To study the regulating effect of Astragalus Polysaccharides ( APS) to the mice infected by Brucella suis S2.Methods:120 BALB/c mice were randomly divided into 4 groups:experimental mice were injected APS 1 ml ( 0.4,1.2,3 mg/ml) via peritoneal cavity respectively once a day and the control group was injected with the same volume of saline for 3 days,then infected with Brucella suis S2 1 ml (1×107 L-1 ) by ip.Five mice of each group were killed through eye bloodletting at 1,6,12,24,48, 72 h respectively post-infection with Brucella suis S 2 and the peritoneal macrophage were obtained respectively to make smear.Phagocytic rate and phagocytic index were calculated by the Wright Giemsa staining after infected 1 h.TNF-α,IL-12 and IFN-γlevels of serum at different time points were measured by ELISA.The bacterial load of MΦand spleen were measured by coating method.Results:The phagocytic rate and phagocytic index of MΦin APS 3 dose groups were higher than those of the control group ( P<0.05 ).The microbial load of MΦin APS 3 dose groups at 1 h infected by Brucella suis S 2 were significantly higher than those of control,but significantly lower than those of control at 6,12,24,48,72 h after infected by Brucella suis S2.The microbial load of spleen in APS 3 dose groups at 6 h infected by Brucella suis S 2 were significantly higher than those of control ,but significantly lower than those of control at 12,24,48,72h after infected by Brucella suis S2.The concentrations of TNF-α,IL-12 and IFN-γin the serum of APS groups had significantly been improved ( P<0.05 ).Conclusion: APS can promote the activation of MΦin vivo and strengthen the activity of phagocytosis and killing to Brucella suis S 2.APS can promote the secretion of TNF-α,IL-12 and IFN-γof mice,strengthen the cellular immune response of mice to Brucella suis S 2.

Objective:To explore the causes and preventive strategies of adverse reactions of blood transfusion in patients with hematological diseases.Methods:The clinical data of 150 hematological patients who received 5 184 cases of blood transfusion in Beijing No.6 Hospital from January 2014 to December 2017 were retrospectively analyzed. The blood transfusion adverse reaction reporting forms were collected, and the content of the form included the basic information of the patient, diagnosis, blood transfusion type, blood transfusion time, blood transfusion history, pregnancy history, adverse reaction history of blood transfusion as well as detailed records of clinical symptoms. The blood bank summarized the blood transfusion adverse reaction reporting forms every week.Results:A total of 112 cases of adverse reactions occurred in 5 184 cases of blood transfusion, the incidence rate was 2.16%. Sixty-eight cases (1.32%) had anaphylaxis, mainly caused by platelet transfusion, and 44 cases (0.85%) had fever, mainly caused by transfusion of red blood cells suspension, and no other adverse reactions were found. The incidence rates of adverse reactions of blood transfusion in patients with blood transfusion history and anaphylaxis history were significantly higher than those in patients without blood transfusion history and anaphylaxis history, and the differences were statistically significant [2.34% (102/4 350) vs. 1.20% (10/834), χ2 = 6.899, P = 0.009; 3.06% (98/3 200) vs. 1.42% (14/984), χ2 = 7.767, P = 0.005]; the incidence rate of adverse reactions of blood transfusion in female patients was significantly higher than that in male patients, and the difference was statistically significant [2.82% (60/2 126) vs. 1.70% (52/3 058), χ2 = 8.356, P = 0.004]; there was no significant difference in the incidence rate of adverse reactions of blood transfusion between patients < 18 years old and ≥ 18 years old [1.16% (3/259) vs. 2.21% (109/4 925), χ2 = 1.295, P = 0.255]. Conclusions:The main manifestations of adverse reactions of component blood transfusion are anaphylaxis and non-hemolytic fever. Clinical medical staff must carefully control the blood transfusion indications and select the appropriate blood components to reduce the adverse reactions of blood transfusion and ensure the safety of blood transfusion.

Objective To investigate the utility of antibiotics in elderly patients of respiratory department in No. 210 Hospital of PLA, and to understand the rational use of antibiotics in respiratory department. Methods A total of 534 patients >60 in respiratory department of our hospital from September to December in 2011 were randomly selected. The usage rate of antibiotics, the types that used, dosing routes and so on were recorded and analyzed. Some rational suggestions were dis‐cussed put forward. Results ① The usage rate of antibiotics was 97%. ② A total of 21 kinds of antibiotics had been used. The types of drugs are Amp C, levofloxacin, β‐lactamase inhibitors, nitromidazoles, aminoglycoside, antifungals, MALS, lincomycin. The percentage rate of the top three antibiotics were 59. 97%, 17. 30% and 7. 58%. ③ There were some problems in the usage of antibiotics, such as no indication of medicine, high use rate of antibiotics, improper variety selection, dosing routes and drug intervals, irrational use of drugs combinations. Conclusion The application rate of antibiotics was 97%, there was some illegiti‐mate phenomenon in the use of antibiotics. Some attentions should be paid to the rational use of antibiotics in elderly patients, and to raise the rational use rate of antibiotics.

