In order to assess the cytotoxin effect of Campylobacter jejuni (C.jejuni), the whole cell lysates from 9 strains of Guillan-Barre Syndrome (GBS)- associated C.jejuni and 4 strains of diarrhea-associated C. jejuni were co-cultivated with HeLa cells at the concentration of 0.001-5.00 μg/mL in vitro. The morphologic change of HeLa cells was observed under Olympus BX51 microscope after treatment with different concentration of bacteria lysate in the following 4 days. The morphological changes including swelling, irregularity and lysis of the affected cells over 50% was selected as the cut off for positive change and C.jejuni 81-176 and Helicobacter J99 strains were chosen as the positive and negative control. It was found that the minimum concentration to induce the positive changes in 10 strains(8 GBS associated and 2 strains from diarrhea patients)was 0.1μg/mL and 1 strain with the positive change at the minimum concentration of 1 μg/mL. There was only one GBS-associated strain causing no morphologic change on HeLa cells at the concentration of 5 μg/mL. It was evident that the cytotoxic effect of C.jejuni strains on HeLa cell was strain-specific, and there was no significant difference in the cytotoxic effect on HeLa cell between GBS-associated and diarrhea associated C.jejuni strains.

Background:Helicobacter pylori(Hp)is an important pathogen for peptic ulcer and gastric cancer,and is reportedly associated with a variety of extragastrointestinal diseases. However,there is no body fluid detection technique for Hp infection in clinical practice. Aims:To identify Hp infection-associated differentially expressed proteins in urine with relative molecular mass more than 10 kDa and provide potential biomarkers for diagnosis of Hp infection through body fluid detection. Methods:Midstream urine was collected from volunteers in the morning,and 13 C-urea breath test was performed to determine Hp infection. Each of 15 Hp-negative and 15 Hp-positive urine samples were mixed respectively for protein extraction. Spectra data were acquired by high performance liquid chromatogram-mass spectrometry,and label-free technology was used for relative quantitative analysis. The other 26 urine samples(15 Hp-negative and 11 Hp-positive) were used for validation by full scan. IPA software was employed for bioinformatics analysis. Results:A total of 475 urinary proteins were detected by label-free quantitative analysis and 42 differentially expressed proteins were identified. Finally,11 significantly up-regulated differentially expressed proteins were confirmed by external scanning validation. Bioinformatics analysis revealed the molecular functions,biological pathways,and related diseases of these differentially expressed proteins. Conclusions:These 11 differentially expressed proteins more than 10 kDa identified in urine might be potential biomarkers for diagnosis of Hp infection and provide molecular evidence for the correlation of Hp infection with extragastrointestinal diseases.

Objective To research the differential expression of trace phosphorylated proteins in human gastric adenocarcinoma epithelial (AGS) cells infected by Helicobacter pylori. Methods H. pylori 26695 strain infected AGS cells 4 h and AGS cells was cultivated for 4 h as a comparison. The proteins of AGS and comparison AGS cells were extracted. Their phosphorylated proteins were enriched by metal ion af-finity adsorption enrichment techniques. After desalinated and purified the phosphorylated proteins samples were separated by 2-dimensional polyacrylamide gel electrophoresis (2-DE) technique. Computer assisted image analysis was used to analyze the differential proteomic expression. The significantly differentially ex-pressed proteins were unambiguously assigned identities by matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF/TOF). Results Fifteen kinds of proteins were down-regulated, 4 kinds of new proteins were observed, 1 kind of proteins were up-regulated, 1 kind of proteins unexpression. The 21 proteins that were significantly differentially expressed , including cellular calcium ion homeostasis, transcription, interpretation, protein folding and transport, ribosomal assembly, centrosome replication, chromosome stability, cellular structure, cellular proliferation and apoptosis. Conclusion H. priori can cause a wide range change to human gastric adenocarcinoma epithelial cell protein pheshorylation. This change character has great significance to further comprehensive understanding of the pathogenesis of H. pylori.

Objective:To explore the related prognostic factors in patients with aortic dissection (AD) after surgery or interventional therapy, and to construct a multi-factor Cox regression analysis nomogram prediction model.Methods:The clinical data of patients diagnosed with AD in the emergency department of Affiliated Hospital of Jining Medical University of Shandong Province from January 2019 to December 2021 were collected to analyze retrospectively. The survival time was followed up, the survival curve was drawn by Kaplan-Meier method, the difference of survival rate among different Stanford types was tested by Log-Rank test, and Cox regression univariate and multivariate survival analysis was used to screen the prognostic factors. According to the results of multivariate analysis, the death risk nomogram model after AD was established, the differentiation of the model was evaluated by C-index, the accuracy of the model was evaluated by calibration curve, and the clinical benefit of the model was evaluated by decision curve analysis (DCA).Results:A total of 256 patients with AD were included, the mortality rate was 4.45% (37/256), and the overall survival rates at 3 months, 6 months and 12 months after operation were 90.23%, 87.50% and 85.55%, respectively. There was no significant difference in postoperative survival rate among AD patients with different Stanford types (χ 2=2.30, P=0.13). Cox multivariate regression analysis showed that history of hypertension ( HR=3.791, 95% CI 1.150-12.501, P=0.029), number of branch vessels involved ( HR=1.210, 95% CI 1.029-1.422, P=0.021), preoperative left ventricular ejection fraction (LVEF) ( HR=0.936, 95% CI 0.880-0.996, P=0.038) and perioperative complications ( HR=4.024, 95% CI 1.839-8.807, P<0.001) were independent prognostic factors in patients with AD. Furthermore, the predictive nomogram model of 3-month, 6-month and 12-month survival rate in patients with AD was constructed, and the C-index was 0.778. The calibration curve indicated that the accuracy of the model was good, and the clinical benefit of the DCA model was good. Conclusions:Hypertension history, the number of branch vessels involved, preoperative LVEF and perioperative complications are independent risk factors of postoperative death in patients with AD. The nomogram prediction model based on the above factors can be used to evaluate the postoperative survival of patients with AD.

