Objective: To explore the value of nomogram based on DCE-MRI radiomics combined with clinical-ra- diological features in predicting HR status in breast cancer with Her-2 low expression． Methods: A total of 198 pa- tients of Her-2 low expression breast cancer who underwent standardized breast MRI in our hospital from January 2019 to February 2025 were retrospectively analyzed．Patients were divided into HR ( + ) group ( n = 137) and HR ( －) group ( n = 61) ．The cases were divided into a training set ( 138 cases) and a testing set ( 60 cases) in a 7 ∶ 3 ratio．Clinical-radiological model was based on clinical and traditional radiological features，radiomics model was based on DCE-MRI，and combined model was constructed，respectively．The nomogram was drawn，and re- ceiver operating characteristic curve was used to compare the performance of different models in predicting HR sta- tus． Results: The DCE-MRI radiomics score ( Radscore) between the HR ( + ) group and the HR ( －) group showed statistical differences in both the training and testing sets ( both P＜0. 001) ．The AUC of the clinical-radio- logical model based on lesion mobility，Ki67，TIC type，enhancement pattern and maximum diameter for predicting HR status in the training set and testing set were 0. 643 and 0. 616，respectively．The AUC of the DEC-MRI ra- diomics model in the training set and testing set were 0. 897 and 0. 860，respectively．The nomogram drawn by combining clinical-radiological features and Radscore showed better predictive performance in both the training set ( AUC = 0. 913) and testing set ( AUC = 0. 898) than the clinical-radiological model ( all P＜0. 05) ． Conclusion The nomogram combined by DCE-MRI radiomics and clinical-radiological features can effectively predict HR sta-tus of breast cancer with low Her-2 expression，which is helpful to the building of individualized treatment plan for breast cancer patients．

Objective To study the immunoadjuvant effects of chitosan oligosaccharide(COS),including the immune activation and the triggering of lysosomal escape,and to explore whether COS can be used as an adjuvant for attenuated live bacteria vector vaccines.Methods 1)Mouse macrophages RAW264.7 cells were cultured with COS at 0 mg/mL(the control group)and 0.1-4 mg/mL for 24 h and the effect on cell viability was measured by CCK8 assay.Mouse macrophages RAW264.7 were treated with COS at 0(the control group),1,2,and 4 mg/mL for 24 h.Then,the mRNA expression levels of the cytokines,including IFN-γ,IL-10,TGF-β,and TLR4,were determined by RT-qPCR assay.2)RAW264.7 cells were treated with 1 mL of PBS containing different components,including calcein at 50 μg/mL,COS at 2 mg/mL,and bafilomycin A1,an inhibitor,at 1 μmol/mL,for culturing.The cells were divided into the Calcein group,Calcein+COS group,and Calcein+COS+Bafilomycin A1 group accordingly.Laser scanning confocal microscopy was used to observe the phagocytosis and the intracellular fluorescence distribution of calcein,a fluorescent dye,in RAW264.7 cells in the presence or absence of COS intervention to determine whether COS was able to trigger lysosomal escape.3)LM?E6E7 and LI?E6E7,the attenuated Listeria vector candidate therapeutic vaccines for cervical cancer,were encapsulated with COS at the mass concentrations of 0.5 mg/mL,1 mg/mL,2 mg/mL,4 mg/mL,and 8 mg/mL.Then,the changes in zeta potential were measured to select the concentration of COS that successfully encapsulated the bacteria.Phagocytosis of the vaccine strains by RAW264.7 cells was measured before and after LM?E6E7 and LI?E6E7 were coated with COS at 2 mg/mL.Results 1)CCK8 assays showed that,compared with the findings for the control group,the intervention of RAW264.7 cells with COS at different concentrations for 24 h was not toxic to the cells and promoted cell proliferation,with the difference being statistically significant(P＜0.05).According to the RT-qPCR results,compared with those of the control group,the COS intervention up-regulated the mRNA levels of TLR4 and IFN-γ in RAW264.7 cells,while it inhibited the mRNA expression levels of TGF-β and IL-10,with the most prominent effect being observed in the 4 mg/mL COS group(P＜0.05).2)Laser scanning confocal microscopy revealed that the amount of fluorescent dye released from lysosomes into the cells was greater in the Calcein+COS group than that in the Calcein group.In other words,a greater amount of fluorescent dye was released from lysosomes into the cells under COS intervention.Furthermore,this process could be blocked by bafilomycin A1.3)The zeta potential results showed that COS could successfully encapsulate the surface of bacteria when its mass concentration reached 2 mg/mL.Before and after the vaccine strain was encapsulated by COS,the phagocytosis of LM?E6E7 by RAW264.7 cells was 5.70%and 22.00%,respectively,showing statistically significant differences(P＜0.05);the phagocytosis of LI?E6E7 by RAW264.7 cells was 1.55%and 6.12%,respectively,showing statistically significant differences(P＜0.05).Conclusion COS has the effect of activating the immune response of macrophages and triggering lysosomal escape.The candidates strains of coated live attenuated bacterial vector vaccines can promote the phagocytosis of bacteria by macrophages.Further research is warranted to develop COS into an adjuvant for bacterial vector vaccine.

