上皮特异性抗原和CD44蛋白在原发性结直肠癌组织中的表达及临床意义

目的分析免疫组织化学链霉菌抗生物素蛋白-过氧化物酶(SP)法测定原发性结直肠癌组织中上皮特异性抗原(ESA)和CD_(44)蛋白表达的关系及临床病理意义。方法选择2012年1月至2014年1月来中国医科大学附属盛京医院检查并确诊的结直肠癌患者52例(结直肠癌组),选择同期来院检查的正常结直肠黏膜组织患者20例(正常黏膜组)及伴不典型增生的结直肠腺瘤20例(不典型增生组),采用免疫组织化学SP法检测对三组ESA和CD_(44)蛋白表达水平。结果在年龄>50岁、肿瘤直径>3 cm、浸润深度至浆膜及Dukes分期C+D期的ESA阳性表达率高于年龄≤50岁、肿瘤直径≤3 cm、浸润深度累及肌层、Dukes分期A+B者,差异有统计学意义(P<0.05)。有淋巴结转移、Dukes分期C+D期CD_(44)蛋白表达高于无淋巴结转移和Dukes分期A+B者(P<0.05)。结论 ESA、CD_(44)蛋白表达影响结直肠癌的发生发展,与浸润转移密切相关,在结直肠癌组织检测中同步检测ESA、CD_(44)的蛋白表达,并分析其相关性,有助于结直肠癌的病理判断,在临床中具有一定的临床价值。

宜在蔬菜生产上推广的低毒杀菌剂

1.农抗120。又名抗霉菌素120,可用于防治瓜、果、蔬菜、花卉、烟草、大麦、小麦的白粉病,西瓜和蔬菜的枯萎病,瓜类、水果、蔬菜的炭疽病。一般于发病初期用2％水剂200倍液喷雾,隔10～15天再喷1次。防治瓜类。

热休克蛋白-70在人类糖尿病性白内障晶状体上皮细胞中的表达及意义

目的 :探讨热休克蛋白 70在人类糖性白内障晶状体上皮细胞中的表达及意义。方法 :采用链霉菌抗生物素蛋白碱性磷酸酶 (streptavidin alkalinephosphatase ,S P)免疫组化技术检测热休克蛋白 70 (heatshockprotin 70 ,HSP 70 )在人类糖性白内障 (2 5例 )和正常晶状体 (7例 )上皮细胞中的表达情况。结果 :热休克蛋白 70在人类糖性白内障晶状体上皮中全部为阳性表达 ,而在正常晶状体上皮细胞中全部为阴性表达 ,二者差异有显著性 (χ2 =2 6 .4 2 ,P <0 .0 1)。结论 :热休克蛋白 70在人类糖尿病性白内障发生、发展中起重要作用。

Screening for Streptomyces Hygroscopicus Strains with High Production of Agricultural Antibiotics by Streptomycin Resistance

[Objective] The research aimed to screen Streptomyces hygroscopicus strains with high production of agricultural antibiotics. [ Method] A strain of S. hygroscopicus was screened from the soil of Hainan Island. After natural screening and consecutive ultraviolet induced mutation twice, S6-7 strain was obtained as the original strain then treated by UV irradiation and streptomycin resistance screening, and finally rescreened through shake-flask fermentation. [Result] 7 better strains were selected by primary screening from 62 single colonies which were picked out randomly. After 3 generations of consecutive cultivation on slant media and rescreening, 5 strains presented obvious forward mutation. The forward mutation rate reached 8.06%, and the largest production increasing rate came up to 25.11%. [Conclusion] By combining streptomycin resistance screening and conventional ultraviolet induced mutation, both the antibiotic-producing capacity and forward mutation screening efficiency of the original strain were greatly enhanced.

浅谈紫荆育苗技术

一、种子采集与处理9月至10月荚果由深绿色逐步变成棕黄色时,采集荚果堆放与通风透气的厂棚或晒场里,摊放开来,防止发热霉烂。勤翻薄敞,待有部分荚果开裂时,用工具拍打或用脚踩搓,取出种子,埋于干沙中置阴凉处越冬。二、播种育苗1、苗床处理。选择通风阴凉。

Synthesis and Antibacterial Activities of Erythromycin Derivatives

Aim To synthesize 4″-carbamate derivatives of erythromycin and test their antibacterial activities in vitro. Methods New erythromycin antibacterial agents containing 4″-carbamate group were designed and synthesized from azithromycin via protection, aminoformylation, amination and deprotection. Their antibacterial activities against Staphylococcus aureus strains were tested. Results Nine compounds were synthesized. Their structures were confirmed by MS, IR, ~ 1 H NMR and ~ 13 C NMR, and the synthetic conditi...