Objective:To investigate the effects of survival status in Stanford type A aortic dissection (TAAD) patients over 70 years old by different treatment methods.Methods:The clinical data of 151 TAAD patients over 70 years old from January 2012 to January 2022 in the Affiliated Hospital of Jining Medical University were retrospectively analyzed. Among them, 60 patients were treated with surgery (surgical treatment group), and 91 patients with conservative method (conservative treatment group). The duration of hospitalization and complications (including pericardial effusion, acute myocardial infarction, pericardial tamponade, shock, stroke, mesenteric ischemia and acute renal failure) were recorded. The patients were followed up within 30 d after the onset of illness. The survival status was recorded. The multivariate Cox regression analysis was used to analyze the independent risk factors of death in TAAD patients over 70 years old.Results:The duration of hospitalization in surgical treatment group was significantly longer than that in conservative treatment group: 14.00 (7.00, 19.75) d vs. 5.00 (2.00, 10.00) d. The incidences of pericardial tamponade, shock and acute renal failure: 1.7% (1/60) vs. 13.2% (12/91), 8.3% (5/60) vs. 24.2% (22/91) and 0 vs. 9.9% (9/91), with statistical differences ( P<0.01 or <0.05), but no statistical differences in the incidences of pericardial effusion, acute myocardial infarction, stroke, mesenteric ischemia between two groups ( P>0.05). Patient follow-up for 30 d, the mortality rate in surgical treatment group was significantly lower than that in conservative treatment group: 15.0% (9/60) vs. 46.2% (42/91), with statistical difference ( χ2 = 15.69, P<0.01). The multivariate Cox regression analysis result showed that conservative treatment, female, increased aortic root diameter and concomitant stroke were the independent risk factors of death in TAAD patients over 70 years old ( RR = 2.311, 2.135, 1.051 and 3.737; 95% CI 1.056 to 5.057, 1.083 to 4.212, 1.004 to 1.100 and 1.393 to 10.026; P<0.05 or <0.01). Conclusions:The early surgical treatment is recommended in TAAD patients over 70 years old with surgical indications.

Objective:To explore the application value of evaluation and management of patients with acute chest pain in China (EMPACT) score in risk stratification for patients with acute chest pain.Methods:According to the methods of prospective cohort study, 548 patients with chest pain in the Affiliated Hospital of Jining Medical University from February to April 2021 were selected. The risk stratification was performed according to EMPACT score. The primary endpoint was the major adverse events (MAE) within 30 d, including death from all causes, acute myocardial infarction (AMI), emergency revascularization, cardiac arrest, cardiogenic shock and other life-threatening situations that need urgent attention. The receiver operating characteristic (ROC) curve was used to evaluate the predictive value of EMPACT score for MAE.Results:Among the 548 patients, 75 cases had MAE within 30 d (MAE group), and the incidence of MAE was 13.7%; 473 cases did not occur MAE (non-MAE group). The EMPACT score in MAE group was significantly higher than that in non-MAE group: 8 (12, 18) scores vs. 5 (2, 8) scores, and there was statistical difference ( Z = 8.94, P<0.01). Spearman correlation analysis result showed that EMPACT score was positively correlated with MAE ( r = 0.38, P<0.01). ROC curve analysis result showed that the area under the curve of EMPACT score in prediction within 30 d MAE was 0.820 (95% CI 0.770 to 0.871), the cut-off value was 9.5 scores (since all the scoring systems were integers, the cut-off value was 10 scores), the sensitivity was 88.6%, and the specificity was 60.0%. Conclusions:The EMPACT score has a good risk stratification capability to achieve safe and effective triage of acute chest pain.

