In order to explore the role of BspE protein in Brucella infection,yeast two-hybrid tech-nique was used to screen host cell proteins that interact with BspE protein.The constructed BspE recombinant plasmid pGBKT7-BspE was used as bait plasmid to hybridize with the RAW264.7-cD-NA library of mouse mononuclear macrophages by yeast two-hybridization technique.The positive clones were extracted by plasmid,sequenced and co-immunoprecipitation to determine the host cell proteins that could interact with BspE.The subcellular localization of BspE proteins was analyzed by confocal laser microscopy.The physical and chemical properties,protein structure and function of BspE interacting proteins were analyzed by bioinformatics.The siRNA for one of the BspE inter-acting proteins was synthesized,the expression of its gene was silenced in HEK293T cells,and the silenced cells was infected with Brucella M5-90 and the number of intracellular bacteria was coun-ted.The results showed that the decoy plasmid pGBKT7-BspE was successfully constructed,and the plasmid could express BspE protein in yeast.Eight positive clones were obtained from the host cell genome library by yeast two-hybridization.The positive clones were identified as RBM27 and PCBP1 by sequencing,backcross and co-immunoprecipitation.Bioinformatics was used to predict the cell location,protein structure and amino acid composition of RBM27 and PCBP1.After siRNA interference,the expression level of PCBP1 was significantly decreased and the amount of M5-90 in the cell was increased.Brucellosis secreted protein BspE interacts with host proteins RBM27 and PCBPl,and PCBP1 negatively regulates the proliferation of Brucellosis.

Objective: To investigate the current situation of risk assessment of neonatal pressure injury (PI) and using of the assessment scale from 8 hospitals in Zhejiang Province,providing reference for further improvement of the scale. Methods: A cross-sectional study was used to investigate 184 nurses in the department of neonatal intensive care unit admitted to 8 hospitals in Zhejiang Province by using a self-made questionnaire. Results: Three of the eight hospitals did not use scale for neonatal PI assessment routinely. Of the remaining five hospitals, three hospitals used Neonatal Skin Risk Assessment Scale (NSRAS), one hospital used Braden Q Pediatric Skin Risk Assessment Scale (Pediatric Braden Q Scale) and one hospital used Neonatal/Infant Braden Q scale (Neonatal/Infant Braden Q Scale). 129 nurses from 5 hospitals evaluated the scales routinely used by the department.Nearly 93.80% (121/129) of the nurses thought the scales were easy to understand, 76.74% (99/129) of the nurses thought the scales were easy to measure, 76.74% (99/129) of the nurses thought the scales were good at predicting, 75.19% (97/129) of the nurses thought the scales were suitable for newborns, 37.21% (48/129) of the nurses thought the scales did not adequately assess the severity of the condition, 34.11% (44/129) of the nurses thought the scales did not adequately assess the medical device factors, 24.81% (32/129) of the nurses thought the scales scores were not clearly defined. Conclusion The use of risk assessment scale for neonatal PI has not been unified yet, and the three scales need to be improved. The department should pay attention to the risk assessment of newborn PI and strengthen the awareness of PI prevention.

Objective To investigate the prokaryotic expression and immunoreactivity of BspE,a type Ⅳ secretion protein of Brucella,and the effect of recombinant protein BspE on cytokines.Methods According to the BspE gene of Brucella M5-90 published in GenBank,the gene fragments were synthesized by a company and then ligated into PUC57 vector for sequencing.The sequenced gene was cloned into a prokaryotic expression vector pET-28α and transformed.Induced expression was performed in E.coli DE3 competent cells.The obtained target protein was purified by a Ni-NTA affinity column,and its reactogenicity was analyzed by Western blotting.Mouse RAW264.7 cells were treated with 25 g/L BspE recombinant protein for 12,24,48 h,and the control group was treated with the same amount of BSA instead of BspE,and enzyme-linked immunosorbent assay (ELISA) was used to detect interleukin (IL)-1β level.Results The recombinant expresed plasmid of pET-28α-BspE was successfully obtained.The results of Western blotting showed a single band with a relative molecular mass of about 30.1 × 103,and the recombinant protein BspE had good reactogenicity,and IL-1β levels (ng/L)were significantly elevated by the recombinant protein BspE (12 h:43.27 ± 2.13 vs 30.24 ± 1.66,24 h:57.78 ± 3.44 vs 41.22 ± 1.22,48 h:72.52 ± 3.04 vs 46.77 ± 2.75,t =8.38,7.86,10.89,P ＜ 0.05).Conclusions BspE recombinant protein has better immunoreactivity and can increase the expression level of IL-1β in mouse macrophages.This study provides a scientific basis for the role of effector proteins in the pathogenesis of Brucella.