Research Progress of Ketolide Antibiotics

Macrolide antibiotics have been widely used in clinic due to their good antibacterial effects and high safety, but the drug-resistant bacteria appear constantly. To solve the problem of drug resistance in pathogens, scholars obtain the third generation of macrolide antibiotics, ketolide antibiotics, which are developed by modifying the structure of macrolide antibiotics, thereby efficiently solving the problem. Ketolide antibiotics are a type of erythromycin derivatives with macrolide structure, and the typical drugs mainly include telithromycin, cethromycin and solithromycin, etc . This paper briefly introduced the recent progress of ketolide antibiotics, with an attempt to provide help to the research and development of new macrolide antibiotics.

非小细胞肺癌凋亡指数与Bax表达的临床病理意义

目的：探讨非小细胞肺癌组织凋亡指数（AI）与Bax表达的临床病理意义。方法：应用免疫组织化学链霉菌抗生物素-过氧化物酶（S-P）法检测46例非小细胞肺癌切除标本以及23例癌旁正常肺组织的Bax表达情况，同时应用末端脱氧核苷酸转移酶介导的dUTP原位切口末端标记（TuNEL）法检测非小细胞肺癌及癌旁正常肺组织AI的大小，并比较不同年龄、不同病理类型、临床分期、不同级别、淋巴结有无转移患者Bax的表达和AI的大小。结果：肺鳞癌与肺腺癌组织中Bax的阳性表达率分别为52．38％、64．00％，低于癌旁肺组织（91．30％）（P〈0．05），不同临床病理特征患者Bax的表达差异无统计学意义（P〉0．05）。肺鳞癌、肺腺癌的AI为（5．67±3．11）％与（6．58±3．58）％，均低于癌旁正常肺组织（14．07±5．16）％（P〈0．05），Ⅲ～Ⅳ期患者AI高于Ⅰ～Ⅱ期患者（P〈0．05），而其它临床病理特征患者AI差异无统计学意义（P〉O．05）。Bax阳性表达的患者AI为（6．93±2．90）％，高于Bax阴性表达的患者（5．04±3．76）％，但差异无统计学意义（P〉0．05）。结论：非小细胞肺癌患者中Bax表达下调、缺失以及AI的下降可能在肿瘤的发生、发展中起一定的作用。

免疫组织化学在乳腺癌鉴别诊断中的临床意义

目的探讨免疫组织化学法在乳腺癌鉴别诊断中的临床意义。方法对江油市人民医院病理科2006年1月至2013年9月收治的852例浸润性乳腺癌组织标本进行链霉菌抗生物素蛋白-过氧化物酶连结法检测。比较不同年龄(≤50岁359例、>50岁493例),不同肿瘤最大直径(T)(T≤2 cm 395例、2 cm<T≤5 cm 401例、T>5 cm 56例),不同临床分期(Ⅰ期251例、Ⅱ期346例、Ⅲ期158例、Ⅳ期97例)的孕激素受体(PR)、雌激素受体(ER)、C-erb B-2、血管内皮生长因子(VEGF)、p53和Ki-67的阳性率。结果PR阳性率在低年龄组(58.77%)、肿瘤T≤2 cm(60.00%)、临床分期早较高且差异均有统计学意义(P<0.05);肿瘤T≤2 cm,ER阳性率(68.35%)显著高于其他肿瘤大小组(P<0.01),不同年龄、临床分期的阳性率差异无统计学意义(P>0.05),C-erb B-2在肿瘤大小、临床分期的阳性率差异均有统计学意义(P<0.05);VEGF、p53和Ki-67在不同年龄组、肿瘤体积组、临床分期的阳性率差异均无统计学意义(均P>0.05)。结论免疫组织化学指标中PR、ER、C-erb B-2在乳腺癌鉴别诊断中具有重要指导意义,可用于临床上鉴别诊断乳腺癌。

