,western Liaoning

There are a large number of Mesozoic intrusive and volcanic rocks in western Liaoning of China,which is an ideal place to study the Mesozoic Paleo-Pacific subduction processes,and lithospheric destruc-tion of North China Craton.Detailed petrographic,zircon U-Pb dating and geochemical studies of the Early Jurassic granites in Huashan pluton,Xingcheng,western Liaoning,indicate that the Early Jurassic granites were formed at 184-174 Ma,mainly composed of syenite and monzogranite.The geochemical characteristics show high contents of SiO_(2),Al_(2)O_(3)and Na_(2)O+K_(2)O,low contents of Fe_(2)O_(3)and MgO,enrichment in LREEs and LILEs,and depletion in HREEs and HFSEs,and have a high content of Sr and low contents of Y and Yb,with weak negative Eu anomalies and slightly negative anomalies of Ce,indicating that they are a set of intermediate-acidic adakitic granites in high-K calc-alkaline series.All the facts significantly suggest that the Early Jurassic adakitic granites were formed at the active continental margin in the context of Paleo-Pacific plate subduction.

Spatiotemporal Changes of Snow Depth in Western Jilin,China from 1987 to 2018

Seasonal snow cover is a key global climate and hydrological system component drawing considerable attention due to glob-al warming conditions.However,the spatiotemporal snow cover patterns are challenging in western Jilin,China due to natural condi-tions and sparse observation.Hence,this study investigated the spatiotemporal patterns of snow cover using fine-resolution passive mi-crowave(PMW)snow depth(SD)data from 1987 to 2018,and revealed the potential influence of climate factors on SD variations.The results indicated that the interannual range of SD was between 2.90 cm and 9.60 cm during the snowy winter seasons and the annual mean SD showed a slightly increasing trend(P>0.05)at a rate of 0.009 cm/yr.In snowmelt periods,the snow cover contributed to an increase in volumetric soil water,and the change in SD was significantly affected by air temperature.The correlation between SD and air temperature was negative,while the correlation between SD and precipitation was positive during December and March.In March,the correlation coefficient exceeded 0.5 in Zhenlai,Da’an,Qianan,and Qianguo counties.However,the SD and precipitation were neg-atively correlated over western Jilin in October,and several subregions presented a negative correlation between SD and precipitation in November and April.

Research Progress on Integrated Chinese and Western Medicine Treatment of Tourette Syndrome in Children

Tourette syndrome is a common neuropsychiatric disorder that affects the physical and mental health of children.Early detection,diagnosis,and treatment are crucial to prevent serious impacts on the affected children,their families,and society.In recent years,there has been an increasing trend towards using a combination of methods in the clinical treatment of children with Tourette syndrome.This approach has achieved remarkable results,leading to a reduction in the rate of Tourette syndrome symptoms in children.In recent years,a combination of clinical methods has been used to treat children with Tourette syndrome,resulting in significant improvement in control rates.This article reviews the etiology of infantile Tourette syndrome and the progress made in Chinese and Western medicine treatments,providing a reference for further treatment of the condition.

A teleconnection pattern of upper-tropospheric circulation anomalies over the Eurasian continent associated with the interannual variability of atmospheric convection over the tropical western North Pacific in July

热带西北太平洋大气对流的年际变化对东亚夏季气候有着显著影响,而且受到前冬ENSO的影响.然而,还有相当多的西北太平洋对流年际变化不能由前冬ENSO解释.本文表明,7月西北太平洋对流受到欧亚大陆对流层上层遥相关型的显著影响,且这一影响与前冬ENSO的影响程度相当.该遥相关型表现为位于欧洲的气旋式环流异常,中亚的反气旋式环流异常和东亚带状延伸的气旋式环流异常,并表现为7月对流层上层环流的第一主模态.基于本文的结果并结合前人的研究,作者认为这一遥相关型能通过亚洲急流出口区附近的罗斯贝波破碎引起西北太平洋对流异常.

A Case Report on Treating Progressive Supranuclear Palsy with a Combination of Chinese and Western Medicine

A case of progressive supranuclear palsy in a 66-year-old woman is presented.The patient complained of“progressive stiffness of both lower limbs for five years,accompanied by choking and coughing for more than 1 year,”and was diagnosed with paralysis in traditional Chinese medicine(TCM),which was characterized by a deficiency of liver and kidney,and phlegm and blood stasis blocking the collaterals.Western medicine diagnosed it as progressive supranuclear palsy.The patient was diagnosed with“Parkinson’s disease”several times in the past and was given“polybasic hydrazide”and“amantadine hydrochloride tablets,”etc.The patient did not have significant symptomatic relief,so she sought further treatment with a combination of traditional Chinese and Western medicine.Based on the concept of deficiency and excess,this disease is categorized as a paralytic disease in Chinese medicine,and the onset of the disease is closely related to the deficiency of the body after a long period of illness,with wind,phlegm,blood stasis,and deficiency as the main pathogenetic mechanism.The treatment is based on the combination of disease and evidence,with traditional Chinese medicine and acupuncture as the main treatments,supplemented by intermediate-frequency pulsed electrical stimulation,transcranial repetitive magnetic stimulation,medicated canisters,wax therapy,traditional Chinese medicine guasha,and acupoint injections.Western medicine is based on symptomatic treatment,and the patient’s symptoms were relieved significantly after the combination of Chinese and Western medicine.