Objective:To investigate the changes of inflammation and immune function in children with chronic tonsillitis after tonsillotomy. Methods:Prospectively collected 60 children with obstructive sleep apnea （OSA） diagnosed as chronic tonsillitis with adenoids and tonsillar hypertrophy from January to June 2021. Two groups were divided, the experimental group （n=30） underwent bilateral partial tonsillectomy + adenoidectomy by hypothermia plasma ablation, and the control group （n=30） underwent adenoidectomy by using the same hypothermia plasma ablation method. The number of tonsillitis attacks before surgery and within one year after surgery was recorded, and the serum immunoglobulin IgM, IgG, IgA, complement C3 and complement C4 levels before operation, one month and three months after operation were measured. Results:The number of tonsillitis attacks in the experimental group and the control group at one year after surgery was lower than that before surgery（P<0.05）; The number of inflammatory attacks in the experimental group was （0.50±0.63） times/year, which was lower than that of （1.33±0.80） times/year in the control group. There was no significant difference in the five immunization results of the two groups at one month and three months after operation compared with before operation, and there was also no significant difference between the experimental and the control groups. Conclusion:Partial tonsillectomy can be applied to children with chronic tonsillitis, which can effectively reduce the number of tonsillitis attacks and has no effect on the immune function of children.
Child ; Humans ; Tonsillectomy/methods* ; Hypothermia ; Tonsillitis/surgery* ; Adenoidectomy ; Palatine Tonsil/surgery* ; Inflammation ; Chronic Disease ; Immunity

Objective To construct Listeria monocytogenes(LM)and Listeria ivanovii(LI)balanced lethal systems expressing cervical cancer antigens,to study their basic biological characteristics,and to provide reference data for the immunotherapy of cervical cancer.Methods Through seamless cloning via in vitro ligation kit,the HPV16 E6E7 fusion protein antigen gene constructed in our lab was spliced to the complement plasmid pCWgfp-LM dal-Amp that contained the nutritional gene dal.Then,we replaced the ampicillin(Amp)resistance gene of the complement plasmid with the asd nutrition gene.The ligation reaction mixture was transformed into Escherichia coli(E.coli)recipient bacteria DH5аΔasd and the complement plasmid pCWgfp-E6E7-LM dal-Ampfree,which expressed cervical cancer antigens and had no Amp resistance,was obtained by nutrition screening from the E.coli DH5аΔasd.The plasmid pCWgfp-E6E7-LM dal-Ampfree was complemented into LMΔdd and LIΔdd,the attenuated nutrition-deficient Listeria strains with the virulence genes actA and plcB and nutrition genes dal and dat deleted by electroporation,thereby obtaining LM and LI balanced lethal systems expressing cervical cancer antigen genes.The in vitro growth of the strains was observed.Western blot was performed to examine the status of antigen protein expression.PCR was performed to measure the in vitro passage stability of complement plasmid pCWgfp-E6E7-LM dal-Ampfree.Their basic biological characteristics were examined by biochemical reaction tests and hemolysis assay.Results Two Listeria balanced lethal systems expressing cervical cancer antigen were successfully constructed.The HPV16 type E6E7 fusion protein was successfully expressed in the two Listeria balanced lethal systems.pCWgfp-E6E7-LM dal-Ampfree,the positive plasmid expressing cervical cancer antigen,maintained stable existence in the two Listeria balanced lethal systems.The two Listeria balanced lethal systems expressing cervical cancer antigen showed significantly better recovery growth in comparison with Listeria nutrition deficiency strains.The results of biochemical reaction tests showed that most of the biochemical reaction of the two Listeria balanced lethal systems expressing cervical cancer antigen were consistent with those of Listeria attenuated strains.The two Listeria balanced lethal systems expressing cervical cancer antigen still maintained the hemolytic ability,although their hemolytic ability was slightly inferior to that of the Listeria balanced lethal systems not expressing cervical cancer antigen and the Listeria attenuated strains.Conclusion The two Listeria balanced lethal systems expressing cervical cancer antigen genes are constructed successfully.They display normal in vitro growth.The complement plasmid pCWgfp-E6E7-LM dal-Ampfree can maintain stable existence in vitro,showing little change in its biochemical characteristics and hemolytic ability.Further research should be conducted to investigate the potential of these two recombinant strains to be used as candidate strains for cervical cancer therapeutic vaccine.

