Construction and anti-tumor efficacy of a peptide vaccine that targets calcium-binding protein S100A9 / 药学学报

Calcium-binding protein S100A9 is closely related to inflammation and tumor invasion, and is one of the specific markers of myeloid-derived suppressor cells (MDSC). In this study, a recombinant polypeptide vaccine CTB-S100A9 targeting mouse calcium-binding protein S100A9 was constructed by fusion cholera toxin B subunit (CTB) with S100A9 gene. The CTB-S100A9 fusion protein was expressed in E coli. and purified by Ni+ affinity chromatography. Vaccinate the purified recombinant CTB-S100A9 protein supplemented with aluminum hydroxide adjuvant can break the autoimmune tolerance and produce high titer of S100A9 antibody in mice. Moreover, the S100A9 antibody produced by CTB-S100A9 vaccination is more specific and does not cross-react with S100A8. In the mouse 4T1 breast cancer model, CTB-S100A9 vaccination not only has significant tumor prevention effects, but also has significant tumor therapeutic effects. In addition, CTB-S100A9 significantly inhibited lung metastasis in 4T1 mice breast cancer model. Further analysis by flow cytometry showed that CTB-S100A9 vaccination can significantly reduce the tumor induced Treg cells and granulocyte-derived MDSC in 4T1 mice model, and reverse the tumor immunosuppressive environment, thereby promote the anti-tumor efficacy. The animal experiments in this study were carried out under the animal care guidelines approved by the Animal Ethics Committee of the Affiliated Hospital of Integrated Traditional Chinese and Western Medicine, Nanjing University of Chinese Medicine. This study shows that CTB-S100A9 is a good recombinant vaccine that targets the tumor immune-suppression environment and has great potential for the future clinical application.

Analysis of clinical phenotypes and CYP21A2 gene variants in 18 patients with 21-hydroxylase deficiency / 中华医学遗传学杂志

OBJECTIVE@#To explore the genotype-phenotype correlation among 18 patients with 21-hydroxylase deficiency (21-OHD).@*METHODS@#PCR-Sanger sequencing was used to analyze the 10 exons and flanking regions of the CYP21A2 gene among the 18 patients and 20 healthy controls.@*RESULTS@#Seventeen patients had variants of the CYP21A2 gene. Eight patients (44.4%, 8/18) carried homozygous variants including p.Ile 173Asn (62.5%, 5/8), p.Pro31Leu (25.0%, 2/8), and IVS2-13A/C>G (12.5%, 1/8), respectively. Six patients (33.3%, 6/18) carried compound heterozygous variant, among which IVS2-13 A>G+p.Ile 173Asn were most common (50.0%). 94.4% (34/36) of the variant were pathogenic, with the most common variants being p.Ile173Asn (41.7%), IVS2-13A/C>G (19.4%), and p.Ile173Asn (7.5%). No variant was identified among the 20 healthy controls.@*CONCLUSION@#The majority of 21-OHD patients carried CYP21A2 gene variants in homozygous or compound heterozygous forms, among which the p.Ile173Asn was the most common one. There is a strong correlation between the genotypes and clinical phenotypes.

Preparation and Application of Porous Layer Open Tubular Capillary Columns with Narrow Bore in Liquid Chromatography / 分析化学

In recent years, micro/nano-scaled separation technique has attracted increasing attention due to its inherent advantages. The porous layer open tubular ( PLOT) capillary column is an important microcolumn form. Comparied to wide-bore PLOT capillary columns ( inner diameter >25 μm ) , the PLOT capillary columns with a narrow inner diameter yield higher separation efficiency and lower reagent consumption. In this paper, the preparation methods for porous layer open tubular capillary columns with narrow inner diameter, less than or equal to 25 μm, are reviewed. Detection techniques combined with mass spectrometry and their applications in liquid chromatography are also disussed.

