Helicobacter pylori (Hp) is one of most common pathogens causing gastrointestinal disorder including gastric ulcer, duodenal ulcer and gastric cancer, etc. It has been verified as class I carcinogen by WHO. Nowadays, combination antibiotics and proton pump inhibitor are mainly used to erase Hp in clinical application. However, with the increased resistance of Hp, the vaccine against Hp might become the best strategy to eradicate Hp. Elements including urease, virulence factor, outer membrane protein, flagella, play an important role in Hp infection, colonization and reproduction. They have become potential candidate antigens in the development of Hp vaccine, as reported in previous studies. Presently, these antigens-centric vaccines have been tested in animal models. Therefore, this article reviews the studies on Hp vaccine with urease, virulence genes, outer membrane protein and flagella as their candidate antigens, in an attempt to provide insights for research in this regard.
Animals ; Helicobacter pylori ; Urease/genetics* ; Helicobacter Infections/prevention & control* ; Vaccines ; Membrane Proteins

ObjectiveBased on ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS^E） technique， we identified qualitatively the metabolites of aristolochic acid（AAs） in rat in order to analyze the metabolic differences between water extract of Aristolochiae fructus（AFE） and Aristolochic acid Ⅰ（AAⅠ）. MethodSD rats were selected and administered AFE（110 g·kg^-1·d^-1） or AAⅠ（5 mg·kg^-1·d^-1） by oral for 5 days， respectively. Serum， urine and feces were collected after administration. Through sample pretreatment， ACQUITY UPLC BEH C₁₈ column（2.1 mm×100 mm， 1.7 μm） was used with the mobile phase of 0.01% formic acid methanol（A）-0.01% formic acid water（B， containing 5 mmol·L^-1 ammonium acetate） for gradient elution（0-1 min， 10%B； 1-7 min， 10%-75%B； 7-7.2 min， 75%-95%B； 7.2-10.2 min， 95%B； 10.2-10.3 min， 95%-10%B； 10.3-12 min， 10%B） at a flow rate of 0.3 mL·min^-1. Positive ion mode of electrospray ionization（ESI⁺） was performed in the scanning range of m/z 100-1 200. In combination with UNIFI 1.9.4.053 system， the Pathway-MS^E was used to qualitatively analyze and identify the AAs prototype and related metabolites in biological samples（serum， urine and feces）， and to compare the similarities and differences of metabolites in rats in the subacute toxicity test between AFE group and AAⅠ group. ResultCompared with AAⅠ group， 6， 10， 13 common metabolites and 14， 20， 30 unique metabolites were identified in biological samples（serum， urine and feces） of AFE group， respectively. Moreover， the main AAs components always followed the metabolic processes of demethylation， nitrate reduction and conjugation. Compared with common metabolites in AAⅠ group， prototype components of AAⅠ in serum and most metabolic derivatives of AAⅠ［AAⅠa， aristolochic lactam Ⅰ（ALⅠ）a， 7-OHALⅠ and its conjugated derivatives］ in biological samples were significantly increased in AFE group（P<0.05， P<0.01）， except that the metabolic amount of ALⅠ in feces of AFE group was remarkably lowed than that of AAⅠ group（P<0.01）. In addition， a variety of special ALⅠ efflux derivatives were also identified in the urine and feces of the AFE group. ConclusionAlthough major AAs components in AFE all show similar metabolic rules as AAⅠ components in vivo， the coexistence of multiple AAs components in Aristolochiae Fructus may affect the metabolism of AAⅠ， and achieve the attenuating effect by increasing the metabolic effection of AAⅠ and ALⅠ.