Brucella is an intracellular pathogen that invades a host and settles in its immune cells; however, the mechanism of its intracellular survival is unclear. Modification of small ubiquitin-related modifier (SUMO) occurs in many cellular activities. E2 conjugating enzyme 9 (Ubc9) is the only reported ubiquitin-conjugating enzyme that links the SUMO molecule with a target protein. Brucella's intracellular survival mechanism has not been studied with respect to SUMO-related proteins and Ubc9. Therefore, to investigate the relationship between Brucella melitensis 16M and SUMO, we constructed plasmids and cells lines suitable for overexpression and knockdown of SUMO1 and Ubc9 genes. Brucella 16M activated SUMO1/Ubc9 expression in a time-dependent manner, and Brucella 16M intracellular survival was inhibited by SUMO1/Ubc9 overexpression and promoted by SUMO1/Ubc9 depletion. In macrophages, Brucella 16M-dependent apoptosis and immune factors were induced by SUMO1/Ubc9 overexpression and restricted by SUMO1/Ubc9 depletion. We noted no effect on the expressions of SUMO1 and Ubc9 in B. melitensis 16M lipopolysaccharide-prestimulated mouse RAW264.7 macrophages. Additionally, intracellular survival of the 16M△VirB2 mutant was lower than that of Brucella 16M (p < 0.05). VirB2 can affect expression levels of Ubc9, thereby increasing intracellular survival of Brucella in macrophages at the late stage of infection. Collectively, our results demonstrate that B. melitensis 16M may use the VirB IV secretion system of Brucella to interact with SUMO-related proteins during infection of host cells, which interferes with SUMO function and promotes pathogen survival in host cells.
Animals ; Apoptosis ; Brucella melitensis ; Brucella ; Immunologic Factors ; Macrophages ; Mice ; Plasmids

Objective: To establish the patient derived xenograft (PDX) model of pseudomyxoma peritonei (PMP), and identify the key characteristics of tumor biology of this model, in order to provide a reliable model for studying the pathological mechanisms and new therapeutic strategies of PMP. Methods: PMP tumor tissue was obtained from surgery and cut into pieces after washing. Then tumor pieces were implanted subcutaneously in BAL B/c-nu mice for 6 stable passages. In the 7th passage, tumor tissue was implanted orthotopically into abdomen. Subcutaneous tumor and orthotopic tumor were then homogenized to make tumor cell suspension, implanted into abdomen of 10 BAL B/c-nu mice through midline laparotomy, 100 μl for each. The key experimental parameters including body weight changes in the observation period, experimental peritoneal cancer index (ePCI) score at the autopsy, histopathological and immunohistochemical characteristics, and gene expression profiles by high-throughput whole-genome exon sequencing were detected and recorded. Results: The successful rate of established orthotopic PDX model of human PMP was 100% (10/10). The animals showed smooth body weight increases after tumor inoculation until day 27, then the body weight began to decrease steadily. Widespread tumor dissemination of PMP tumor through the whole abdomen was found by autopsy, including the diaphragm, liver, spleen, stomach, kidney, parietal peritoneum, bowel and mesenterium. Gelatinous ascites was also observed in abdominopelvic cavity. The ePCI score ranged from 5 to 9, with a 8 of median ePCI. Histopathological studies showed peritoneal mucinous carcinomatosis accompanied with signet ring cells (PMCA-S), obvious tumor cell atypia and parenchymal invasion.Immunohistochemistry showed the expressions of MUC1, MUC2, MUC5AC, CEA, CA199, CK20, CDX-2 and Ki-67 were positive, MUC6, CK7 and p53 were negative. Whole-exome sequencing identified that the most significant genetic alteration is the exon10 missense mutation c. 1621A>C of KIT gene, the mutation abundance was 89.7%. Conclusion PDX model of PMCA-S is successfully established, which displays the characters of high-degree malignancy, high proliferation and strong aggressiveness.

Objective: To establish patient derived xenograft (PDX) model of malignant peritoneal mesothelioma (MPM), and to identify the key characteristics of tumor biology of the model, so as to provide an experiment platform for studying the pathologic mechanisms and new therapeutic strategies for MPM. Methods: Surgically excised MPM tumor tissues were inoculated subcutaneously in BALB/c-nu/nu mice for 3 stable passages. In the 4th passage, the subcutaneous tumors were harvested under aseptic conditions, cleaned and made into MPM tumor cell homogenate. Four nude mice (two males and two females) were selected and one male and one female nude mouse were inoculated in the abdominal cavity at the dose of 100 μL, others were inoculated at a dose of 200 μL. The PDX model of MPM was established. The changes of body mass in nude mice were measured regularly, the extent of abdominal and pelvic tumors was judged by experimental peritoneal cancer index (ePCI) score, and the pathologic characteristics of tumors were analyzed. Results: The subcutaneous and abdominal animal models of MPM were successfully established. The subcutaneous tumor model grew into tumor on the 20th day, followed by a slow growth stage between the 20th and 29th day, then a rapid growth stage between the 30th and 57th day. According to the dose of tumor cells (100, 200 μL) and timing (14th and 69th days after grafting), the abdominal tumor model successfully simulated the early and late clinical stages of MPM. The HE staining results of the MPM nude mice model showed that the tumor was epithelial mesothelioma and invaded most of the organs, including liver, spleen, pancreas, mesentery. Immunohistochemical staining for calretinin, cytokeratin 5/6, WT1 and Ki-67 were positive. Whole-genome exon sequencing identified 26 and 36 high frequency gene mutations in tumors derived from the PDX model and clinical sample from patients, including 21 common gene mutations. Conclusions The PDX model of MPM is established. The model is characterized by highly malignant tumor with rapid growth and high invasiveness.

Animals ; Chloroquine/pharmacology* ; Longevity ; Rats