Insights into sensitizing and eliciting capacity of gastric and gastrointestinal digestion products of shrimp(Penaeus vannamei)proteins in BALB/c mice

Shrimp(Penaeus vannamei)proteins have been shown an allergenic potential;however,little information is available on the sensitizing and eliciting capacity of shrimp protein digestion products.In this study,a BALB/c mice model was used to explore the allergenicity of shrimp protein sample(SPS)and their gastric and gastrointestinal digestion products(GDS/GIDS).As compared with the SPS groups,the GDS/GIDS groups caused lower specific immunoglobulins(Ig E/Ig G1)levels(P<0.05),but higher than the control groups,indicating that the digestion products sensitized the mice.Meanwhile,spleen index,mouse mast cell protease-1(m MCP-1)concentration and proportion of degranulated mast cells were significantly reduced in the GDS/GIDS groups(P<0.05);simultaneously,allergic symptoms,vascular permeability and histopathological changes of tissues were alleviated.Nevertheless,the allergenicity of digestion products cannot be eliminated and still cause systemic allergic reactions in mice.The study showed that the digestion products of shrimp still had high sensitizing and eliciting capacity.

Helicobacter Pylori-induced Gastritis Model in BALB/c Mice Infected With Fresh Isolates from a Human Complex Ulcer Patient

A useful helicobacter pylori-induced gastritis model using BALB/c mice was established for mimicking of human gastritis induced by Helicobacter pylori (H. pylori). The H. pylori isolates were obtained freshly from a human complex ulcer patient. BALB/c mice were fasted for 24 h and then 0.25 mL of 0.2 mol·L -1 NaHCO 3 was administered after by gavage to each mouse and 0.5 mL of 10 9 colonies formation unit per milliliter (CFU/mL) of H. pylori was administered 15 min. On the 3 rd day and 5 th day, the H. pylori inoculations were repeated. The inoculated mice were sacrificed in batch on the 5 th day, in the 2 nd week, 3 rd week and 4 th week. The gastric mucous membrane near pyloric portion was removed, treated and then cultured under microaerobic condition for detection of H.pylori. The remainders of the gastric membrane were fixed by 10% formaldehyde solution for pathological detection. The results showed that the H. pylori could be separated from the gastric membranes of inoculated mice. Obvious invasion of inflammatory cells in the gastric membranes of inoculated mice could be observed from pathological sections. It can be concluded that the inoculating fresh human H. pylori isolates can produce mouse gastritis. This model of BALB/c mice can be used for evaluating the therapeutic agents for the treatment of gastritis induced by H. pylori.

Establishment of BALB/c Mice Model for Food Allergy to Chinese Lobsters

[Objective] This study aimed to establish Balb/c mice model for food allergy caused by Chinese lobsters through using intraperitoneal injection for sensitization and explore methods for in vitro identification and evaluation of food allergy caused by Chinese lobsters. [Method] The 40 male Bal/c mice were divided into ovalbumin（OVA） positive control group, Coca＇s solution negative control group, blank control group and model group. Balb/c mice model was established by intraperitoneally injection of immunized Balb/c mice with OVA or Chinese lobster crude protein with aluminum hydroxide adjuvant. IgE and histamine levels in serum after the second challenge were determined by ELISA method, and the specific IgE antibody titer was determined by passive cutaneous anaphylaxis test（PCA）; additionally, spleen index and histological changes in the small intestine, as well as food allergy symptoms after challenge were also calculated or observed. [Results] After the last challenge, IgE content was（236.75 ±73.39） μg/L in the Chinese lobster crude protein group, revealing no difference with that in the OVA group, but significantly higher than that in either the Coca＇s solution group or the blank control group（P 0.01）;histamine content in serum in the Chinese lobster crude protein group was（406.55±232.79）, significantly higher than that in the blank control group（P0.01）. In the passive cutaneous anaphylaxis test, IgE antibody titer reached 1/16 after the last challenge in the Chinese lobster crude protein group. Spleen index in both Chinese lobster crude protein group and OVA group was significantly greater than that in either the Coca＇s solution group or the blank control group（P0.01）. What＇s more, infiltration of inflammatory cells like lymphocytes and eosnophils at the lamina propria of intestinal mucosa was also observed both Chinese lobster crude protein group and OVA group. [Conclusion] This study established Balb/c mice model for food allergy caused by Chinese lobsters; serum IgE and ELISA assay and specific IgE antibody titer in PCA test can be used for the in vitro identification and evaluation of food allergies caused by Chinese lobsters.