麻疯树外植体愈伤组织中毒蛋白的Western-blot鉴定

The proteins from endosperm, root, stem, leaf, leafstalk of Jatropha curcas L. and their calli were hybridized to the seed toxin protein (curcin) of Jatropha curcas L. by western blot. The result showed that the curcin was specifically expressed in the endosperm and its calli, while it was not detected in root, stem, leaf, and leafstalk of Jatropha curcas L. and their calli . This study indicated that calli induced from endosperm can be used to produce curcin.

重组N蛋白抗原检测PRRSV抗体ELISA的研究 Ⅳ.与IDEXXELISA试剂盒及Westernblot的比较

利用重组N蛋白抗原建立了检测猪繁殖与呼吸综合征病毒(PRRSV)抗体的ELISA方法并组装成试剂盒,将试制的3批试剂盒分别与IDEXX生产的试剂盒及Western-blot进行了符合率试验。试验结果表明,所研制的试剂盒与Westernblot的符合率达97.67%以上。对460份临床血清分别用自制的试剂盒和IDEXX公司生产的试剂盒进行检测,其中有37份不相符合,用Westernblot对这37份血清进行验证,有35份血清检测结果与自制的试剂盒检测结果一致。由此表明,自行研制的试剂盒,其特异性和敏感性均能满足目前临床上该疫病的流行病学分析或免疫抗体检测。

以融合蛋白为抗原的简化Westernblot法诊断囊虫病

目的：为囊虫的诊断提供简便有效的方法。方法：猪囊尾蚴ｃＤＮＡ表达文库中筛选出的β－半乳糖苷糖－猪囊虫ｃＤＮＡ编码的融合蛋白为抗原，进行简化的Ｗｅｓｔｅｒｎｂｌｏｔ分析。结果：四个克隆的融合蛋白（ｃＣ１、ｃＣ２、ｃＰ１和ｃＨ１）联合使用，检测１２４例人囊虫病血清中的抗囊尾蚴ＩｇＧ抗体，阳性率达９３．７％，三个克隆的融合蛋白（ｃＣ１、ｃＣ２、和ｃＰ１）联合使用，检测３８例猪囊虫病血清中的相应抗体，阳性率达１００％，均高于各克隆融合蛋白单独使用的阳性率，且与其他寄生虫病血清无交叉反应。结论：以融合蛋白为抗原所进行的Ｗｅｓｔｅｒｎｂｌｏｔ分析具有高度灵敏性和特异性，且简便易操作。

新孢子虫和弓形虫ELISA及western blot检测方法的建立及应用

为建立牛新孢子虫和弓形虫的免疫学检测方法,并调查新疆部分地区牛新孢子虫和弓形虫的感染情况,本研究应用纯化的新孢子虫重组蛋白SRS2(NcSRS2)和弓形虫重组蛋白SAG2(TgSAG2)作为包被抗原,分别建立新孢子虫和弓形虫的ELISA和western blot血清学检测方法,并进行特异性和重复性试验,以其检测662份疑似样品,并与商品化试剂盒检测结果比较验证。特异性和重复性试验结果表明,建立的方法特异性强、重复性良好。采用建立的两种ELISA方法对662份临床样品的检测结果表明,新孢子虫和弓形虫的抗体阳性率分别为13.44%(89/662)和5.29%(35/662);与IDEXX试剂盒和永辉试剂盒的符合率分别为94.11%和95.92%。此外,western blot检测的新孢子虫和弓形虫抗体阳性率分别为5.14%(34/662)和3.17%(21/662);与建立的ELISA检测方法的符合率分别为91.69%和97.89%。本研究为分析奶牛流产的原因提供了一定的依据。

