Objective:To construct a VP8-mRNA vaccine using human rotavirus spike protein VP8 domain as the immunogen and analyze its immunogenicity in mice.Methods:The VP8-mRNA sequence was designed, optimized, and synthesized. The VP8 gene of rotavirus G1P[8] type was used to construct the plasmid pUC57-VP8-Kan-SapⅠ, which was then sequenced. The plasmid confirmed by sequencing was subjected to large-scale amplification and extraction, followed by linearization, in vitro transcription, and capping. The purified capped products were encapsulated with lipid nanoparticles using a microfluidic control apparatus. The encapsulated VP8-mRNA vaccine was administered intramuscularly to mice at 10, 15, and 20 μg. Serum samples were collected for antibody detection by ELISA. Cellular immune responses were detected by flow cytometry and ELISPOT. Statistical analysis was performed using one-way or two-way analysis of variance and Tukey-Kramer test. Results:The encapsulated VP8-mRNA vaccine was rounded and spherical, with a particle size of about 100 nm, a polymer dispersion index of 0.088, and an encapsulation rate of 92.3%. Two doses of VP8-mRNA vaccine immunization could induce a good immune response in mice. The level of IgG antibody induced after immunization in the 15 μg group was comparable to that of the 20 μg group, and there was no statistical difference ( P>0.05), but the antibody levels in the two groups were significantly higher than that in the 10 μg group ( P<0.000 1). VP8-mRNA vaccine could induce neutralizing antibodies against rotavirus G1 and G9 types. The highest level of neutralizing antibodies against rotavirus type G1 was observed in the 15 μg group, which was significantly higher than that in the 10 μg group ( P<0.05). All immunization groups exhibited good neutralizing ability against rotavirus G9 type. The results of ELISPOT showed that lymphocytes from mice in each vaccine group were able to secrete IFN-γ when stimulated with VP8 peptide. Flow cytometry showed that the proportions of CD8 + T cell subsets in the vaccine groups were higher than that in the control group. Conclusion:The VP8-mRNA vaccine has good immunogenicity in mice and can induce good humoral and T-cell immune responses.

Objective:To construct a VP8-mRNA vaccine using human rotavirus spike protein VP8 domain as the immunogen and analyze its immunogenicity in mice.Methods:The VP8-mRNA sequence was designed, optimized, and synthesized. The VP8 gene of rotavirus G1P[8] type was used to construct the plasmid pUC57-VP8-Kan-SapⅠ, which was then sequenced. The plasmid confirmed by sequencing was subjected to large-scale amplification and extraction, followed by linearization, in vitro transcription, and capping. The purified capped products were encapsulated with lipid nanoparticles using a microfluidic control apparatus. The encapsulated VP8-mRNA vaccine was administered intramuscularly to mice at 10, 15, and 20 μg. Serum samples were collected for antibody detection by ELISA. Cellular immune responses were detected by flow cytometry and ELISPOT. Statistical analysis was performed using one-way or two-way analysis of variance and Tukey-Kramer test. Results:The encapsulated VP8-mRNA vaccine was rounded and spherical, with a particle size of about 100 nm, a polymer dispersion index of 0.088, and an encapsulation rate of 92.3%. Two doses of VP8-mRNA vaccine immunization could induce a good immune response in mice. The level of IgG antibody induced after immunization in the 15 μg group was comparable to that of the 20 μg group, and there was no statistical difference ( P>0.05), but the antibody levels in the two groups were significantly higher than that in the 10 μg group ( P<0.000 1). VP8-mRNA vaccine could induce neutralizing antibodies against rotavirus G1 and G9 types. The highest level of neutralizing antibodies against rotavirus type G1 was observed in the 15 μg group, which was significantly higher than that in the 10 μg group ( P<0.05). All immunization groups exhibited good neutralizing ability against rotavirus G9 type. The results of ELISPOT showed that lymphocytes from mice in each vaccine group were able to secrete IFN-γ when stimulated with VP8 peptide. Flow cytometry showed that the proportions of CD8 + T cell subsets in the vaccine groups were higher than that in the control group. Conclusion:The VP8-mRNA vaccine has good immunogenicity in mice and can induce good humoral and T-cell immune responses.

