灭活的双歧杆菌治疗菌群失调致小鼠肠源性感染

目的:观察灭活的双歧杆菌对菌群失调致小鼠肠源性感染的治疗作用.方法:40只昆明种小鼠随机分为4个处理组:活菌组、死菌组、自然恢复组、正常对照组,每组10只.对血清总蛋白(TP)、血清白蛋白(ALB)、血清球蛋白(GLO)、γ-谷氨酰氨转肽酶(GGT)、血清尿素氮(BUN)、乳酸脱氢酶(LDH)、血肌酐(CRE)以及组织匀浆内的示踪菌进行测定.结果:死菌组与自然恢复组比较,TP、GLO明显升高(71.46±2.01g/Lvs64.61±2.16g/L;35.63±1.57g/Lvs330.73±1.03g/L,P<0.05),GGT、BUN则显著降低(2.47±0.28IU/Lvs7.13±1.19IU/L;8.32±0.71mmol/Lvs12.05±0.64mmol/L,P<0.01).死菌组与活菌组、正常对照组比较,LDH明显降低(2561.23±61.40IU/Lvs2951.70±155.61IU/L,2895.27±104.38IU/L,P<0.05).死菌组与自然恢复组比较,肾、肝、肺组织匀浆内肠杆菌数量均明显下降(1.19±0.37,1.56±0.20,1.62±0.15vs2.17±0.97,2.42±0.14,2.20±0.09,P<0.05或P<0.01).结论:双歧杆菌灭活菌悬液具有与活菌悬液相同的抗菌群失调致小鼠肠源性感染作用.     

Associations between Plasma Folate and Vitamin B12, Blood Lead,and Bone Mineral Density among Adults and Elderly Who Received a Health Examination

This study hypothesized that plasma folate and vitamin B₁₂ levels modified the association between blood lead and cadmium and total urinary arsenic levels and bone loss. A total of 447 study subjects who received a physical examination at the Wanfang Hospital Medical Center were recruited. Bone loss was defined as a calcaneus bone mineral density T-score less than −1. Blood cadmium and lead concentrations were measured by ICP-MS. Urinary arsenic species were determined using HPLC-HG-AAS. A SimulTRAC-SNB radioassay was used to measure plasma folate, vitamin B₁₂, and homocysteine levels. Total urinary arsenic and blood lead concentration were positively correlated with the odds ratio (OR) for bone loss in a dose–response manner. The OR and 95% confidence interval (CI) for bone loss in participants with blood lead concentrations > 56.14 versus ≤33.82 μg/dL were 1.82 and 1.10–3.01. No correlation between plasma folate and vitamin B₁₂ levels alone and bone loss was observed. However, this study is the first observational study to find that blood lead concentrations tend to increase the OR of bone loss in a low plasma folate and plasma vitamin B₁₂ group with multivariate ORs (95% CI) of 2.44 (0.85–6.96).     

可移植性人脑胶质瘤组织裸小鼠脑内移植及其MR显像研究

背景与目的:文献报道的胶质瘤动物原位移植模型,大多数是将细胞悬液立体定向接种于鼠脑内,操作繁杂,耗时长,难以在短时间内完成批量实验.本研究接种可移植性人脑胶质瘤组织于裸小鼠脑内,探讨建立人脑胶质瘤裸小鼠原位移植模型及其MR活体成像的可行性.方法:在套管针内,置入位于裸小鼠皮下生长的可移植性人脑胶质瘤组织2 mm3,经微型颅钻钻颅孔后经套管针推入裸小鼠右尾状核内,第30天在配有小鼠专用micro-23微线圈的1.5T MR机上进行头颅扫描和专用软件测量显像的肿瘤体积.继而取全脑作连续冰冻切片,HE染色,在光学显微镜下观察肿瘤病理特征,在体视显微镜目镜下用测微尺测量肿瘤.然后将上述两种方法测得的数据进行统计学分析,评价荷瘤鼠活体MR显像计算肿瘤体积的可行性.结果:经MR扫描的15只颅内接种肿瘤的小鼠,有14只的尾状核区域见到肿瘤显像;在脑切片上,与MR像相同部位见到了肿瘤组织.经脑切片测得的肿瘤体积[(23.19±10.18)mm3]和经MRI测得的肿瘤体积[(23.45±11.64)mm3]比较差异无统计学意义(P＞0.05).肿瘤模型制作成功率为93%(14/15),肿瘤模型MR显像成功率为100%(14/14).结论:经套管针定量植入胶质瘤组织于裸小鼠尾状核,制作可移植性裸小鼠人脑胶质瘤原位模型,具有操作便捷、省时、便于批量制作、致瘤率高等优点.配有小鼠专用微型线圈的1.5T MR机可用于荷瘤裸小鼠的头颅成像、在活体上对移植瘤进行定位和体积测量等研究.     