[摘 要] 目的：体内外实验探讨木鳖子单体化合物对羟基桂皮醛[Momordica cochinchinensis(Lour.)Spreng.p-hydroxycinnamaldehyde，CMSP]对小鼠黑色素瘤移植瘤生长和转移的影响及其作用机制。方法：建立荷瘤小鼠动物模型，并将18只C57BL/6小鼠随机分成3组（每组6只）：对照组（腹腔注射0.1 ml生理盐水）、CMSP治疗组（分别腹腔注射0.1 ml 1、2 mg/ml CMSP），给药的第5天开始，每次给药前用卡尺分别测量和计算小鼠移植瘤的体积，实验结束后称量移植瘤的质量；H-E染色后光镜观察肝组织的病理学变化；免疫组织化学SP法观察移植瘤组织E-cadherin和vimentin蛋白的表达。采用细胞划痕和Transwell实验分别检测CMSP实验组（10、20 µg/ml）黑色素瘤B16细胞24、48 h的迁移能力，qPCR法检测CMSP处理24 h后B16细胞EMT相关mRNA表达，WB法检测CMSP处理B16细胞48 h后β-catenin、p-β-catenin（Ser675）、vimentin和E-cadherin蛋白的表达水平。结果：CMSP治疗组小鼠移植瘤平均体积和肿瘤质量明显降低（均P<0.05）；对照组小鼠肝脏中转移灶的数量明显多于CMSP（1、2 mg/kg）治疗组（均P<0.05），CMSP（2 mg/kg）处理组小鼠的肝组织内未发现明显转移灶。CMSP治疗组（1、2 mg/kg）移植瘤组织中E-cadherin蛋白表达水平明显高于对照组（均P<0.05），而vimentin蛋白表达显著低于对照组（均P<0.01）。体外实验中，CMSP实验组（10、20 μg/ml）B16细胞24、48 h后划痕愈合率较对照组均明显降低（均P<0.05）。20 μg/ml CMSP处理B16细胞24、48 h后穿过Transwell小室的细胞数较对照组则显著下降（均P<0.01）。CMSP（10、20 μg/ml）处理B16细胞后β-catenin mRNA表达水平较对照组明显降低（均P<0.01），E-cadherin mRNA表达水平则明显升高（均P<0.05），而vimentin mRNA表达水平在10 μg/ml处理组与对照组相比差异无统计学意义（P>0.05），20 μg/ml处理组则明显降低（P<0.01）。与对照组相比，CMSP实验组（10、20 μg/ml）处理B16细胞后β-catenin、p-β-catenin和vimentin蛋白表达均显著降低（均P<0.01），而E-cadherin蛋白表达则明显升高（均P<0.01）。结论：CMSP能够抑制小鼠黑色素瘤移植瘤的生长和转移，其作用机制可能与抑制wnt/β-catenin通路的活性相关。

Anti-contactin associated protein-like 2 (CASPR2) antibody encephalitis is a rare autoimmune encephalitis with variable clinical symptoms and atypical imaging manifestations. The prognosis of the patients with severe disease is poor. Reversible posterior leukoencephalopathy syndrome is rarely reported in autoimmune encephalitis. The clinical data, diagnosis and treatment of a patient with anti-CASPR2 antibody encephalitis complicated with reversible posterior encephalopathy syndrome were reported, in order to improve the understanding of clinicians on the rare disease complicated with atypical imaging manifestations.

Objective:To analyze the expression pattern, mechanism and clinical significance of melanoma-associated antigen-C2 (MAGE-C2) in tumor-free breast specimens, breast benign disease specimens and breast cancer specimens.Methods:Reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry were used to investigate the expressions of MAGE-C2 in 60 tumor-free breast specimens, 60 breast benign disease specimens and 60 breast cancer specimens. The correlation of MAGE-C2 expression with clinicopathological parameters and prognosis of breast cancer patients were analyzed. The expression of MAGE-C2 was also detected by RT-PCR in breast cancer cell MCF-7 and MDA-MB-231 treated with DNA methylase inhibitor 5-aza-2′-deoxycytidine (5-aza-CdR) and histone deacetylase inhibitor trichostatin A (TSA).Results:The positive expression rates of MAGE-C2 mRNA and protein were 61.7% (37/60) and 58.3% (35/60) in breast cancer specimens, respectively, while negative expressed in breast and begin disease specimens. MAGE-C2 protein expression was associated with tumor grade, histological type and blood vessel invasion of breast cancer patients ( P<0.05). The incidence of recurrence-free survival of patients with positive MAGE-C2 expression were lower than that of patients with negative MAGE-C2 expression ( P<0.05). Multivariate Cox regression analysis showed that the clinical stage ( P<0.01), lymph node metastasis ( P<0.05) and MAGE-C2 expression ( P<0.05) were the independent prognostic factors of breast cancer patients. The MAGE-C2 mRNA was not observed in the control and TSA treated breast cancer cells while upregulated in the 5-aza-CdR treated cells. Besides, 5-aza-CdR combined with TSA further enhanced MAGE-C2 mRNA level in breast cancer cells ( P<0.05). Conclusions:MAGE-C2 is one of the tumor-specific antigen and its expression is related with the poor prognosis of breast cancer patients. DNA methylation and histone acetylation may be an important regulation mechanism of MAGE-C2 gene expression.