Western blot法诊断囊虫病的应用研究

目的：探讨四种猪囊尾蚴（Ｃｙｓｔｉｃｅｒｃｕｓｃｅｌｕｌｏｓａｅ）特异性抗原ｃＣ１、ｃＣ２、ｃＰ１、ｃＨ１（分子量分别为２８ｋＤａ、１８ｋＤａ、１４ｋＤａ、３４ｋＤａ）混合应用诊断人囊虫病的价值。方法：以猪囊尾蚴ｃＤＮＡ表达文库中筛选出的β－半乳糖苷酶－猪囊虫特异性抗原ｃＤＮＡ编码的融合蛋白（ｆｕｓｉｏｎｐｒｏｔｅｉｎｓ，ＦＰ）为抗原，作简化的Ｗｅｓｔｅｒｎｂｌｏｔ（ｉｍｐｒｏｖｅｄＷｅｓｔｅｒｎｂｌｏｔ，ＩＷＢ）分析，检测１０７例囊虫病、４０例华支睾吸虫病、２４例包虫病和３４例健康人血清对融合蛋白的反应。同时应用粗制抗原（ｃｒｕｄｅａｎｔｉ－ｇｅｎ，ＣＡ）进行ＥＬＩＳＡ、ＩＨＡ对比检测。结果：在简化Ｗｅｓｔｅｒｎｂｌｏｔ检测中，ＦＰ被１０７例囊虫病患者血清中９４例（８７．９％）血清所识别，与华支睾吸虫病、包虫病患者及健康人血清无交叉反应，特异性为１００％。以粗制抗原作ＥＬＩＳＡ、ＩＨＡ检测囊虫病人血清中的ＩｇＧ抗体阳性率分别为８４．１％和７４．８％，与华支睾吸虫病患者血清存在交叉反应，假阳性率分别为２．５％、１２．５％；与包虫病患者血清存在交叉反应，假阳性率分别为８．３％、１６．７％；与健康人血清也存在交?

定量PCR和Western blot结合检测回药爱康方对肺癌细胞EGFR、NF-κB的表达影响

目的:探讨定量PCR和Western blot结合检测对Lewis肺癌C57小鼠瘤细胞表皮生长因子受体(EGFR)、NF-κB表达影响,观察该方对调节癌细胞信号转导作用机制。方法:小鼠自接种瘤株细胞24 h后称重、编号,随机分为8组,每组5只,分别为:1回药高剂量组(AKH);2荷瘤模型组(对照组);3回药中剂量组(AKM);4单纯化疗药组(CTX组);5回药中剂量组加化疗组(AKM+CTX);6回药低剂量加化疗药组(AKL+CTX);7回药低剂量组(AKL);8回药高剂量加化疗药组(AKH+CTX)。采用RT-PCR和western blot方法检测小鼠组织样本中EGFR、NF-κBm RNA转录水平和蛋白质表达的变化情况。结果:实时荧光定量PCR结果显示:与模型组小鼠比较,给予回药爱康方,高、中、低剂量干预后,小鼠瘤细胞中EGFR m RNA表达水平显著下调(P<0.01),而低剂量对NF-κBm RNA的表达水平无统计学差异(P>0.05),给予回药爱康方,高+CTX、中+CTX、低+CTX、CTX后,小鼠瘤细胞中EGFR表达水平显著下调(P<0.01),小鼠瘤细胞中NF-κBm RNA表达水平显著上调(P<0.01);与模型组比较,给予回药爱康方高、中、低剂量干预后,Lewis肺癌C57小鼠EGFR、NF-κB蛋白表达水平无统计学差异性(P>0.05),给予回药爱康方高+CTX、中+CTX、低+CTX、CTX干预后,Lewis肺癌C57小鼠EGFR蛋白表达水平显著下调、NF-κB蛋白表达水平显著上调(P<0.01或P<0.05)。结论:回药爱康方高、中、低与回药开康方高+CTX、中+CTX、低+CTX、CTX均对EGFRm RNA的表达水平有影响,仅低剂量对NF-κBm RNA的表达水平无影响,同时回药爱康方高、中、低剂量均对EGFR、NF-κB蛋白表达无影响。

快慢转法及不同滤膜和显色检测法在Western blotting中的应用分析

目的探讨快、慢转法及不同滤膜和显色检测法在Western blotting中的应用及各实验环节分析。方法采用MDA-MB-231细胞制备细胞蛋白,应用不同印迹膜[硝酸纤维素膜、聚偏乙烯二氟(PVDF)膜和阳离子尼龙膜]分别采用快转法和慢转法进行增强化学发光法(ECL)和DAB化学显色法检测,并对Western blotting的各实验环节进行了分析。结果(1)在快、慢转法中硝纤膜的蛋白预染marker条带略强于尼龙膜,而尼龙膜又略强于PVDF膜;PVDF膜和尼龙膜的正反面易于混淆,而硝纤膜的正反面不易混淆。(2)在快、慢转法中,PVDF膜的DAB化学显色法图像略强于尼龙膜和硝纤膜,而尼龙膜和硝纤膜无明显差异;与慢转法相比,快转的硝纤膜和尼龙膜中的蛋白条带略呈波浪状。(3)在慢转法中三种膜化学发光法的图像无明显背景,但在快转法中尼龙膜的背景较明显,而硝纤膜和PVDF膜亦无明显背景。结论应根据实验需要选用不同的印迹膜;慢转法通常优于快转法,增强化学发光法优于DAB化学显色法;增强化学发光法特异胜强、灵敏度高,是分析蛋白质表达较为理想的方法。