Objective: To analyze the changes in the expression of hypoxia inducible factor-1α (HIF-1α) and inflammatory cytokines and to investigate the role of HIF-1α in regulating the production of inflammatory cytokines during influenza A (H1N1) virus infection. Methods: BALB/c mice were injected with H1N1 virus to establish the mouse model of H1N1 virus infection. Fifteen BALB/c mice were randomly divided into three groups: control group, H1N1 virus group and H1N1 virus+ HIF-1α inhibitor group. Inflammatory cytokines (IL-6, TNF-α, IL-1β and IL-10) in samples of serum and lung tissues were detected by Luminex and ELISA. Levels of HIF-1α in serum and lung tissue samples were detected by Western blot and ELISA, respectively. Results: Compared with the control group, the levels of inflammatory cytokines in serum (IL-6, TNF-α, IL-1β and IL-10) and lung tissues (IL-6 and TNF-α) and the expression of HIF-1α in serum and lung tissues in the H1N1 virus group were significantly increased. The levels of HIF-1α, IL-6, TNF-α IL-1β and IL-10 in lung tissues in H1N1 virus+ HIF-1α inhibitor group were significantly lower than those of the H1N1 virus group. Conclusion During H1N1 virus infection, the levels of inflammatory cytokines and HIF-1α were significantly increased. The production of inflammatory cytokines was significantly reduced after inhibiting HIF-1α expression, suggesting that HIF-1α might promote the production of inflammatory cytokines.

OBJECTIVE To explore the role of adenoidectomy in the treatment of pediatric secretory otitis media.METHODS 120 cases of children diagnosised with secretory otitis media and adenoid hypertrophywho came to our department between January 2013 and May 2015 were devided into 4 groups,30 cases in each group.Group 1 only got drug therapy;Group 2 got adenoidectomy combined with drug therapy;Group 3 got paracentesis or tympanostomy tube;Group 4 got paracentesis or tympanostomy tube,with adenoidectomy at the same time.RESULTS 1.The difference incurative time and recurrence rate were statistically significant between group 1 and group 2.2.There was no statistical difference in healing time but statistical difference in recurrence rate between group 3 and group 4.CONCLUSION For children with secretory otitis media and adenoid hypertrophy,adenoidectomy can shorten the cure time and reduce the recurrence rate at the same time.

Objective: To establish fluorescence quantitative polymerase chain reaction (PCR) to detect integrated HIV DNA in peripheral blood mononuclear cells. Methods: A total of 30 HIV-seropositve individuals were enrolled in this study, including 10 subjects with a detection limit of 20 copies/ml of plasma, 10 patients with drug resistance and 10 patients with no history of antiretroviral therapy (ART). Cultivated ACH2 cells carried a single copy of the integrated HIV genome. We have built pMD19T-CD3 plasmid and calculated the copy number. We used oligonucleotides ULF1 specific for the long terminal repeats (LTR) regions and two oliligonucleotides specific for human Alu sequences to pre-amplified the integrated HIV DNA. Samples and serial dilutions of ACH2 cells were all pre-amplified, the products of which were used for the second round fluorescence amplifications. The Lambda T primers, UR2 primers and HIV Taqman probes were used for second round amplifications in integrated HIV DNA assay. The CD3IN5 primers, CD3IN3 primers and CD3 Taqman probes were used for CD3 quantification. Results: Serial 5-fold dilutions of the plasmid were used as standards for CD3 gene quantifications. The equation of the linear regression was y=-2.731x+ 43.01(R²=0.953). The cellular input was quantified by number of human genome equivalents (CD3 gene, 2 copies/cell). The copies of ACH2 cells was 4.271×10^4, which was the cellular input of the ACH2 cells. Serial 5-fold dilutions of ACH2 cells were used to generate a standard curve for the integrated HIV DNA assays. The equation of the linear regression was y=-3.146x+ 39.11 (R²=0.968). The ratio between the number of copies of the integrated HIV DNA form and the number of cells (CD3 copies) was calculated to obtain the frequency of cells. Based on the test between different groups, each group had no difference (P> 0.05). Conclusions This method presented advantage in detection of the lower copies of virus. It hinted that the current antiretroviral therapy can not effectively attack against latent viral reservoirs. It also provided a basis for new therapeutic intervention.

OBJECTIVETo analyze virulence genes and molecular characteristics of Vibrio parahaemolyticus isolated from sporadic cases with diarrhea in tow sentinel hospitals of Shanghai, 2010-2012.

METHODSA total of 2 729 stool samples were collected from two surveillance sentinel hospitals in Shanghai 2010-2012. Vibrio parahaemolyticus strains were isolated and identified from diarrhea out patients using TCBS agar plates and biochemical reactions. Thermostable direct hemolysingene (tdh), thermostable-related hemolysin gene (trh), hemolysin gene (tlh) were detected by multiplex PCR method. Isolates were analyzed by PFGE and MLST. The PFGE profiles were analyzed using BioNumerics software.