Objective:To analyze the expression pattern, mechanism and clinical significance of melanoma-associated antigen-C2 (MAGE-C2) in tumor-free breast specimens, breast benign disease specimens and breast cancer specimens.Methods:Reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry were used to investigate the expressions of MAGE-C2 in 60 tumor-free breast specimens, 60 breast benign disease specimens and 60 breast cancer specimens. The correlation of MAGE-C2 expression with clinicopathological parameters and prognosis of breast cancer patients were analyzed. The expression of MAGE-C2 was also detected by RT-PCR in breast cancer cell MCF-7 and MDA-MB-231 treated with DNA methylase inhibitor 5-aza-2′-deoxycytidine (5-aza-CdR) and histone deacetylase inhibitor trichostatin A (TSA).Results:The positive expression rates of MAGE-C2 mRNA and protein were 61.7% (37/60) and 58.3% (35/60) in breast cancer specimens, respectively, while negative expressed in breast and begin disease specimens. MAGE-C2 protein expression was associated with tumor grade, histological type and blood vessel invasion of breast cancer patients ( P<0.05). The incidence of recurrence-free survival of patients with positive MAGE-C2 expression were lower than that of patients with negative MAGE-C2 expression ( P<0.05). Multivariate Cox regression analysis showed that the clinical stage ( P<0.01), lymph node metastasis ( P<0.05) and MAGE-C2 expression ( P<0.05) were the independent prognostic factors of breast cancer patients. The MAGE-C2 mRNA was not observed in the control and TSA treated breast cancer cells while upregulated in the 5-aza-CdR treated cells. Besides, 5-aza-CdR combined with TSA further enhanced MAGE-C2 mRNA level in breast cancer cells ( P<0.05). Conclusions:MAGE-C2 is one of the tumor-specific antigen and its expression is related with the poor prognosis of breast cancer patients. DNA methylation and histone acetylation may be an important regulation mechanism of MAGE-C2 gene expression.

Objective: To explore the roles and mechanisms of long non-coding RNA (lncRNA) small nucleolar RNA host gene 6 (SNHG6) in promoting invasion and metastasis of esophageal squamous carcinoma (ESCC). Methods: Real time quantitative polymerase chain reaction (qPCR) was used to detect the expression of SNHG6 in ESCC and matched para-carcinoma tissues. Reverse transcription-polymerase chain reaction (RT-PCR) was performed to detect the expression of SNHG6 in ESCC cell lines (TE1, Yes-2, Eca9706 and Kyse150). Then, TE1 cell line which harbored highest expression of SNHG6 was used in following experiments. siRNAs were used to knock down the expression of SNHG6. Clone formation, wound-healing and transwell assay were used to detect the abilities of proliferation, migration andinvasionofTE1cells,respectively.Westernblottingwasusedtodetecttheexpressions of MMP-2, MMP-9andZEB1 protein before and after knockdownofSNHG6inTE1cells.Results:SNHG6washighlyexpressedinESCC tissues, compared to para-carcinoma tissues (P<0.01). The expression of SNHG6 was significantly decreased after transfection of SNHG6siRNA (all P<0.01). The abilities of proliferation, migration and invasion of TE1 cells in si-SNHG6-1 and si-SNHG6-2 group were significantly lower than those in the control group (all P<0.01). The expressions of ZEB1, MMP-2and MMP-9 in si-SNHG6-1 and si-SNHG6-2 group were significantly lower than those in the control group (all P<0.05). Conclusion: SNHG6 is highly expressed in ESCC tissues and promotes the malignant biological behavior of ESCC cells. Its mechanism of promoting the occurrence and development of ESCC may be related to the upregulation of ZEB1 expression.

[Abstract] Objective: To investigate the expression of MAGE-C1 (melanoma-associated antigen-C1) in breast cancer tissues and its correlation with clinicopathological features and prognosis of breast cancer patients. Methods: Breast cancer tissues, normal breast tissues and benign breast lesion tissues (60 samples for each) were collected from the Fourth Hospital of Hebei Medical University during January 2008 and December 2008.The mRNA and protein expressions of MAGE-C1 in three types of breast tissues were detected by RT-PCR and immunohistochemistry, and their correlation with clinicopathological parameters and prognosis of breast cancer patients were also analyzed. DNA methylase inhibitor 5-aza-2'-deoxycytidine (5-Aza-CdR) and histone deacetylase inhibitor trichostatin A (TSA) were used to treat breast cancer MDA-MB-231 and MCF-7 cells, and RT-PCR was used to determine the changes in mRNA expression of MAGE-C1 after drug treatment. Results: The positive expression rate of MAGE-C1 mRNA and protein in breast cancer tissues were 43.3% (26/60) and 38.3% (23/60), respectively; and the mRNA and protein expressions of MAGE-C1 were all negative in normal breast tissues and benign breast lesion tissues. MAGE-C1 expression was positively associated with high tumor grade (χ2 =6.233, P<0.05). Recurrence-free survival (RFS) of patients with negative MAGE-C1 expression was significantly longer than those patients with positive MAGE-C1 expression (χ 2 =4.213, P<0.05). MAGE-C1 expression (HR=3.980, P<0.05) and clinical stage (HR=3.637, P<0.05) could be used as independent prognostic factors for breast cancer patients. 5-Aza-CdR and/or TSA treatment had no significant influence on MAGE-C1 gene expression (P>0.05). Conclusion: MAGE-C1 is a tumor-specific antigen and its expression is associated with poor prognosis of breast cancer patients.