Pathogenicty and immune prophylaxis of cag pathogenicity island gene knockout homogenic mutants

AIM: To clarify the role of cag pathogenicity island (cagPAI) of Helicobacter pylori (H pylori ) in the pathogenicity and immune prophylaxis of H pylori infection. METHODS: Three pairs of H pylori including 3 strains of cagPAI positive wildtype bacteria and their cagPAI knockout homogenic mutants were utilized. H pylori binding to the gastric epithelial cells was analyzed by flow cytometry assays. Apoptosis of gastric epithelial cells induced by H pylori was determined by ELISA assay. Prophylaxis effect of the wildtype and mutant strains was compared by immunization with the sonicate of the bacteria into mice model. RESULTS: No difference was found in the apoptasis between cagPAI positive and knockout H pylori strains in respective of the ability in the binding to gastric epithelial cells as well as the induction of apoptosis. Both types of the bacteria were able to protect the mice from the infection of H pylori after immunization, with no difference between them regarding to the protection rate as well as the stimulation of the proliferation of splenocytes of the mice. CONCLUSION: The role of cagPAI in the pathogenicity and prophylaxis of H pylori infection remains to be cleared.     

Expression of Helicobacter pylori AlpA protein and its immunogenicity

AIM: To construct a recombinant strain which expresses adhesin AlpA of Helicobacter pylori (H pylori) and to study the immunogenicity of adhesin AlpA. METHODS: Gene Ab, which was amplified from H pylori chromosomal DNA by PCR technique, was sequenced and the biological information was analyzed, and inserted into the Nco I and Not I restriction fragments of the expression vector pET-22b(+) using T4 DNA ligase. The resulting plasmid pET-AlpA was transformed into competent E.coli BL21(DE3) cells using ampicillin resistance for selection. Recombinant strains were incubated in 5 mL LB with 100 μg/mL ampicillin overnight at 37 ℃. Sonication of BL21(DE3)pET-22b(+)/AlpA was analyzed by Western blot to detect AlpA immunogenicity. RESULTS: The gene encoding AlpA protein was amplified by PCR with chromosomal DNA of H pylori Sydney strain (SS1) as templates. It revealed that AlpA DNA fragment amplified by PCR had approximately 1 500 nucleotides, compatible with the previous reports. The recombinant plasmid pET-22b(+)/AB was successfully constructed. DNA sequencing showed one open reading frame with the length of 588 bp. It encoded seven conservative regions that showed good antigenicity and hydrophobicity by Parker and Welling method. Furthermore, INTERNET EXPASY, NNPREDICT and ISREC predicted that it was a porin-like structure consisting of β-pleated sheets that were embedded in the outer membrane. BLAST analyzed 836 767 protein sequences and found that the similar sequences were all belonging to H pylori OMP sequences. SDS-PAGE and scan analysis showed that the molecular weight of AB was 22.5 ku and recombinant protein amounted to 29% of the total bacterial protein, among which dissolved expression amounted to 21.9% of sonicated supernatant. The rAB purity amounted to 96% through affinity chromatography. Western blot analysis of rAB confirmed that it could be specially recognized by serum form rabbit immunized with AlpA and H pylori infected. CONCLUSION: Adhesin AlpA recombinant protein may be a potential vaccine for control and treatment of H pylori infection.     

布地奈德与泼尼松分别联合硫唑嘌呤治疗自身免疫性肝炎

目的比较布地奈德联合硫唑嘌呤与泼尼松联合硫唑嘌呤治疗自身免疫性肝炎(AIH)的疗效。方法选择宝鸡市中心医院2015年1月至2018年1月收治的90例AIH患者,随机分为布地奈德组联合硫唑嘌呤与泼尼松组联合硫唑嘌呤,各45例。2组均连续治疗2年。比较2组临床总缓解率、治疗前后的血清ALT、AST、ALP、γ-GT水平与不良反应总发生率。结果布地奈德组临床总缓解率88.89%(40/45),显著高于泼尼松组71.11%(32/45)(P<0.05)。2组治疗2年后的血清ALT、AST、ALP、γ-GT水平均较治疗前显著下降(P<0.05)。布地奈德组治疗后的血清ALT、AST、ALP、γ-GT水平分别为(27.42±5.36)U/L、(29.97±4.52)U/L、(88.11±14.32)U/L、(49.52±7.62)U/L显著低于泼尼松组(P<0.05)。布地奈德组治疗期间不良反应总发生率15.56%(7/45),显著低于泼尼松组35.56%(16/45)(P<0.05)。结论布地奈德联合硫唑嘌呤治疗AIH可获得更高的缓解率,更明显的生物化学指标改善,且不良反应更少,效果优于泼尼松联合硫唑嘌呤。     