RESULTSA total of 30 clinical Vibrio parahaemolyticus strains isolated from 2 729 stool samples. The anually Vibrio parahaemolyticus isolation rate during 2010 to 2012 were 1.1%(11/973), 1.0%(11/1 120) and 1.3%(8/636) respectively. The PCR positive rates of virulence genes tlh, tdh and trh were 100%, 97% and 0 respectively. The Vibrio parahaemolyticus strains were divided into 13 PFGE types (P1-P13)and 3 ST types (ST-189, ST-799, ST-3). Among 13 PFGE types, P4 was the main PFGE type, accounting for 30%(9/30). P9, P10 were accounting for 12% (4/30) respectively, P1, P2, P12, P13 were accounting for 7%(2/30) respectively, the others types were 3%(1/30) respectively. MLST analysis results showed there are three ST types, ST3 was 84%(25/30), ST189 and ST799 were accounting for 13% (4/30) and 3% (1/30) respectively.

CONCLUSIONThe infection rate of Vibrio parahaemolyticus was not very high from 2010-2012 in Shanghai, all strains were positive for tlh and negative for trh. ST3 was the major type of Vibrio parahaemolyticus.

China ; Diarrhea ; Genotype ; Hemolysin Proteins ; Hospitals ; Humans ; Multilocus Sequence Typing ; Outpatients ; Polymerase Chain Reaction ; Sentinel Surveillance ; Vibrio Infections ; Vibrio parahaemolyticus ; Virulence

Objective To ensure the safety of oral administration at night among elderly inpatients. Methods By the purposive sampling method, a total of 165 clinical nurses and 274 elderly patients were selected as sample. Elderly patients received nocturnal tracking and 165 nurses underwent survey questionnaire, that investigate risk factors for night safety of oral-administration. Results For circumstances of night oral medicine administration among 165 nurses, the rate of medicine inspection was highest (90. 9%), while the implementation rate of putting medicine into mouth was lowest (33. 3%). Drug regimen and administration time of elderly patients had positive correlation with the implantation level of putting medicine in the mouth by nurses (r=0.407, 0. 335;P < 0. 05). The compliance and posture of medication administration had positive correlation with medicine knowledge education to patients by nurses (r=0. 380, 0. 429;P<0. 05), and the adverse reactions of patients was positively correlated with nurses observed after taking the drug (r=0. 464,P<0. 05). Conclusions We should focus on guiding elderly patients to take medicine correctly and compliance of medicine order, and carry out the training of comprehensive drug knowledge for nurses to decrease the risks of night medicine administration.

OBJECTIVETo comprehensively analyze the clinical characteristics, treatment strategies and outcome of patients with thrombotic thrombocytopenic purpura (TTP).

METHODSA retrospective survey of 51 TTP patients confirmed in our database. Relevant statistical analyzes were performed by GraphPad Prism 5 software.

RESULTS51 cases of patients with acquired TTP were identified as idiopathic TTP. In our study, only 18 cases (35.29%) had typical pentalogy of TTP, where thrombocytopenia (100.00%), microangiopathic hemolytic anemia (92.16%) and neurologic abnormalities (88.24%) were more common than fever (72.55%) and renal abnormalities (70.59%). Plasma ADAMTS13 activity was detected in 37 patients with TTP with ADAMTS13 deficiency confirmed in 31 patients (83.78%). Plasma exchange with response of 72.3% was still the preferred strategy in TTP with individuation. Among 36 survival TTP patients, 8 patients (22.22%) relapsed. 15 patients (29.41%) died in our study. The mean ages of responders and deaths were of (37.5±14.5) and (50.1±18.9) respectively; whereas total bilirubin level of responders and deaths were of (43.3±23.5)μmol/L and (63.7±39.7) μmol/L respectively, the differences were statistically significant. Conversely, body temperature, WBC, HGB, PLT, serum creatinine and LDH showed no significant differences (P>0.05).

CONCLUSIONThe diagnosis of TTP was based on comprehensive analysis of clinical manifestations. Plasma ADAMTS13 activity test had a higher clinical practical value. The therapeutic alliance with corticosteroids, immunosuppressive agents and Rituximab significantly improved its outcome. The age and high total bilirubin level at onset were associated with less sensitive to plasmapheresis and poor prognosis.

ADAM Proteins ; blood ; ADAMTS13 Protein ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Male ; Middle Aged ; Plasma Exchange ; Purpura, Thrombotic Thrombocytopenic ; diagnosis ; therapy ; Retrospective Studies ; Young Adult