Objective:To assess the application of three risk stratification scoring systems in evaluation and management of patients with acute chest pain.Methods:Patients with chest pain who visited the emergency department of Affiliated Hospital of Jining Medical University from February 2021 to April 2021 were recruited. The risk stratification evaluation was performed with EMPACT, HEART-Pathway and EDACS-ADP scoring systems. The primary endpoint was the major adverse events (MAE) within 30 days.The application values of three scales in identifying high-risk chest pain were evaluated.Results:A total of 628 patients with acute chest pain were enrolled, and 92 of them(14.95%) had MAE within 30 days. The scores of three scales were all positively correlated with MAE occurrence, while the EMPACT score had the highest correlation( r=0.41, P<0.001). ROC curve analysis showed that the area under the curve (AUC) of EMPACT score, HEART score and EDACS for predicting MAE within 30 days was 0.834(95% CI:0.790-0.878), 0.763(95% CI:0.710-0.817) and 0.635(95% CI:0.578-0.691), respectively. When the cut-off value was 9.5, the Yorden index of EMPACT score was the highest (0.561). Since all the scoring systems used integers, the EMPACT score of 10 was the threshold to distinguish low-risk chest pain from high-risk chest pain. The sensitivity of EMPACT, HEART-Pathway and EDACS-ADP scores in identifying high-risk chest pain patients was 0.707, 0.576 and 0.783, and the specificity of them was 0.854, 0.882 and 0.509, respectively. Conclusion:The EMPACT score has a good risk stratification ability, and it can be used for identifying patients with acute chest pain.

Objective:To investigate the survival status of patients with acute aortic dissection (AD) treated by different methods, and evaluate the factors affecting the survival of AD patients.Methods:According to the retrospective research method, the patients diagnosed with AD in the Affiliated Hospital of Jining Medical University from January 1, 2019 to December 31, 2020 were collected. The treatment data and follow-up of patients were counted, the survival of patients was analyzed by Kaplan-Meier survival curve, and the factors affecting the survival of patients were analyzed by Cox regression model.Results:A total of 251 patients were included in this study, including 169 patients in the surgical treatment group and 82 patients in the conservative treatment group. The in-hospital fatality rate in the surgical treatment group and the conservative treatment group were 8.88%(15/169) and 43.90%(36/82), respectively, and the difference were statistically significant ( P<0.05). The survival time of surgical treatment group and conservative treatment group was (328.08±8.17)d and (194.43±19.80)d, respectively, and the difference was statistically significant ( P<0.05). Surgery ( RR=5.424, 95% CI: 2.821-10.428, P<0.05), acute myocardial infarction ( RR=0.448, 95% CI: 0.221-0.906, P<0.05), and shock ( RR=0.266, 95% CI: 0.102-0.693, P<0.05) and stroke ( RR=0.277, 95% CI: 0.127-0.604, P<0.05) were the factors affecting 1-year survival in AD patients. Conclusions:Active surgical treatment is recommended for AD patients with surgical indication as soon as possible.

Background: Blackened intestines in slaughtered pigs have been commonly observed in China in recent years. However, no cause has been reported. Objectives: We attempted to determine whether the blackening of the pig intestine was related to an excess of copper (Cu) in their feed. Methods: In this study, we observed and collected porcine intestines in small- and large-scale pig slaughterhouses in Shandong province from May to October 2018. Twelve types of metal ions were detected in the black intestinal samples. Results: The Cu level in the intestine samples was mostly higher than the Chinese national limit for food. Further study showed that Cu supplementation in most commercial porcine feed also exceeded the national standard. An animal model (mouse) that could mimic the intestinal blackening in pigs was established. Compared to control mice, Cu accumulated in the liver and intestines of mice fed an excessive Cu level, confirming the excessive Cu in the feed may be considered the major cause of blackened porcine intestines. Microscopic examination revealed that black intestines had many particles containing Cu in the lamina propria of the intestinal mucosa, and the intestinal mucosal epithelial cells showed degeneration and necrosis. Conclusions In conclusion, overuse of Cu in animal feed can lead to animal poisoning and Cu accumulation in animal products. Such overuse not only harms the health of livestock but can also affect public health.

Background: Pseudorabies, also known as Aujeszky's disease, is caused by the pseudorabies virus (PRV) and has been recognized as a critical disease affecting the pig industry and a wide range of animals around the world, resulting in great economic losses each year. Shandong province, one of the most vital food animal-breeding regions in China, has a very dense pig population, within which pseudorabies infections were detected in recent years. The data, however, on PRV epidemiology and coinfection rates of PRV with other major swine diseases is sparse. Objectives: This study aimed to investigate the PRV epidemiology in Shandong and analyze the current control measures. Methods: In this study, a total number of 16,457 serum samples and 1,638 tissue samples, which were collected from 362 intensive pig farms (≥ 300 sows/farm) covered all cities in Shandong, were tested by performing enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR). Results: Overall, 52.7% and 91.5% of the serum samples were positive for PRV-gE and -gB, respectively, based on ELISA results. In addition, 15.7% of the tissue samples were PCR positive for PRV. The coinfection rates of PRV with porcine circovirus type 2 (PCV2), porcine reproductive and respiratory syndrome virus, and classical swine fever virus were measured; coinfection with PCV2 was 35.0%, higher than those of the other two viruses. Macroscopic and microscopic lesions were observed in various tissues during histopathological examination. Conclusions The results demonstrate the PRV prevalence and its coinfection rates in Shandong province and indicate that pseudorabies is endemic in pig farms in this region. This study provides epidemiological data that can be useful in the prevention and control of pseudorabies in Shandong, China.