湿生扁蕾酮对UC肠纤维化大鼠肠上皮细胞间质转化的影响

目的探讨湿生扁蕾酮对溃疡性结肠炎(UC)肠纤维化大鼠肠上皮细胞间质转化的影响。方法通过2,4,6-三硝基苯磺酸(TNBS)诱导大鼠溃疡性结肠炎肠纤维化模型,利用湿生扁蕾酮干预溃疡性结肠炎肠纤维化模型大鼠,采用链霉菌抗生物素蛋白-过氧化物酶(S-P)法检测上皮表型标记蛋白E-钙黏蛋白(E-cadherin)和间质表型标记蛋白α-平滑肌肌动蛋白(α-SMA)的表达情况,采用实时荧光定量聚合酶链反应(qRT-PCR)法检测E-钙黏蛋白、α-平滑肌肌动蛋白mRNA的表达情况。结果与模型组相比,湿生扁蕾酮显著增加溃疡性结肠炎肠纤维化大鼠肠上皮细胞E-钙黏蛋白的表达,抑制α-平滑肌肌动蛋白的表达。结论湿生扁蕾酮可通过抑制结肠上皮细胞间质转化作用,明显改善2,4,6-三硝基苯磺酸诱导的大鼠溃疡性结肠炎肠纤维化。

IRMA及SP免疫组化染色检测在结肠癌诊断中的意义

目的探讨固相免疫放射分析法(IRMA)及链霉菌抗生物素蛋白-过氧化物酶(SP)免疫组化染色检测在结肠癌诊断中的意义。方法选取在我院手术切除的结肠良性肿瘤患者78例(结肠良性肿瘤组),78例结肠癌患者(结肠癌组),采用IRMA及SP免疫组化染色检测生长抑制因子2(ING2)与基质金属蛋白酶-13(MMP-13)表达水平,判断诊断价值。结果结肠癌组的ING2与MMP-13 IRMA阳性率高于结肠良性肿瘤组(P<0.05);结肠癌组的ING2与MMP-13 SP阳性率高于结肠良性肿瘤组(P<0.05);在156例患者中,IRMA诊断为结肠癌71例,结肠良性肿瘤85例;SP诊断为结肠癌77例,结肠良性肿瘤79例。IRMA诊断结肠癌的敏感性与特异性为89.7%、98.7%,SP诊断的敏感性与特异性为98.7%、100.0%,SP诊断的敏感性高于IRMA(P<0.05)。结论ING2与MMP-13在结肠癌患者IRMA及SP免疫组化染色诊断中都呈现高表达状况,IRMA及SP诊断结直肠癌都有很高的特异性,但是SP诊断的敏感性更好。

Synthesis, Spectral Properties, Antibacterial and Antitumor Activity of Salinomycin Complexes with the Transition Metal Ions Co（ll）, Ni（ll）, Cu（ll） and Zn（ll）

SalNa （sodium salinomycin） reacts with divalent transition metal ions of Co（II）, Ni（II）, Cu（II） and Zn（II） to produce novel compounds characterized by various spectroscopic methods. The interaction of metal （II） ions with SalNa results in the formation of mononuclear complexes of a general composition of [M（Sal）2·（H2O）2] nH2O （n = 0 or 2） where the divalent cations replace Na~ ions from the cavity of initial compound. The new compounds （disalinomycinates） possess an enhanced antibacterial activity against Gram-positive microorganisms as compared to both SalNa and SalH （salinomycinic acid）, respectively. The metal （II） complexes manifest strong concentration dependent cytotoxic effect in experiments using human leukemia cell lines. The complexes of Co0I） and Cu（lI） proved to exert superior activity as compared to the Ni（II） and Zn（II） analogues and are much more cytotoxic than SalNa and SalH. Further studies should be conducted to determine the therapeutic indexes of the new compounds.

Genetic transformation of marine cyanobacterium Synechococcus sp.CC9311(Cyanophyceae) by electroporation

Synechococcus sp.CC9311 is a marine cyanobacterium characterized by type IV chromatic acclimation(CA).A genetic transformation system was developed as a first step to elucidate the molecular mechanism of CA.The results show that Synechococcus sp.CC9311 cells were sensitive to four commonly used antibiotics:ampicillin,kanamycin,spectinomycin,and chloramphenicol.An integrative plasmid to disrupt the putative phycoerythrin lyase gene mpeV,using a kanamycin resistance gene as selectable marker,was constructed by recombinant polymerase chain reaction.The plasmid was then transformed into Synechococcus sp.CC9311 via electroporation.High transformation efficiency was achieved at a field strength of 2 kV/cm.DNA analysis showed that mpeV was fully disrupted following challenge of the transformants with a high concentration of kanamycin.In addition,the transformants that displayed poor growth on agar SN medium could be successfully plated on agarose SN medium.

