ABSTRACT
Objective:To find new biomarkers for the diagnosis and prognosis of sepsis through analyzing the differential expression protein in sepsis by proteomics and bioinformatics analysis and enzyme linked immunosorbent assay (ELISA).Methods:Patients with sepsis admitted to the emergency department of the Affiliated Hospital of Southwest Medical University from January to December 2019 were enrolled. And meanwhile, healthy volunteers who had normal physical examinations were included as the control group. Blood samples from two groups were collected. The samples were randomly selected for the protein concentration by data independent acquisition (DIA). Bioinformatics method was used in differentially expressed proteins by gene ontology (GO) pathway, enrichment analyses, groups meta-analysis and survival curves construction. ELISA method was used to verified marker screened. Then the data of transferrin receptor CD71 and the clinical data of procalcitonin (PCT), C-reactive protein (CRP) and blood lactic acid (Lac) were collected to construct receiver operator characteristic curve (ROC curve), and biomarker was screened for diagnostic and prognostic of sepsis.Results:The result of DIA showed that 71 differentially expressed proteins were screened out from sepsis group, 6 proteins were down-regulated and 65 proteins were up-regulated. Those differentially expressed proteins were enriched in the inflammatory response, response to stress, leukocyte migration in the GO pathway and enrichment analyses. The meta-analysis showed that the expression level of CD71 was higher in sepsis group than normal control group [standardized mean difference ( SMD) = -0.47, 95% confidence interval (95% CI) was -0.93 to 0.00, P < 0.01], the expression level of CD71 was higher in non-survivor group than survivor group ( SMD = -0.44, 95% CI was -0.70 to -0.18, P = 0.63). Survival curve showed that the expression of CD71 was inversely correlated to survival rates, the patients with a lower expression had higher survival rates ( P = 0.000 34); the ELISA showed that the level of CD71 was higher in sepsis group than normal control group (nmol/L: 156.83±84.71 vs. 87.99±47.89, P < 0.05), the level of CD71 was higher in non-survivor group than survivor group (nmol/L: 219.63±125.59 vs. 130.97±40.45, P < 0.05). The area under the ROC curve (AUC) of CD71 in diagnostic performance of sepsis was 0.790 (sensitivity was 65.1%, specificity was 90.0%), the AUC of CD71 in prognostic performance of sepsis was 0.744 (sensitivity was 57.1%, specificity was 94.1%); CD71 had a better prognostic performance than PCT (AUC = 0.547, sensitivity was 64.3%, specificity was 55.9%), CRP (AUC = 0.594, sensitivity was 64.3%, specificity was 61.8%), Lac (AUC = 0.540, sensitivity was 42.9%, specificity was 82.4%). Conclusion:CD71 had a great value of diagnostic and prognostic performance in sepsis, and it was expected to be a potential biomarker for sepsis.
ABSTRACT
Objective:To construct the prognostic prediction model and scoring tool by using severe trauma patients’ physiological indicators on admission, and to verify the clinical application effect and provide a reference for the early evaluation of severe trauma patients.Methods:This study was a retrospective study which adopted cluster sampling. Patients who met the inclusion and exclusion criteria in the emergency department of the First Affiliated Hospital of Soochow University from September 2019 to November 2020 were included. Patients were randomly assigned into the modeling group and the validation group in a ratio of 7:3 based on their outcome in the emergency department. Logistic regression analysis was performed to construct a prediction model, which was simplified as a scoring tool. The model was verified by using validation group and two months’ prospective validation. The efficiency of the simplified scoring tool was compared with that of the revised trauma score (RTS) and the injury severity score (ISS).Results:Totally 863 patients were included in this study, including 604 patients in the modeling group and 259 patients in the validation group. The model included systolic blood, SpO 2 and AVPU score. The AUC for predicting the death of severe trauma patients was 0.938. The AUC of the prediction model was 0.933, the best cut-off point was 5, the sensitivity was 86.7%, the specificity was 94.2%; the AUC of the validation was 0.885, the sensitivity was 83.3%, the specificity was 93.7%; and the AUC of prospective validation was 0.919, the sensitivity was 100%, and the specificity was 76.7%. The AUC of the RTS and ISS were 0.800 and 0.833, respectively. The AUC of RTS was lower than that of the simplified scoring tool constructed in this research. Conclusions:The prediction model and simplified scoring tool are better than RTS in predicting the outcome of emergency severe trauma patients, which are convenient for emergency medical staff to evaluate the severity of trauma patients.