Objective By studying clinical features, treatment and prognosis of eosinophilic gastroenteritis of infants resulted from milk protein allergy an, to improve the diagnosis and treatment level of eosinophilic gastroenteritis. Methods 24 cases of infants which diagnosed eosinophilic gastroenteritis were chosen from June of 2010 to January of 2014 in children’s Hospital of XX province and By retrospective analysis clinical manifestations, endoscopic features, histopathology, treatment and prognosis of the 24 cases. Results The 24 cases who were vomiting, paroxysmal crying, abdominal distension (100.00%), which accompanied by haematemesis 23 cases (95.83%), 1case (4.17%) hematochezia, 17cases (70.83%) eczema, 21 cases (87.50%) mild to moderate anemia, 1 cases (4.17%) severe anemia, 19 cases (79.17%) the increasing of peripheral blood eosinophil cells, 8 cases (33.33%)the increasing of IgE of the serum and 4 cases (16.67%) the test of antibody of the Helicobacter pylori in Serum was positive; 3 cases (12.50%) were milk protein allergy by the detecting of food allergen-speciifc IgE antibodies, the endoscopic characteristics were hyperemia, edema, erosion, ulcer of gastric and duodenal mucosa. Among them, 24 cases (100.00%) were gastritis, 5 cases (20.83%) duodenitis and 1 cases (4.17%) duodenal ulcer. The histopathology of the 24 cases revealed that there were gastric or duodenal eosinophils infiltration (> 20/HPF) and were all associated with mast cell infiltration; By antisecretory, protection of the gastrointestinal mucosa and the obviating of milk protein had a satisfactory treatment effect, 24 cases of children with oral general formula milk test conifrmed that the milk protein allergy, The 3 cases of the patients were reviewed by 8~12 weeks after gastroscope, and the mucosa of the duodenum was smooth, Eosinophils were/HPF<8, mast cells were/HPF<5. Conclusion There are no speciifc clinical and endoscopic manifestations in eosinophilic gastroenteritis of infants resulted from milk protein allergy, gastrointestinal mucosa eosinophil inifltration and simultaneously are accompanied by abnormal mast cell inifltration;Mucosal type without the use of corticosteroids, through milk protein avoidance treatment can achieve satisfactory results, But definite diagnosis must rely on biopsy and eosinophils, combined with avoidance stimulation test of milk protein can further confirmed, But the excitation test should be at least 10 days of observation of children, and carefully recorded symptoms, so as not to delay the missed diagnosis of CMPA.

Objective To construct a scoring system of combination of procalcitonin and C-reactive protein for diagnosis of invasive bacterial diarrhea in children. Methods The clinical data of hospitalized children with acute diarrhea were retrospectively analyzed. All of the children were divided into two groups, invasive bacterial diarrhea group and nonbacterial diarrhea group. The scoring system of combination of two markers for diagnosis of invasive bacterial diarrhea in children was constructed by means of two categories logistic regression analysis using procalcitonin and C- reactive protein as parameters. In addition,receiver operating characteristic curve(ROC)was used to evaluate the diagnostic accuracy.Results One hundred ten cases of invasive bacterial diarrhea and 108 cases of nonbacterial diarrhea were included. The levels of serum procalcitonin and C-reactive protein in invasive bacterial diarrhea group were significantly higher than those in nonbacterial diarrhea group, and there were statistical difference(P all<0.05).The area under the ROC curve(AUC)of invasive bacterial diarrhea was 0.894 when the established scoring system was used, which was higher than the AUCs when either serum procalcitonin or in C- reactive protein was used (P<0.05). The best diagnostic boundary value for combination of serum procalcitonin and C- reactive protein scoring was 0.52 with a sensitivity at 80.9% and specificity at 88.9%. Conclusions The scoring system of combination of procalcitonin and C- reactive protein has good accuracy in diagnosis of invasive bacterial diarrhea in children, and can assist the early diagnosis of the disease.