不同基因型牛病毒性腹泻病毒对BALB/c小鼠致病性研究

牛病毒性腹泻病毒(BVDV)是危害养牛业的重要病原。2021年本实验室从患有肺炎的牛肺中分离出1株BVDV(毒株编号为JS2201),经鉴定为1b亚型。为了评价该分离株的毒力与致病性,分别建立了分离株与参考株BVDV-1型Oregon C24V株、BVDV-2型C1602株BVDV感染BALB/c小鼠模型,通过检测攻毒后小鼠临床症状、体重变化、病毒血症情况、血液中白细胞与淋巴细胞的变化、组织器官的病理组织学变化及病毒载量,比较了分离株JS2201与1型和2型参考株的致病性差异。结果显示:BVDV分离株JS2201与参考株均能够成功感染BALB/c小鼠,攻毒BALB/c小鼠均形成病毒血症,导致血液中白细胞数量、淋巴细胞数量以及淋巴细胞百分比短暂下降;BVDV分离株JS2201株攻毒组小鼠肝脏、肾脏、肺脏、脾脏中BVDV载量要高于C24V、C1602株攻毒组;攻毒组小鼠均表现为间质性肺炎病变,JS2201株攻毒组与1型参考株C24V株攻毒组小鼠肺脏病理组织学变化相似,BVDV-2型参考株C1602株攻毒组小鼠肺脏病变更为严重。综上,本研究成功建立了不同亚型BVDV急性感染BALB/c小鼠的模型,且比较分析了分离株与参考株之间的致病性差异,为BVDV疫苗的研制以及致病机制的研究奠定了基础。

面愈贴促进Balb/c小鼠急性面神经损伤后神经功能恢复的作用机制

目的 研究面愈贴促进急性面神经损伤神经功能恢复的作用和可能的机制。方法 随机将120只无特定病原体(SPF)雄性Balb/c小鼠分为假手术组(Sham组,n=40)、模型组(Model组,n=40)和治疗组(M+MYT组,n=40)。模型组和M+MYT组小鼠压榨面神经主干建立急性面神经损伤模型。造模成功后,M+MYT组小鼠给予面愈贴贴敷治疗24 h, Sham组和Model组不做处理。各组每24 h进行面神经功能评分。术后第9天采集脑干面神经核、面神经主干组织和眼球静脉血样本;苏木精-伊红(HE)染色和透射电子显微镜观察面神经主干和面神经核神经元的形态变化;免疫荧光和蛋白质免疫印迹分别检测小鼠面神经核单核细胞趋化蛋白-1(MCP-1)、肿瘤坏死因子-α(TNF-α)、诱导型一氧化氮合酶(iNOS)、内皮型一氧化氮合酶(eNOS)、内皮素-1(ET-1)和血管内皮生长因子(VEGF)表达;酶联免疫吸附试验(ELISA)检测小鼠血清MCP-1、TNF-α、iNOS、eNOS、ET-1和VEGF表达;硝酸还原法检测血清中一氧化氮(NO)浓度。结果 术后第9天,M+MYT组小鼠面神经功能评分低于Model组(P<0.05);HE染色和透射电子显微镜显示,M+MYT组面神经核神经元和神经纤维的损伤较Model组明显改善;免疫荧光、蛋白质免疫印迹和ELISA检测显示,M+MYT组MCP-1、TNF-α、iNOS、ET-1和VEGF表达显著低于Model组(P<0.05),而eNOS表达高于Model组(P<0.05);硝酸还原法显示,NO浓度以M+MYT组最高,Sham组最低(P<0.01)。结论 面愈贴可降低急性面神经损伤后MCP-1、TNF-α、iNOS、ET-1及VEGF表达,增强eNOS和NO表达,有效控制炎症,保护微血管内皮功能,降低血管通透性,促进神经功能恢复。