Objective: To investigate the expression of miR-1269a in esophageal squamous cell carcinoma (ESCC) tissues and its effect on the malignant biological behaviors of ESCC KYSE30 cells, as well as to explore the underlying mechanism. Methods: Ninety specimens of ESCC tissues and adjacent para-cancerous tissues were obtained from patients underwent surgery in Fourth Hospital, Hebei Medical University. In addition, normal esophageal immortalized epithelial cells and esophageal cancer cell lines were also collected. The expression level of miR-1269a in above mentioned tissues and cell lines was examined by Real-time fluorescent quantitative PCR. After being transfected with miR-1269a mimics and inhibitors, the effects of miR-1269a on proliferation, migration, invasion and colony formation of KYSE30 cells were detected by MTS, Transwell and colony formation assay, respectively. The bioinformatics tool was used to predict the possible target genes of miR-1269a. Then the regulation effect of miR-1269a on target gene expression was validated by WB and Dual-luciferase reporter assay. After being transfected with SOX6 plasmid, the effects of SOX6 on the proliferation, migration, invasion and colony formation of KYSE30 cells were detected by MTS, Transwell and colony formation assay, respectively. At last, rescue assay was used to confirm the results. Results: The expression level of miR-1269a in ESCC tissues was significantly higher than that in adjacent para-cancerous tissues (P<0.05), and the expression level of miR-1269a in ESCC cell lines was significantly elevated compared with the normal epithelial cells (P<0.05 or P<0.01). The capacities of proliferation, invasion, migration and colony formation of KYSE30 cells in miR-1269a mimics transfection group were obviously higher than those in mimics NC group, while those abilities in miR-1269a inhibitor transfection group were significantly lower than those in inhibitor NC group (P<0.05 or P<0.01). Bioinformatics analysis showed that miR-1269a could combine with 3’UTR region at SOX6 gene; and after miR-1269a over-expression, the expression level of SOX6 and luciferase activity in KYSE30 cells were significantly reduced (P<0.05). Rescue assay showed that miR1269a over-expression could promote the proliferation, invasion and migration of KYSE 30 cells, while simultaneous transfection of SOX6 could partially reverse the promotion effect of miR-1269a mimics. Conclusion: The expression level of miR-1269a in ESCC tissues and cell lines is significantly increased, and it could enhance proliferation, migration, invasion and colony formation of KYSE30 cell line.And its mechanism may be related to the suppression of its target gene SOX6.

Objective: To investigate the expression of metastasis-associated protein 2 (MTA2) in human bladder cancer tissues and its effect on the malignant biological behaviors of bladder cancer T24 cells, as well as to explore the effect of MTA2 on the progression of bladder cancer. Methods: Sixty-two cases of human bladder cancer tissues and 28 cases of normal bladder tissues (from patients with cystitis, and pathologically confirmed as normal tissue) were collected at People’s Hospital of Hebei Province during December 2012 and December 2014. The expression of MTA2 in bladder cancer tissues and normal bladder tissues was detected by immunohistochemical staining, and the correlation between MTA2 expression and clinicopathological characteristics of patients was also analyzed. The bladder cancer T24 cell line stably expressing MTA2 was constructed. The effects of MTA2 on the proliferation, colony formation, migration and invasion of bladder cancer T24 cells were detected by MTS, clone formation, scratch healing and Transwell assay, respectively. Results: Immunohistochemical staining showed that MTA2 expression was significantly up-regulated in bladder cancer tissues as compared with normal bladder tissues (P<0.01). The high expression of MTA2 in bladder cancer tissues was not related to gender, age and tumor volume (P>0.05), but was associated with higher TNM stage, histological grade, and lymphatic infiltration and metastasis (all P<0.05). After over-expression of MTA2 in bladder cancer T24 cell line, the proliferation activity of the cells was significantly increased (P<0.05), and the colony formation, scratch healing, migration and invasion ability were significantly increased (all P<0.01). Conclusions: MTA2 is up-regulated in human bladder cancer tissues and can promote the proliferation, tumor formation, migration and invasion of T24 cells.