ABSTRACT
Objective To assess the noninvasive index for diagnosing the degree of esophageal varices (EV) in patients with hepatitis B virus (HBV)-related cirrhosis. Methods The clinical data of 112 patients with HBV-related cirrhosis were studied retrospectively. The patients were divided into non-EV (NEV) group (30 cases) and EV group (82 cases) according to the results of gastroscopy. In EV group, there were mild varices in 21 cases (mild EV group), moderate varices in 47 cases (moderate EV group) and severe varices in 14 cases (severe EV group). The age, gender, platelets, glutamyl transpeptidase, prothrombin time, albumin, bilirubin, portal vein diameter, spleen vein diameter and thickness of spleen were compared, and the relationship was analyzed between each index and EV. Results There were no statistical differences in gender, age, albumin, bilirubin, prothrombin time and glutamyl transpeptidase between NEV group and EV group (P>0.05). The portal vein diameter, spleen vein diameter and thickness of spleen in EV group were significantly higher than those in NEV group:(14.1 ± 3.1) mm vs. (10.6 ± 2.3) mm, (8.9 ± 2.1) mm vs. (7.6 ± 1.6) mm and (4.8 ± 0.9) mm vs. (3.8 ± 1.0) mm, the platelets in EV group was significantly lower than that in NEV group:(86.8 ± 20.2) × 109/L vs. (163.5 ± 18.1) × 109/L, and there were statistical differences (P<0.01 or<0.05). The portal vein diameter, spleen vein diameter and thickness of spleen in moderate EV group and severe EV group were significantly higher than those that in NEV group and mild EV group: (13.5 ± 2.1) and (14.8 ± 3.6) mm vs. (10.6 ± 2.3) and (11.2 ± 3.1) mm, (8.3 ± 2.1) and (9.1 ± 1.1) mm vs. (7.6 ± 1.6) and (8.1 ± 1.9) mm, (4.7 ± 1.1) and (4.9 ± 0.9) mm vs. (3.8 ± 1.0) and (4.1 ± 1.2) mm, the platelet levels were significantly lower than those in NEV group and mild EV group: (72.8 ± 11.6) × 109/L and (63.8 ± 15.6) × 109/L vs. (163.5 ± 18.1) × 109/L and (100.2 ± 10.3) × 109/L, and there were statistical differences (P<0.05). The area under curve of response operating characteristic for predicting the presence of moderate and severe EV with portal vein diameter and platelets were 0.719 and 0.735, and the cut off value were 14 mm and 69 × 109/L. Conclusions The portal vein diameter and platelets can predict the presence of moderate and severe EV in patients with HBV-related cirrhosis.
ABSTRACT
10.3969/j.issn.2095-4344.2013.23.004
ABSTRACT
The transcriptional factor Oct-4 and Survivin are the key regulatory factors in cancer cell proliferation and mitosis. A dual cancer-specific shRNA adenovirus vector, Ad5-Dual-shRNA, targeting Oct-4 and Survivin genes was constructed by molecular cloning and recombination. After cells were infected with virus, hepatocellular carcinoma cell line EHBH-H1 was used for detecting the expression of Oct-4 and Survivin proteins by Western blotting. The viral cytotoxic effect on cancer cells was detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) reduction assay in vitro, and the inhibition effect on tumor xenografts was observed in nude mice. The results showed that the expression of Oct-4 and Survivin in cancer cell line EHBH-H1 could be silenced markedly by Ad5-Dual-shRNA. In MTT and animal experiments, Ad5-Dual-shRNA also represented much stronger anti-tumor effect on tumor growth than Ad5-Surv-shRNA and Ad5-Oct4-shRNA. From this research we can draw a conclusion that the cancer-specific adenovirus vector expressing dual-shRNA targeting Oct-4 and Survivin genes may provide us a more effective, specific and convenient gene therapy method.
Subject(s)
Animals , Humans , Mice , Adenoviridae , Genetics , Metabolism , Apoptosis , Genetics , Carcinoma, Hepatocellular , Pathology , Therapeutics , Cell Line, Tumor , Genetic Therapy , Genetic Vectors , Genetics , HEK293 Cells , Inhibitor of Apoptosis Proteins , Genetics , Liver Neoplasms , Pathology , Therapeutics , Mice, Nude , Octamer Transcription Factor-3 , Genetics , RNA, Small Interfering , Genetics , Recombinant Proteins , Genetics , TransfectionABSTRACT
Objective To compare the expression of mIFN-γ, replicative activities and anti-tumor activities of CNHK300-mIFN-γand Ad-mIFN-γin normal and gastric cancer cells. Methods The replicative activities of viruses in cells were measured by viral replication assay. CPE assay was used to detect the antitumor effect of viruses. The expression level of mIFN-γ in cancer cells was detected by ELISA. Results The infection of CNHK300-mIFN-γled to an obvious expression of mIFN-γin gastric cancer cells. The vector system CNHK300-mIFN-γpossessed more replicated potential than Ad-mIFN-γ, and could specifically kill most of BGC-823 cells at MOI value of 0.1, which was much better than that by the traditional adenoviral vector. Conclusion CNHK300-mIFN-γcan selectively replicate and effectively express mIFN-γ in tumor cells, and specifically kill gastric cancer cells, suggesting a splendid future as a new anticancer agent.