Analyzing the distribution characteristics of allergen sIgE in the serum of patients with respiratory and skin mucosal diseases in Shanghai City, and to provide epidemiological characteristics and diagnostic basis for the prevention and treatment of allergic respiratory and dermo-mucous diseases in Shanghai City. Adopting cross-sectional research, a total of 3 822 patients who received treatment in Tongren Hospital, Shanghai Jiao Tong University School of Medicine from July 2022 to July 2023 due to respiratory diseases or skin and dermo-mucous symptoms were included. Among them, there were 1 456 males and 2 366 females, with an age range of 1-97 years old. The median age (interquartile range) was 33 (27, 44) years old. The sIgE was detected by using immunoblotting. Statistical analysis was conducted using SPSS 22.0 software, and the comparison of count data (rates) between groups was conducted using χ 2 test. The results showed that a total of 3 377 (88.4%) cases among 3 822 patients were at least one allergen sIgE positive, and 72.9% (2 788/3 822) of them were multiple allergies sIgE positive. The top five allergen sIgE positive rates were Dermatophagoides pteronyssinus (37.9%, 1 447/3 822), Dermatophagoides farinae (32.1%, 1 225/3 822), milk (31.7%, 1 211/3 822), fungi (28.3%, 1 080/3 822), and Blomia tropicdis (23.8%, 909/3 822), with only milk was a kind of food allergen. The highest positive rates within the respiratory system disease group or dermo-mucous disease group were also these five allergens, without any difference in disease categories. The positive rates of cat dandruff, Humulusscandens, and juniper/birch in the respiratory system disease group were significantly higher than those in the skin and mucous membrane disease group, while the positive rates of shrimp/crab were relatively low (11.3% vs 14.9%, χ 2=9.616, P=0.002). Whether in the respiratory system disease group or the dermo-mucous disease group, the positive rates of Dermatophagoides pteronyssinus in male patients were significantly higher than those of females(42.6% vs 35.7%,41.0% vs 34.4%), with statistical significance ( χ 2=12.515, P<0.001; χ 2=5.143, P=0.023), And the three allergens, Dermatophagoides farinae, cat dander, and egg white allergens are also characterized by this feature.In addition, the positive rates of milk(33.8% vs 30.1%, χ 2=3.911, P=0.048), shrimp/crab(13.2% vs 10.0%, χ 2=6.423, P=0.014) in the respiratory system disease group were higher in males than in females, while in the dermo-mucous disease group, dog dander(20.5% vs 14.6%, χ 2=6.726, P=0.010) and peanuts/soybeans(10.5% vs 6.9%, χ 2=4.698, P=0.030) showed this phenomenon. In both the respiratory system disease group and the dermo-mucous disease group, there were 6 types of inhaled allergens (Dermatophagoides pteronyssinus, Dermatophagoides farinae, Blomia tropicdis, cat dandruff, dog dander, fungi) and 4 types of food allergens (egg yolks, egg white allergens, milk, shrimp/crab). However, the positivity rate of Aspergillus fumigatus (7.2% vs 9.3% vs 10.5% vs 15.7%, χ 2=10.996, P=0.012)in the respiratory disease group and cockroaches(4.2% vs 11.3% vs 9.6% vs 16.4%, χ 2=10.237, P=0.017) in the skin and mucosal disease group was the lowest in the underage group. There are seasonal differences in the positivity rates of allergens, with most allergens having significantly higher positivity rates in summer and autumn. In conclusion, the most common allergens sIgE positive in patients with respiratory and dermo-mucous diseases in Shanghai City are Dermatophagoides pteronyssinus, Dermatophagoides farinae, milk, fungi, and Blomia tropicdis. The trend of allergen sIgE prevalence in the two major categories of diseases is basically consistent. Allergen sIgE distribution varies among patient populations of different gender, age or season, and clinical prevention and treatment can be based on the results of serum allergen testing.