基于Micro-CT的BALB/c小鼠肺腺瘤动物模型研究

目的 建立基于Micro-CT动态表征的BALB/c小鼠肺腺瘤动物模型研究方法。方法 取80只SPF级雌性BALB/c小鼠,随机分为4组:模型低剂量组(1 mg/g乌拉坦腹腔注射1次)、模型中剂量组(1 mg/g乌拉坦腹腔注射,每周1次,连续2周)、模型高剂量组(1 mg/g乌拉坦腹腔注射,每周1次,连续4周)和空白组(等体积生理盐水腹腔注射)。采用Micro-CT定期监测小鼠肺结节生长情况,Analyze 12.0分析系统绘制小鼠肺部3D图像,苏木素-伊红(HE)染色观察肺组织病理学变化。结果 与空白组相比,各模型组小鼠第11周时均观测到类圆形高密度影的肺结节。结节形成率随造模周数增加而升高,至第21周时,模型高、中、低剂量组结节形成率分别为93.8%、93.8%、87.5%;结节数分别以2~4个、1个、1~2个为主;低剂量组肺结节最大直径平均值高于中、高剂量组(P<0.05);肺结节体积三组之间差异无统计学意义。HE染色结果显示模型高、中、低剂量组病理类型均为肺腺瘤。结论 模型各剂量组均成功诱导肺腺瘤,Micro-CT可对小鼠肺结节生长情况进行表征,其中模型中剂量组结节形成率高,肺腺瘤数量适中,模型稳定,更适合小鼠肺腺瘤动物模型的研究。

美洛昔康对荷瘤BALB/c小鼠的围手术期镇痛效果

目的为了降低肿瘤切除术过程中小鼠的围手术期疼痛,保障动物福利最大化和研究数据的准确性,本研究探究美洛昔康对荷瘤小鼠的围手术期镇痛作用。方法所有BALB/c小鼠均采用舒泰和右美托嘧啶的麻醉方案,分为对照组(A组)和美洛昔康注射组(B组)。采用面部表情评分系统和疼痛行为频率评估小鼠的疼痛程度;监测小鼠的体质量以及脏器的组织病理学方法评估其围手术期安全性。结果B组小鼠的麻醉诱导时间较A组缩短,苏醒时间较A组显著延长(P<0.01);术后3 d内,B组小鼠的面部表情评分和疼痛行为频率较A组显著降低;两组小鼠的主要脏器和肠道组织在组织病理学上均无明显病变,体质量无显著差异,B组小鼠全部存活。结论舒泰和右美托嘧啶麻醉方案联合美洛昔康可以缩短BALB/c小鼠的麻醉诱导时间并使其缓慢苏醒,显著地降低了小鼠的围手术期疼痛,且是安全的。

Immunotoxicity of Acrylamide in Female BALB/c Mice

Objective To investigate the immunotoxicity of acrylamide （ACR） in female BALB/c mice.Methods A total of 200 female mice weighing 18-22 g were randomly divided into four clusters based on body weight, and each weight-based cluster included five groups （10 mice per group）： negative control, positive control （cyclophosphamide）, low, intermediate, and high dose ACR groups, and all the groups were administered ACR by gavage for 30 days. At the end of the study, the immunotoxicological effects of the ACR were evaluated through immunopathology, humoral immunity, cellular immunity, and non-specific immunity. Results The terminal body weight, spleen and thymus weights, lymphocyte counts in the ACR-H group were decreased, pathological changes were observed in lymph glands, thymus and spleen. %T cells in blood lymphocytes were significantly increased in all ACR-treated groups, and a significant reduction of % natural killer（NK） cells and increase of %Th cells were observed in the ACR-H group. interleukin-6（IL-6）, Concanavalin A（ConA）-induced splenocyte proliferation and serum half hemolysis value （HCso） were also significantly suppressed in the ACR-H group. Conclusion ACR elicited an inhibitory effect on cellular and humoral immunity of mice after 30 day feeding.

Molecular Cloning of a Novel cDNA From Mus Muscular BALB/c Mice Encoding Glycosyl Hydrolase Family 1: A Homolog of Human Lactase-Phlorizin Hydrolase

Objective To study the mechanism of lactose intolerance （LI） by cloning the mouse lactase cDNA and recombining a vector. Methods Total murine RNA was isolated from the small intestine of a 4-week-old BALB/c mouse （δ）. Crene-specific primers were designed and synthesized according to the cDNA sequences of lactase-phlorizin hydrolase （LPH） in human, rat, and rabbit. A coding sequence （CDS） fragment was obtained using RT-PCR, and inserted into a clone vector pNEB-193, then the cDNA was sequenced and analyzed using bioinformatics. Results The cDNA from the BALB/c mouse with 912 bp encoding 303 amino acid residues. Analysis of the deduced amino acid sequence using bioinforrnatics revealed that this cDNA shared extensive sequence homology with human LPH containing a conserved glycosyl hydrolase family 1 motif important for regulating lactase intolerance. Conclusion BALB/c mouse LPH cDNA （GenBank accession No： AY751548） provides a necessary foundation for study of the biological function and regulatory mechanism of the lactose intolerance in mice.