In this paper, we analyze the status of the medical device testing and the situation of the unqualifie varieties by sorting out the situation of the national medical device supervision and inspection from 2013 to 2016. We analyze the results of the inspection deeply, and find that the safety risk points exists in the quality of the medical device testing products and management system, product technology requirements, the national standards, industry standards or other aspects, then explain one by one with typical examples. At the same time the corresponding suggestions are put forward to production companies and regulators respectively on the existing problems.

Objective To evaluate the clinical effect of popliteal artery perforator-based sural neurovascular flap for repairing soft tissue defects in middle and distal upper leg.Methods Between December,2011 to September,2015,18 cases with skin soft tissue defects on the middle and distal upper leg were treated with popliteal artery perforator-based sural neurovascular flap,in which 12 cases were males,and 6 cases were females.The age was from 24 to 55 years,with the average age of 35.9 years.The size of tissue defects ranged from 3.0 cm × 3.0 cm to 7.0 cm × 9.0 cm.Results All flaps survived completely in 18 cases,1 case of marginal infection heated after dressing,and the rest cases in Ⅰ healing.The outline and function of survived flap were satisfactory during 3-16 months follow-up,with two-point discrimination of 4.0-10.0 mm.Conclusion The anterograde sural neurovascular flap based on the popliteal artery perforator provides a practical option for covering tissue defects in middle and distal upper leg.This flap is characteristiced by reliable blood supply without sacrificing main vesses,good contour and texture,in addition,the operation is easy of handling.

Objective:To investigate the effects of TGF-β1 on T lymphocytes of BALB/c mice infected with Echinococcus granulosus( E.granulosus ) in vitro.Methods: The inhibitor group:the spleen cells of BALB/c mouse were co-cultured with E.granulosus and SB525334.The control group:the spleen cells of BALB/c mouse were co-cultured with E.granulosus and PBS.The blank group:the spleen cells of BALB/c mouse were co-cultured with RPMI-1640 medium and SB525334.The lymphocytes were collected at 48 h post-infection.The T lymphocyte subsets, the number of CD4+CD25+T cells, the number of NK cells, and the expression of NKG2D receptor were detected by flow cytometry.The NK cell activity was determined with the lactate dehydrogenase leakage assay(LDH).Results:The inhibition the TGF-β1 receptors resulted in the increase of in the number of CD4+T cells,the decrease in the number of CD8+T cells,the increase of in the ratio CD4+/CD8+T cells,the decrease of in the number of CD4+CD25+T cells,the increase in the expression of the NKG2D receptors,the increase in the lysis rate of Yac-1 cells by NK cells,and a positive cor-relation between the expression of activity receptor NKG2D and killing activity of NK, which were mediated by E.granulosus.Conclusion: The inhibition of TGF-β1 receptors can enhance the immune response of T lymphocytes against E.granulosus infection in vitro.

Helicobacter pylori (H.pylori) infection is the most important pathogenic microorganisms associated with the development of chronic gastritis, peptic ulcer and gastric mucosa-associated lymphoid tissue lymphoma (MALT) in children. H.pylori infection may occur in early childhood and last a lifetime even directly affecting the health of adulthood unlessH.pylori infection was eradicated. This article aims to review the infection condition, infection pathway and immunological characteristics ofH.pylori and its relationship with extragastroduodenal diseases in children.

PURPOSE: In the present study, we investigated the incidence of cardiotoxicity within 5 years of trastuzumab treatment and evaluated potential risk factors in clinical practice. METHODS: The study cohort included 415 patients diagnosed with early breast cancer (EBC). Cardiotoxicity incidence was evaluated in patients receiving trastuzumab and those who did not. Multivariate Cox proportional hazards regression models were used to estimate hazard ratios and 95% confidence intervals of potential risk factors for trastuzumab-related cardiotoxicity after appropriate adjustments. RESULTS: Incidence of cardiotoxicity in patients treated with trastuzumab was significantly higher than that in controls (23.7% vs. 10.8%, p<0.001). This result was adjusted for factors that might increase the risk of cardiotoxicity, such as history of coronary artery diseases or the use of anthracyclines for more than four cycles. CONCLUSION: Our findings indicated that treatment with trastuzumab was strongly associated with cardiotoxicity in EBC patients.
Anthracyclines ; Breast Neoplasms* ; Cohort Studies ; Coronary Artery Disease ; Humans ; Incidence ; Observational Study* ; Prospective Studies* ; Risk Factors ; Trastuzumab