ABSTRACT
Objective To discuss the curative effect of unremitting pump infusion of microdose urokinase(100 000 u/24 h)into the cerebral venous sinus in treating thrombosis in cerebral venous sinus which had anatomical variation. Methods Mechanical disruption of the thrombus and unremitting pump infusion of microdose urokinase(100 000 u/24 h)into the cerebral venous sinus for 48-96 hours were employed in 9 patients with thrombosis in anatomically varied cerebral venous sinus.After the procedure the original disorder was actively treated and the anticoagulant therapy was continued for 6 months.A follow-up of 6-12 months(mean 10 months) was conducted. Results Recanalization of the previously occluded cerebral venous sinus was obtained in all 9 patients.The dose of urokinase was 100 000 u/24 h in 8 patients.For the remaining one patient the dose of urokinase was 100 000 u/24 h in the first 48 hours,then the dose Was increased to 250 000u/24 h. Excellent result was obtained in all patients.Conclusion Unremitting pump infusion of microdose urokinase into the cerebral venous sinus can effectively treat the thrombosis in anatomically varied cerebral venous sinus.
ABSTRACT
BACKGROUND:It has been found that central nervous system is involved in Guillain-Barre syndrome and Miller-Fisher syndrome, and the involved sites include optic nerve, brain stem and cerebellum. Abnormal signal of MRI can be observed in the brainstem and spinocerebellar tract of patients with Miller-Fisher syndrome. To establish an animal model of encephalitis after infection of Campylobacter jejuni, and investigate the mechanism of formation by means of imaging, immunology and pathology.OBJECTIVE: To construct an animal model of lesion of central nervous system after infection of Campylobacter jejuni Penner 4.DESIGN: A randomized grouping designed, controlled animal experiment.SETTING: Department of Imaging and Department of Neurology, Second Hospital of Hebei Medical University.MATERIALS: The experiment was carried out in the Laboratory of Molecular Imaging, the Second Hospital of Hebei Medical University between August and December 2003. Fifteen healthy flap-eared rabbits were randomly divided into experimental group (n=10) and control group (n=5).METHODS: In the experimental group, Campylobacter jejuni inactivated bacteria liquor was completely emulsified with complete Freund adjuvant (CFA) of the same volume in week 1, and then the rabbits were immunized with subcutaneous injection at multiple points of bilateral axilla, bilateral groins and side of back spine, 1 mL for each site, and 5 mL for each rabbit; The rabbits were further immunized with intraperitoneal injection of simple Campylobacterjejuni inactivated bacteria liquor in the following every two weeks, 5 mL for each time in each rabbit for 5 times. In the control group, the Campylobacter jejuni inactivated bacteria liquor was replaced by saline of the same volume, the injected method and time were all the same as those in the experimental group. Evaluative methods: ①Symptoms and physical signs: their mental status, conditions of diet, urine and excrement, and activities of limbs were observed; ② Serological examination: the contents of anti-Campylobacterjejuni antibody, anti-IgG GM1 antibody and myelin basic protein (MBP) were detected with enzyme-linked immunoadsorbent assay (ELISA); ③ MRI examination was applied to the randomly selected rabbits before every immunization with Toshiba 1.5 T MRI instrument. The scanning sequence included spin-echo T1-weighted image with the scanning parameter of 500/15 ms (TR/TE); rapid spin-echo T2-weighted image, 4 000/108 ms (TR/TE); fluid attented inversion recovery (Flair) sequence, the parameter was 10 000/120 ms (TR/TE), inversion angle was 90°. The thickness of scanning layer was 4.0 mm, and the layer space was 0.8 mm. ④ Histological examination: At 4 weeks after the first immunization, the attacked animals were induced to death by cardiac perfusion, and the skull was opened immediately to remove optic nerve, part white matter, hippocampus, brainstem, cerebellum and spinal cords of neck, chest and waist, which were fixed with formaldehyde solution (40 g/L),and hematoxylin-eosin (HE) staining, fast blue staining and MBP immunohistochemical staining were performed respectively. At 10 weeks after immunization, 5 randomly selected rabbits in the experimental group and the 5 rabbits in the control group were treated with the same methods to