Study of primary leiomyosarcoma induced by MNNG in BALB/C nude mice

INTRODUCTIONIt has been well known that MNNG is one of thestrong and multipotential carcinogens that havebeen frequently reported inducing malignant peptictumors.We have successfully induced rat and doggastric adenocarcinomas,squamous cell carcinomasof rat forestomach and gastric leiomyosarcoma

Immunotoxicological Evaluation of Wheat Genetically Modified with TaDREB4 Gene on BALB/c Mice

Objective To evaluate the immunotoxicological effects of genetically modified wheat with TaDREB4 gene in female BALB/c mice. Methods Female mice weighing 18-22 g were divided into five groups （10 mice/group）, which were set as negative control group, common wheat group, parental wheat group, genetically modified wheat group and cyclophosphamide positive control group, respectively. Mice in negative control group and positive control group were fed with AIN93G diet, mice in common wheat group, non-genetically modified parental wheat group and genetically modified wheat group were fed with feedstuffs added corresponding wheat （the proportion is 76%} for 30 days, then body weight, absolute and relative weight of spleen and thymus, white blood cell count, histological examination of immune organ, peripheral blood lymphocytes phenotyping, serum cytokine, serum immunoglobulin, antibody plaque-forming cell, serum half hemolysis value, mitogen-induced splenocyte proliferation, delayed-type hypersensitivity reaction and phagocytic activities of phagocytes were detected. Results No immunotoxicological effects related to the consumption of the genetically modified wheat were observed in BALB/c mice when compared with parental wheat group, common wheat group and negative control group. Conclusion From the immunotoxicological point of view, results from this study demonstrate that genetically modified wheat with TaDREB4 gene is as safe as the parental wheat.

Short-lived AIM2 Inflammasome Activation Relates to Chronic MCMV Infection in BALB/c Mice

Absent in melanoma 2(AIM2)inflammasome is a crucial link bridging the innate host defense and the subsequent adaptive immunity when activated by exogenous double stranded DNA(dsDNA).Through establishing models of disseminated murine cytomegalovirus(MCMV)infection in BALB/c and C57BL/6 mice,we evaluated dynamic expression of AIM2 inflammasome components and its relationship with pathological damage and viral replication,trying tofigure out whether AIM2 inflammasome is related to the chronic mechanism of MCMV.BALB/c and C57BL/6 mice were sacrificed on day 0,1,3,7,14 and 28 post infection.Expression levels of AIM2,pro-caspase-1,caspase-1 p20,pro-IL1β and mature IL1β in primary peritoneal macrophages(PMs)and spleens were detected by Western blotting.Contents of IL18 in the serum were detected by ELISA.Pathological examinations of livers were performed,and mRNA levels of MCMV glycoprotein B(gB)in salivary glands also assessed.Results showed that expression levels of AIM2 in PMs and spleens of C57BL/6 mice increased on day 3,even continued to day 28;caspase-1 p20 and mature IL1β increased on day 7,14 and 28;the persistently high expression of IL1β in the serum started on day 1,showing a double peak curve.As for BALB/c mice,expression of AIM2 in PMs increased on day 1 and day 7,while contents of AIM2 in spleens increased on day 1 and day 3;caspase-1 p20 and mature ILip merely increased 7 days fter infection.Thereafter,expression levels of AIM2,caspase-1 p20,mature IL1β and IL18 were limited;the duration of AIM2 inflammasome activation in BALB/c mice was much shorter than that in C57BL/6 mice.The severer pathological damage and more viral replications in BALB/c mice further proved the deficient antiviral immunity to MCMV.In conclusion,the activation of AIM2 inflammasome in BALB/c mice was short-lived,which is quite possibly related to the chronicity of MCMV infection.