NF-κB participates in the regulation of apoptosis / 上海交通大学学报（医学版）

The abnormal changes in the apoptosis activities are relevant to a number of diseases including the tumor genesis and development. Since the gene structure of NF-κB being cloned three decades ago, the investigation of the transcriptional factor has been improved over time. A series of target genes are revealed to be modulated, which participate in physiological and pathological activities, including inflammation, proliferation, differentiation,senescence, and apoptosis. This review is focused on the signaling network of cell apoptosis regulated by NF-κB, the molecular involved, as well as the environmental variants associated. Furthermore, as NF-κB is unusually up-regulated in diseases such as cancer, searching for its specific antagonists may prevent the occurrence of those diseases.

NF-κB participates in the regulation of apoptosis / 上海交通大学学报（医学版）

The abnormal changes in the apoptosis activities are relevant to a number of diseases including the tumor genesis and development. Since the gene structure of NF-κB being cloned three decades ago, the investigation of the transcriptional factor has been improved over time. A series of target genes are revealed to be modulated, which participate in physiological and pathological activities, including inflammation, proliferation, differentiation,senescence, and apoptosis. This review is focused on the signaling network of cell apoptosis regulated by NF-κB, the molecular involved, as well as the environmental variants associated. Furthermore, as NF-κB is unusually up-regulated in diseases such as cancer, searching for its specific antagonists may prevent the occurrence of those diseases.

Gastro-protecting effect of gefarnate on chronic erosive gastritis with dyspeptic symptoms / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The role of gastro-protecting agents on symptomatic chronic gastritis is unclear. This multicenter, open, randomized trial was designed to compare the comprehensive effects of gefarnate with sucralfate on erosive gastritis with dyspeptic symptoms.</p><p><b>METHODS</b>Totally 253 dyspepsia patients confirmed with erosive gastritis were enrolled from six centers in China. They randomly received either daily 300 mg gefarnate or 3 g sucralfate for six weeks. The primary endpoint was the effective rate of both treatments on endoscopic erosion at week six.</p><p><b>RESULTS</b>Gefarnate showed an effective rate of 72% and 67% on endoscopic score and dyspeptic symptom release, which is statistically higher than sucralfate (40.1% and 39.3%, P < 0.001, intension-to-treat). For histological improvement, gefarnate showed both effective in decreasing mucosal chronic inflammation (57.7% vs. 24.8%, P < 0.001, intension-to-treat) and active inflammation (36.4% vs. 23.1%, P < 0.05, intension-to-treat) than the control. A significant increase of prostaglandins and decrease of myeloperoxidase in mucosa were observed in gefarnate group. Severity of erosion is non-relevant to symptoms but Helicobacter pylori (H. pylori) status does affect the outcome of therapy.</p><p><b>CONCLUSIONS</b>Gefarnate demonstrates an effective outcome on the mucosal inflammation in patients with chronic erosive gastritis. Endoscopic and inflammation score should be the major indexes used in gastritis-related trials.</p>

Endoscopic retrograde cholangiopancreatography-guided brush cytology diagnosis of pancreatobiliary tumors / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the cytologic features of pancreatobiliary tumors in endoscopic retrograde cholangiopancreatography (ERCP)-guided brushing preparations and to evaluate the usefulness of cytology in the diagnosis of pancreatobiliary malignancy.</p><p><b>METHODS</b>A retrospective analysis of 212 cases of ERCP-guided brush cytology smears performed during the period from January, 2004 to December, 2006. The cytologic diagnosis was confirmed either by the histologic diagnosis or the strict clinical criteria.</p><p><b>RESULTS</b>Two of the cases studied were unsatisfactory for diagnosis, with no epithelial cells identified. One hundred and thirty-seven smears were diagnosed as "negative", 45 of which subsequently confirmed to be malignant (negative predictive value = 60.2%). Six of the 11 cases with "low-grade atypia" were proven to be malignant (positive predictive value = 54.5%), as compared to 19 of 23 cases of "high-grade atypia" (positive predictive value = 86.4%). All of the 41 cases with cytologic diagnosis of "malignancy" were confirmed to be malignant (positive predictive value = 100%). The cytologic features of malignancy in ERCP-guided brushing preparations included overlapping nuclei, anisonucleosis, coarse chromatin pattern, poor cellular cohesion, tumor diathesis, prominent nucleoli and atypical mitotic figures.</p><p><b>CONCLUSIONS</b>The accuracy of ERCP-guided brush cytology relies on good specimen preparation and application of morphologic criteria. Grading of cytologic atypia is of clinical significance. A "negative" or "low-grade atypia" cytologic diagnosis requires further diagnostic workup to rule out the possibility of underlying malignancy, while a "high-grade atypia" or "malignant" diagnosis is relatively specific in guiding subsequent management of suspected pancreatobiliary malignancy.</p>