Genetic Transformation of Nannochloropsis oculata with a Bacterial Phleomycin Resistance Gene as Dominant Selective Marker

The gene ble from Streptoalloteichus hindustanus is widely used as a selective antibiotic marker. It can control the phleomycin resistance, and significantly increase the tolerance of hosts to zeocin. The unicellular marine microalga Nannochloropsis oculata is extremely sensitive to zeocin. We selected ble as the selective marker for the genetic transformation of N. oculata. After the algal cells at a density of 2× 10^7 cells mL-1 was digested with 4% hemicellulase and 2% driselase for 1 h, the protoplasts accounted for 90% of the total. The ble was placed at the downstream of promoter HSP70A-RUBS2 isolated from Chlamydomonas reinhardtii, yielding a recombinant expression construct pMS 188. The construct was transferred into the protoplasts through electroporation （1 kV, 15 μS）. The transformed protoplasts were cultured in fresh f/2 liquid medium, and selected on solid f/2 medium supplemented with 500 ng mL-t zeoein. The PCR result proved that ble existed in the transformants. Three transformants had been cultured for at least 5 generations without losing ble. Southern blotting analysis showed that the ble has been integrated into the genome of N. oculata. The ble will serve as a new dominant selective marker in genetic engineering N. oculata.

血清硅元素水平对矽肺早期诊断意义的探讨

目的探讨血清硅元素水平对矽肺早期诊断的意义。方法采用电感耦合等离子体质谱仪(ICP-MS)检测矽肺模型大鼠血清中硅元素含量,同时采用免疫组织化学-链霉菌抗生物素蛋白-过氧化物酶连结法观察转化生长因子-β1、肿瘤坏死因子-α在大鼠矽肺模型肺组织的表达。结果与对照组比较,大鼠染尘后血清硅元素水平在染尘第一天即见增高,峰值在16 h;其后的7 d、14 d、21 d和28 d,其含量水平亦显著高于对照组,差异有统计学意义。大鼠染尘后肺组织TGF-β1表达与对照组相比差异有统计学意义(P<0.05),表达高峰是染尘后的第7天;TNF-α表达也显著高于对照组,差异有统计学意义(P<0.05)。结论矽肺模型大鼠血清硅元素水平在染尘早期即见明显升高,且早于TGF-β1、TNF-α的表达,提示此一指标在矽肺的早期诊断中具有一定的价值和意义。

Improvement of oxytetracycline production mediated via cooperation of resistance genes in Streptomyces rimosus

Increasing the self-resistance levels of Streptomyces is an effective strategy to improve the production of antibiotics.To increase the oxytetracycline(OTC) production in Streptomyces rimosus,we investigated the cooperative effect of three co-overexpressing OTC resistance genes:one gene encodes a ribosomal protection protein(otrA) and the other two express efflux proteins(otrB and otrC).Results indicated that combinational overexpression of otrA,otrB,and otrC(MKABC) exerted a synergetic effect.OTC production increased by 179%in the recombinant strain compared with that of the wild-type strain M4018.The resistance level to OTC was increased by approximately two-fold relative to the parental strain,thereby indicating that applying the cooperative effect of self-resistance genes is useful to improve OTC production.Furthermore,the previously identified cluster-situated activator OtcR was overexpressed in MKABC in constructing the recombinant strain MKRABC;such strain can produce OTC of approximately7.49 g L^((-1)),which represents an increase of 19%in comparison with that of the OtcR-overexpressing strain alone.Our work showed that the cooperative overexpression of self-resistance genes is a promising strategy to enhance the antibiotics production in Streptomyces.

Preparation and evaluation of liposomal clarithromycin with antimicrobial effect

In this study, we developed a novel liposomal delivery system modified by Tat peptide and wheat germ agglutinin（WGA） with antimicrobial effect. Physicochemical parameters, in vitro antimicrobial, time-kill study, cellular uptake, biofilm formation inhibition and in vivo antibacterial efficacy against Methicillin-resistant Staphylococcus aureus（MRSA） were investigated. Minimum inhibitory concentrations（MICs） and colony-forming units（CFUs） in the time-kill study for Tat-WGA-modified liposomal clarithromycin（CLA-Tat WGALip） were lower than those of free and other modified liposomal CLA. Flow cytometry analysis disclosed that Tat WGALip delivered more coumarin 6 into bacteria. Furthermore, Tat-WGA-modified liposomal CLA efficiently inhibited the formation of MRSA biofiom. CFU of MRSA in the abscess of mice treated with CLA-Tat WGALip was significantly lower than that of any others（P〈0.01）. Collectively, liposomal delivery system modified by Tat and WGA could be a promising anti-resistant infection strategy.