大鼠急性肝功能衰竭模型的建立及凝血功能的变化

目的：观察不同剂量D-氨基半乳糖（D-GalN）联合脂多糖（LPS）对大鼠急性肝衰竭模型的影响及凝血功能的变化,建立理想的急性肝衰竭大鼠模型。方法：SD大鼠随机分为正常组, D-GalN高、中、低剂量组,每组10只,除正常组外,其余各组注射不同剂量D-GalN联合LPS建立急性肝衰竭大鼠模型,观察大鼠的死亡率,检测大鼠0、12、24、48、72 h肝功能及凝血功能的水平;HE染色,观察肝脏病理的变化。结果：D-GalN高、中、低剂量组72 h内的死亡率分别为：60%、30%、10%;不同剂量组不同时间点血液中丙氨酸氨基转移酶（ALT）、天门冬氨酸氨基转移酶（AST）、总胆红素（TBIL）、凝血酶原时间（PT）、国际标准比率（INR）、血浆纤维蛋白原（FIB）含量与正常组比较均有明显差异（P<0.05）;但在高、中、低剂量组间比较,ALT、AST无统计学意义,TBIL、PT、INR、FIB均有统计学差异（P<0.05）。结论：凝血功能在肝衰竭模型建立中为更加稳定的检测指标,通过肝功能、凝血功能、病理形态综合诊断,中剂量D-GalN联合LPS腹腔注射48 h后建立的急性肝衰竭大鼠模型与临床肝衰竭症状比较一致,中剂量是理想的造模剂量。     

电针内关对缺血再灌注损伤大鼠心肌肌浆网钙泵活性及mRNA表达的影响

目的：观察电针内关对缺血再灌注损伤大鼠心肌SERCA活性及其基因表达的影响,并以神门穴、合谷穴作对比研究。方法：实验分为假手术组、心肌缺血再灌注模型组（模型组）、电针内关组、电针神门组、电针合谷组,用无机磷比色法测定SERCA活性、RT-PCR法分析SERCA基因表达。结果：与假手术组比,模型组SERCA活性及其基因表达均有非常显著性差异（P〈0.01）,电针内关组、电针神门组与模型组比较差异有显著性（P〈0.05,P〈0.01）,电针内关组优于电针神门组（P〈0.05,P〈0.01）。结论：电针内关上调心肌SERCA基因表达,增强SERCA活性是实现对再灌注损伤心肌组织保护作用的途径之一。电针内关与电针神门两组疗效差异说明经穴对靶器官机制调节作用与经脉（穴）脏腑间的特异联系密不可分。     

Retracted Article: Exosome-derived PTENP1 suppresses cisplatin resistance of bladder cancer (BC) by suppressing cell proliferation,migration and inducing apoptosis via the miR-103a/PDCD4 axis

Bladder cancer (BC) is a lethal cancer that threatens the health of millions of people. Chemotherapy drug resistance, for example, cisplatin (DDP) resistance, is a huge limitation for BC therapy. PTEN pseudogene-1 (PTENP1) has been identified as a significant biomarker of multiple cancers. Therefore, it is essential to illuminate the molecular mechanism of PTENP1 in BC cell DDP resistance and progression. Serum exosomes were isolated using an ExoQuick precipitation kit. Serum exosomes were round-shaped vesicles of 100 ± 60 nm in size. The expression of PTENP1 was down-regulated in serum exosomes isolated from cisplatin non-responsive patients compared with responsive patients. ROC curves certified the diagnostic value of PTENP1. Apparently, PTENP1 transfection inhibited DDP-resistant BC cell proliferation, migration, cisplatin resistance and facilitated apoptosis. Next, we discovered that PTENP1 was a sponge of miR-103a, while PDCD4 was a target of miR-103a. More importantly, PTENP1 regulated DDP-resistant cell viability, migration, apoptosis and cisplatin resistance by interacting with the miR-103a/PDCD4 axis. In addition, PTENP1 hindered tumor growth of cisplatin-resistant mice. Exosome-derived PTENP1 suppressed the DDP resistance of BC by inhibiting cell proliferation, migration and promoting apoptosis through regulating the miR-103a/PDCD4 axis, representing a targeted therapy for DDP-resistant BC patients.

Bladder cancer (BC) is a lethal cancer that threatens the health of millions of people.