obtain the histological samples.MAIN OUTCOME MEASURES: The symptoms and physical signs,contents of anti-Campylobacterjejuni antibody, anti-IgG GM1 antibody and MBP, imaging observation and histological examination were mainly observed.RESULTS: Fifteen animals were enrolled, 14 were involved in the analysis of results, 1 rabbit in the experimental group died at 4 weeks after immunization. ① Mental symptoms and disorder of limb's activity occurred in 1 rabbit in the experimental group at 2 weeks after immunization. ② In the experimental group, titre of anti-Campylobacterjejuni-IgG antibody in serum reach the peak at 2-4 weeks. From week 2, the serum A value was significantly higher in the experimental group than in the control group (1.923±0.403, 0.973±0.633, P ＜ 0.05). The IgG type GM1 (A value) was obviously elevated at week 8, but insignificantly different from that in the control group (0.115±0.042, 0.097±0.039, P ＞ 0.05). The MBP content (Avalue) in serum was significantly elevated at the 8th week (0.134±0.041).③ The imaging examination showed that abnormal MRI signal of different degree occurred at 2-4 weeks after immunization in the experimental group. ④ The histological changes showed that there was swelling of myelin sheath at the sites of brainstem, medulla oblongata, cervical spinal cord, thoracic spinal cord and lumbar spinal cord in the experimental group, no inflammatory cell infiltration and deletion of myelin sheath were observed. No obvious changes at the above site were observed in the contro1 group.CONCLUSION: Campylobacterjejuni Penner 4 can induce lesion of central nervous system.
ABSTRACT
AIM To investigate whether dipfluzine (Dip) possesses antiarrhythmic effect on experimental arrhythmias and effect on cytosolic calcium in ventricular myocytes of guinea-pig. METHODS Experimental arrhythmias were induced by strophanthin G infusion through jugular vein in guinea-pigs and by myocardial ischemia-reperfusion (I-R) in rats respectively. Cytosolic calcium concentration ([Ca2+]i) of isolated guinea-pig ventricular myocytes was examined with laser confocal scanning microscope. RESULTSIn guinea-pigs pretreatment with Dip 20 mg·kg-1 increased the dosages of strophanthin G required to induce ventricular premature contraction (VP), ventricular tachycardia (VT), ventricular fibrillation (VF) and cardiac arrest (CA), pretreatment with Dip 10 mg·kg-1 increased the dosages of strophanthin G required to induce VP. In the I-R-induced arrhythmic model of rats, Dip 20 mg·kg-1 decreased the number of rats exhibiting VT, VF and CA, and the number of rats exhibiting VF and CA was decreased by Dip 10 mg·kg-1. Both Dip and verapamil (Ver) decreased [Ca2+]i of the ventricular myocytes in normal Tyrode′s solution. The Ca2+ overload evoked by high extracellular Ca2+ levels was inhibited by Dip and Ver, and the prophylactic effect of Dip was less than that of Ver, while the curative effect of Dip was more obvious than that of Ver. CONCLUSION Dip has antiarrhythmic effect, which is likely related to the modulation on the intracellular calcium homeostasis.
ABSTRACT
Objective:To construct a replication-deficient adenovirus carrying p16 gene and to investigate its anti-tumor activity on human gastric cancer xenografts in nude mice.Methods:p16 cDNA was amplified by PCR and was inserted into the plasmid pSuCMV,the latter was then used to recombine the replication-deficient adenovirus AdCMV-p16 in 293 cells.Human SGC-7901 gastric cancer xenograft models were established in nude mice and were divided into 3 groups:AdCMV-p16,Ad-LacZ,and control groups.Mice in AdCMV-p16 group received intratumoral injections of 2?108 pfu/100 l AdCMV-p16(injected every other day for 5 times).Mice in control group received the same volume of virus preserving solution.The tumor volumes were measured at predefined time points.The anti-tumor effect of AdCMV-p16 was observed by p16 immunochemical study and TUNEL detection of cell apoptosis.Results:The replication-deficient adenovirus expressing p16 gene evidently inhibited the growth of human gastric cancer xenografts in nude mice(P0.05),only with a inhibition rate of 4.26%.The pathological examination showed that apoptoses were the main changes in AdCMV-p16 group,and p16 gene was found in the cancer cells.Conclusion:The replication-deficient adenovirus harboring p16 gene can recover the expression of p16 in gastric cancer cells and subsequently inhibit the growth of human gastric cancer.