Polydopamine-assisted functionalization of heparin and vancomycin onto microarc-oxidized 3D printed porous Ti6Al4V for improved hemocompatibility,osteogenic and anti-infection potencies

Enhanced antiinfection activities, improved hemocompatibility and osteo-compatibility, and reinforced osseointegration are among the most important considerations in designing multifunctional orthopedic biomaterials.Hereby, anti-infective and osteogenic multifunctional 3 D printed porous Ti6 Al4 V implant with excellent hemocompatibility was successfully designed and fabricated. In brief, osteogenic micro-arc oxidation（MAO） coatings with micro/nanoscale porous topography were generated in situ on3 D printed Ti6 Al4 V scaffolds, on which heparin and vancomycin were easily immobilized. The surface microstructure,morphology, and chemical compositions were characterized employing scanning electron microscopy（SEM）, X-ray photoelectron spectroscopy（XPS） and Fourier transform infrared spectroscopy（FTIR）. High loading capacity and sustained vancomycin release profiles were revealed using high performance liquid chromatography（HPLC）. Favorable antibacterial and antibiofilm performances against pathogenic Staphylococcus aureus（S. aureus） were validated in vitro through microbial viability assays, Live/Dead bacterial staining, and crystal violet staining. Human mesenchymal stem cells（h MSCs） were seeded on the scaffolds and their proliferation and viability were assessed using Cell Counting Kit and Live/Dead cell viability kit. Further, osteoblastic differentiation abilities were evaluated using alkaline phosphatase（ALP） activity as a hall marker. Additionally, the improved hemocompatibility of the heparinized scaffolds was confirmed by activated partial thromboplastin time（APTT）, prothrombin time（PT） and thrombin time（TT）. Overall, our results show that the surface-modified 3 D printed porous Ti6 Al4 V possesses balanced antibacterial and osteogenic functions while exhibiting extra anticlotting effects, boding well for future application in customized functional reconstruction of intricate bone defects.

Selective inactivation of Gram-positive bacteria in vitro and in vivo through metabolic labelling

Bacterial infections are grave threats to human health,particularly those caused by the most common Grampositive bacteria.The massive administration of broad-spectrum antibiotics to treat various bacterial infections has led to the evolution and spread of drug resistance.As a universal antimicrobial technique unapt to induce drug resistance,photothermal therapy(PTT)is attracting extensive attention in recent years.However,its unspecific killing capability and side effects towards adjacent mammalian cells severely impede the practical applications.Herein,we proposed a metabolic engineering strategy to selectively inactivate Gram-positive bacteria by PTT.A bioorthogonal photothermal agent was prepared by the conjugation of IR-780 iodide and dibenzocyclooctyne(IR780-DBCO).Upon pre-metabolizing with 3-azido-D-alanine,Gram-positive bacteria rather than Gramnegative ones,such as Staphylococcus aureus and vancomycinresistant Enterococcus faecalis(VRE),could be specifically tied up by the explosive IR780-DBCO via copper-free click chemistry.Thereafter,they spontaneously detonated under 15 min near-infrared light irradiation and inactivated nearly 100% Gram-positive bacteria in vitro.Moreover,superbug VRE-induced infection was significantly inhibited by this approach in a mouse skin wound model.This metabolic labelling-based photothermal ablation strategy specific to Gram-positive microbes would stimulate the development of precise antibacterial candidates for preclinical applications.

Rapid eradication of antibiotic-resistant bacteria and biofilms by MXene and near-infrared light through photothermal ablation

With the development and rising of antimicrobial resistance,rapid and effective killings of bacteria are urgently needed,especially for antibiotic-resistant bacteria and bacterial biofilms that are usually hard to be treated with conventional antibiotics.Here,a rapid and broad-spectrum antibacterial strategy is demonstrated through photothermal ablation with MXene and light.Ti3C2 MXenes,when combined with 808 nm light,show significant antibacterial effects in just 20 min.The antibacterial strategy is effective to 15 bacterial species tested,including methicillin-resistant Staphylococcus aureus(MRSA)and vancomycin-resistant Enterococci(VRE).In addition,the rapid antibacterial strategy works for MRSA biofilms,by damaging the structures as well as killing bacteria in biofilms.Furthermore,the investigation of the antibacterial mechanisms shows that Ti3C2 with light kills bacteria mainly physically through inserting/contact and photothermal effect.This work broadens the potential applications of MXene and provides a way to eradicate bacteria and biofilms physically,without the likelihood of resistance development.