腹腔注射攻毒建立蜱媒脑炎病毒感染BALB/c小鼠模型

目的通过腹腔注射攻毒途径建立蜱媒脑炎病毒(TBEV)感染BALB/c小鼠模型,观察小鼠的感染症状和病毒在小鼠体内的复制特征。方法将6周龄雌性BALB/c小鼠随机分为未感染病毒对照组、1×10^(3)空斑形成单位(PFU)TBEV感染组(以每只小鼠1×10^(3)PFU的攻毒剂量经腹腔注射感染小鼠)、1×10^(4)PFU TBEV感染组(以每只小鼠1×10^(4)PFU的攻毒剂量经腹腔注射感染小鼠),观察小鼠的感染症状、体重变化与生存情况。通过H-E染色观察小鼠脑、脾的病理改变,采用免疫组织化学染色检测小鼠脑、脾中TBEV蛋白的表达,采用空斑试验检测小鼠脑、脾中TBEV滴度的动态变化。结果1×10^(4)PFU TBEV感染组小鼠于感染后第6天出现弓背、后肢瘫痪等感染症状,1×10^(3)PFU TBEV感染组小鼠于感染后第7天出现上述症状。与未感染病毒对照组小鼠相比,1×10^(4)PFU TBEV感染组小鼠从第5天、1×10^(3)PFU TBEV感染组小鼠从第6天开始体重明显降低,差异有统计学意义(P<0.05)。1×10^(4)PFU TBEV感染组小鼠第7天开始死亡、第8天全部死亡,1×10^(3)PFU TBEV感染组小鼠第7天开始死亡、第9天全部死亡。H-E染色结果显示,TBEV感染后第5天、第7天小鼠大脑皮质仅出现少量神经元核固缩、深染,未观察到炎症细胞浸润;TBEV感染后第5天小鼠脾红髓轻度淤血、多见中性粒细胞,第7天中性粒细胞数量增多。免疫组织化学染色结果显示,TBEV感染后第5天小鼠脑与脾少量表达TBEV蛋白,第7天TBEV蛋白表达量增加。1×10^(3)PFU TBEV感染第7天,小鼠脑内TBEV滴度高达(1.3±0.6)×10^(5)PFU/mL,而脾中TBEV滴度为(1.3±0.6)×10^(3)PFU/mL。结论经腹腔注射攻毒成功建立了TBEV感染BALB/c小鼠模型,TBEV可在小鼠体内增殖并具致病性。

Cyclosporin A protects Balb/c mice from liver damage induced by superantigen SEB and D-GaIN

AIM To investigate the pathogenic effect ofSEB and D-GalN on liver and the protection ofcyclosporin A, the relationship between hepaticapoptosis and necrosis and the possiblemechanism of acute hepatic necrosis.METHODS After staphylococcal enterotoxin B(SEB ) mixed with D--galactosamine (D-GaiN )were injected intraperitoneally into Balb/c miceand those previously treated with cyclosporin A,blood samples were collected and livers wereisolated at 2, 6, 12 and 24 h. Patterns othepatocellular death were studiedmorphologically and biochemically, circulatingcytokines (TNF-a, IFN--y ) and mice mortalitywithin 24h was assessed.RESU’LTS The SEB could induce the typicalapoptotic changes of hepatocytes, the D-GaiNcould induce hepatocytes apoptosis anddegeneration at the same time, and the micehaving received the SEB + D-GaiN injectionsdeveloped apoptosis at 2 and 6 h, but after 12 hhepatocytes were characterized by severein jury, whereas all the examinations in thecyclosporin A treated mice were normal.CONCLUSION Hepatic cell apoptosis might berelated to necrosis, and massive hepatocyteapoptosis is likely the initiating step of acutehepatic necrosis in mice. The effects induced bySEB and D--GaiN on hepatocytes might bemediated by T cells, and could be prevented bycyclosporin A.