Aberrant methylation of CpG island in 5′promoter region of p16 gene and its role in diagnosis of pancreatic cancer / 第二军医大学学报

Objective:To investigate the aberrant methylation of CpG island in 5′promoter region of p16 gene in the pancreatic juice and its value in diagnosis of patients with pancreatic cancer.Methods:Pure pancreatic juice(PPJ)was collected from the pancreatic duct by a nasopancreatic tube put under endoscopic retrograde cholangiopancreatography(ERCP). Cytological examination was performed by H-E staining in pure pancreatic juice.Aberrant p16 methylation was detected using the methylation specific PCR(MSP)in the PPJ.Results:The sensitivity,specificity,positive predictive value,negative predictive value and accuracy cytological examination in diagnosis of pancreatic cancer were 40%,100%,100%,45.4% and 60.0%,respectively.The DNA sequences were successfully extracted from the PPJ of 30 patients with pancreatic diseases and were subjected to MSP.Seven(35%)of the 20 cases with pancreatic cancer showed aberrant methylation of p16 gene.No aberrant methylation was detected in the pancreatic juice samples of patients with chronic pancreatitis and mucinous cystoadenocarcinoma of pancreas.When cytological examination combined with p16 methylation detection,the sensitivity, specificity,positive predictive value,negative predictive value and accuracy for diagnosis of pancreatic cancer were 55%,100%, 100%,52.6% and 70%,respectively.Conclusion:Pancreatic juice collected by nasopancreatic drainage during ERCP can be used for molecular analysis.Detection of aberrant methylation of p16 gene in pancreatic juice combined with cytological examination is a better method for diagnosis of pancreatic cancer.

Role of extracellular signal-regulated kinase 1/2 in pathogenesis of stress-induced gastric ulcer / 第二军医大学学报

Objective:To investigate the role of extracellular signal-regulated kinase 1/2(ERK1/2)signal pathway in the pathogenesis of stress-induced gastric ulcer.Methods:Animal model of stress-induced gastric ulcer was established in rats with water-immersion restraint(WIR)stress.The mucosal activation of ERK1/2 was observed before and 5,15 and 30 min,and 1, 2 and 3.5 h after WIR stress.Some animals were also treated with an intravenous injection of PD98059(1 mg/kg),a specific ERK1/2 inhibitor,1 h prior to WIR stress.Expression of total ERK1/2 and caspase-3 were detected by Western blot analysis; ERK1/2 activity was measured by kinase activity assay using myelin basic protein as a specific substrate.DNA-binding activities of the transcription factors activator protein-1(AP-1)and nuclear factor-?B(NF-?B)were determined by electrophoretic mobility shift assays(EMSA).Mucosal TNF-?and IL-1?mRNA expression was analyzed by Northern blot analysis.The degrees of the gastric mucosal lesions were expressed as ulcer index(UI)and pathological evaluation.Apoptosis in the gastric mucosa was examined by an in situ TdT-mediated dUTP nick-end labeling(TUNEL)method.Results:Activated ERK1/2 was very weakly expressed in the gastric mucosa of normal rats.ERK1/2 was rapidly activated in the gastric mucosa of rats 15 min after WIR stress and the activity reached the maximal after 3.5 h.Pretreatment with PD98059 significantly inhibited ERK1/2 activation,decreased AP-1 and NF-?B activities and TNF-?and IL-1?mRNA expression,and obviously relieved gastric mucosal lesions,accompanied by caspase-3 activation and increased apoptosis.Conclusion:The present results indicate that ERK1/2 activation plays an important role in the development of stress-induced gastric ulcer.