A murine model of dengue virus infection in suckling C57BL/6 and BALB/c mice

Dengue is a significant public health concern across tropical and subtropical regions worldwide,principally causing disease in children.Very young children are at increased risk of severe manifestations of dengue infection.The mechanism of dengue disease in this population is not fully understood.In this study,we present a murine model of dengue virus primary infection in suckling C57BL/6 and BALB/c mice in order to investigate disease pathogenesis.Three-day-old C57BL/6 mice intraperitoneally infected with DENV-2 NGC were more susceptible to infection than BALB/c mice,showing increased liver enzymes,extended viremia,dissemination to organs and histological alterations in liver and small intestine.Furthermore,the immune response in DENV-infected C57BL/6 mice exhibited a marked Th1 bias compared to BALB/c mice.These findings highlight the possibility of establishing an immunocompetent mouse model of DENV-2 infection in suckling mice that reproduces certain signs of disease observed in humans and that could be used to further study agerelated mechanisms of dengue pathogenesis.

Assessment of immune responses and intestinal flora in BALB/c mice model of wheat food allergy via different sensitization methods

Increasing incidences showed that food allergies have attracted more and more attention from researchers.BALB/c mice were sensitized with wheat gluten combined with aluminum hydroxide adjuvant via intraperitoneal injection,transdermal sensitization,and oral gavage sensitization route.Results showed that all the three sensitization methods could induce allergic symptoms;increase the serum antibody(total immunoglobulin E(IgE),specific IgE,IgG,IgA)and histamine content;promote the secretion of Th2 cytokines(interleukin(IL)-4,IL-5,IL-13)and inflammatory factors(IL-6,IL-17 A,IL-10);and inhibit the production of Th1 cytokines(IFN-γ,IL-2).However,the allergic symptoms of mice sensitized by intraperitoneal injection were the most obvious among the three models.The level of serum antibodies in intraperitoneal injection group was significantly higher than control.Subsequently,16 S rRNA sequencing technology was used to analyze the intestinal flora of mice.The results showed that the abundance of Firmicutes in the wheat protein sensitized group was lower than that in the normal group,while the abundance of Bacteroidetes was higher,and Lactobacillus was the difference marker in normal group.Bacterial species diversity analysis showed that the species richness and diversity of intestinal flora in mice were decreased,the difference between mice induced by intraperitoneal injection and normal control group mice was the most significant.Taken together,these results show that among three sensitization methods used to build a mouse model with aluminum hydroxide as adjuvant,intraperitoneal injection is the comparably best way to build a mouse sensitization mode.

BALB/cSpf小鼠和野生型BALB/c小鼠生物学特性比较

目的比较分析BALB/cSpf小鼠与野生型BALB/c(N-BALB/c,N:normal)小鼠生物学特性指标差异,为其实验应用提供基础数据。方法3~12周,每周对雌雄BALB/cSpf小鼠和N-BALB/c小鼠生长曲线、采食量、饮水量进行统计分析;取BALB/cSpf小鼠雌性和N-BALB/c小鼠雌性各30只,二雌一雄长期同居交配方式繁殖饲养,记录产仔数,产仔性别;采集血液进行17项血液生化指标和22项血液生理指标测定。结果BALB/cSpf小鼠和N-BALB/c小鼠采食量和饮水量差异不显著(P>0.05),BALB/cSpf小鼠体质量显著高于N-BALB/c小鼠(雌性从第5周开始,雄性从第3周开始)。1~3胎产仔数及产仔性别差异不显著。雌性BALB/cSpf小鼠生理指标WBC、Neu、Lym、RBC、HGB、HCT、MPV、PDW显著高于N-BALB/c小鼠(P<0.01)。雄性BALB/cSpf小鼠生理指标WBC、Neu、Lym、Mon、HCT、MCHC显著低于N-BALB/c小鼠(P<0.01)。雌性BALB/cSpf小鼠生化指标UREA显著高于N-BALB/c小鼠(P<0.05);AST、CK、LDH显著低于N-BALB/c小鼠(P<0.01)。BALB/cSpf小鼠生化指标TG、TC、LIP、UREA、MgⅡ显著高于N-BALB/c小鼠(P<0.01);ALT、AST、Glu-G、Ca显著低于N-BALB/c小鼠(P<0.01)。结论BALB/cSpf小鼠和N-BALB/c小鼠生物学特性有一定差异。

Acupuncture enhances anticancer effects of cyclophosphamide on 4T1 tumors via suppression of angiogenesis in BALB/c mice

Objective:Acupuncture is considered an important part of the alternative medical treatment of tumors.However,it is still unclear whether acupuncture has a direct effect on inhibition of tumor growth.The purpose of this study was to evaluate the effects of acupuncture treatment on tumors in BALB/c mice that were or were not administered cyclophosphamide.Methods:A 4T1 breast carcinoma mouse model for this study was established.When the mice developed 5 mm × 5 mm visible subcutaneous tumors,they were subjected to acupuncture and/or cyclophosphamide (CTX) treatment for 15 days.Results:Acupuncture and CTX treatment both reduced the size of the solid tumors.When used in conjunction,the tumor size reduction was even more obvious.Hematoxylin and eosin staining showed that acupuncture had a significant effect on induction of tumor cell necrosis and inhibition of angiogenesis.Immunohistochemistry showed that the expression of CD34 and matrix metalloproteinase-9 (MMP-9),which were related to angiogenesis,decreased after treatment with either acupuncture or CTX.However,when the two treatments were used in conjunction,they showed a synergistic effect on inhibiting the expression of CD34.However,the synergistic effect was not observed onMvMP-9 expression.Conclusion:Acupuncture plays an important role in improving the effect of chemotherapy by inhibiting angiogenesis.

Synergetic effect of Egyptian propolis in immunization of BALB/c mice against bovine cysticercosis

Objective: To evaluate the synergetic effect of an ethanolic extract of Egyptian propolis in immunization of BALB/c mice with Taenia saginata(T. saginata) crude antigen against bovine cysticercosis, with reference to its effects on liver and kidney functions.Methods: Sixty female mice BALB/c strain weighing 20 to 25 g and 6-8 weeks old were randomly allocated into six groups of ten mice each. Mice in groups 1 and 2(G1 and G2) were immunized intraperitoneally with 100 μg of T. saginata crude antigen in 100 μL phosphate buffer saline emulsified in Freund's adjuvant. Besides, the mice in G2 were administered with propolis extract simultaneously with immunization. Control mice were either administered with propolis extract(G3) or injected with the same volume of phosphate buffer saline emulsified in Freund's adjuvant(G4). The mice in G5 were non-immunized infected control while, those in G6 were non-immunized non-infected control. Two weeks after the last immunization, each mouse was challenged intraperitoneally with 5 000 oncospheres except those of G6. Ethanolic extract of propolis was prepared at a dose 50 mg/kg body weight.Results: After 24 weeks of challenge, the mice in G2 showed the highest level of protection(100%), with no cyst being detected rather than mice in G1(33.3% protection). Additionally,the ELISA results, in this study, showed higher antibody titer in G2 with reduction the alteration in liver and kidney functions compared to G1.Conclusions: Egyptian propolis could increase the level of protection against experimental challenge infection with T. saginata eggs when administered simultaneously with immunization. Furthermore, it could enhance the production of antibodies to immunized antigen and decrease the alteration in liver and kidney functions.

Adjuvant activity of Pasteurella multocida A strain,Pasteurella multocida B strain and Salmonella typhimurium bacterial DNA on cellular and humoral immunity responses against Pasteurella multocida specific strain infections in Balb/c mice

Objective: To evaluate the effects of Pasteurella multocida(P. multocida) vaccines on the expression and release of antibodies, interleukin(IL)-6 and IL-12 by serum. Methods: Balb/c mice were immunized with two formalin and iron inactivated vaccine doses within 2 weeks. The vaccines were adjuvant with P. multocida A strain, P. multocida B strain and Salmonella typhimurium bacterial DNA(AbDNA, BbDNA and SbDNA for short, respectively). The animals were challenged 4 weeks after immunization. Blood of mice was collected to detect the change of specific antibody, IL-6, and IL-12 using ELISA. Results: The specific antibody and interleukins in the immunized group increased significantly compared to the control mice after vaccination and challenge(P<0.05). The highest release of these cytokines was obtained by P.multocida inactivated with iron and adjuvant with AbDNA at a concentration of 25 μg/mL. The antibody titer peak was 0.447 in mice vaccinated with iron-killed whole-cell antigen adjunct with AbDNA. The time-courses of release showed that bacterial DNA was able to stimulate IL-6 and IL-12 production more than alum(P<0.05). Conclusions: Our findings introduce that bacterial DNA is capable of releasing an immunological response with several cytokines.These indicate that bacterial DNA entrapped with killed P. multocida antigen is a new and effective adjuvant to enhance specific immunity and resistance of animal against the infectious pathogen, which could simplify the development of highly promising strong adjuvant.