【Objective】 To explore the feasibility of blood transfusion compatibility testing for multiple myeloma(MM) patients treated with anti-CD38 monoclonal antibody daratumumab (DARA) after DARA-Fab fragment blocking, and to evaluate the transfusion efficacy by comparing with dithiothreitol(DTT) method. 【Methods】 After DARA was prepared into DARA-Fab fragments using PierceFab preparation kit, the neutralization effects of different volumes (5, 10, 15, 30 μL) on screening cells and panel cells were confirmed. DARA-Fab fragments and screening cells with specific antigens and corresponding monoclonal antibody reagents were used as the experimental group and the control group with the same volume of saline for incubating and centrifugin.Twenty MM patients treated with DARA were selected for cross-matching with DARA-Fab and DTT respectively, and the laboratory indexes before and after transfusion were statistically analyzed, and the two blood matching methods were compared. 【Results】 After incubating and centrifuging, the results of DARA-Fab fragments(15, 30 μL) with screening cells and serum mixed with DARA were negative, while those of DARA-Fab(5, 10 μL) were positive. 15μL DARA-Fab treated antibody identification cells (2, 3, 4, 5, 7, 9, 11) were negative, antibody identification cells (1, 6, 8, 10, 12) were negative after 30 μL DARA-Fab fragments treatment; the results of MNS, Duffy, Kidd, Kell, Lewis, Rh blood group system of the experimental group were consistent with those of the control group; the hemoglobin (Hb) (g/L) of 20 patients after infusion of RBC (73.90±1.90) was significantly higher than that before transfusion (63.60±1.58), P<0.01. There was no significant difference in total bilirubin(TBil)(μmol/L)(16.25±3.54 vs 17.87±3.57), direct bilirubin(DBIL)(μmol/L)(6.31±2.32 vs 7.10±2.80)and indirect bilirubin(I-Bil)(9.94±1.38 vs 10.77±1.22) before and after infusion(P>0.05).And no statistical difference was noticed in Hb (10.75±1.04 vs 10.30±0.98), TBil (3.31±1.47 vs 3.31±0.55), DBIL(2.76±1.24 vs 2.60±0.83), and I-Bil(1.97±0.40 vs 2.82±0.53) between the DTT treatment method and the DARA Fab fragment treatment before and after transfusion(P>0.05). 【Conclusion】 DARA-Fab can remove the interference of RBC on cross matching by blocking CD38 antigen. This method has no effect on the antigens of common RBC blood group systems, and shows significant blood transfusion efficacy as that of DTT method.

Analyzing the distribution characteristics of allergen sIgE in the serum of patients with respiratory and skin mucosal diseases in Shanghai City, and to provide epidemiological characteristics and diagnostic basis for the prevention and treatment of allergic respiratory and dermo-mucous diseases in Shanghai City. Adopting cross-sectional research, a total of 3 822 patients who received treatment in Tongren Hospital, Shanghai Jiao Tong University School of Medicine from July 2022 to July 2023 due to respiratory diseases or skin and dermo-mucous symptoms were included. Among them, there were 1 456 males and 2 366 females, with an age range of 1-97 years old. The median age (interquartile range) was 33 (27, 44) years old. The sIgE was detected by using immunoblotting. Statistical analysis was conducted using SPSS 22.0 software, and the comparison of count data (rates) between groups was conducted using χ 2 test. The results showed that a total of 3 377 (88.4%) cases among 3 822 patients were at least one allergen sIgE positive, and 72.9% (2 788/3 822) of them were multiple allergies sIgE positive. The top five allergen sIgE positive rates were Dermatophagoides pteronyssinus (37.9%, 1 447/3 822), Dermatophagoides farinae (32.1%, 1 225/3 822), milk (31.7%, 1 211/3 822), fungi (28.3%, 1 080/3 822), and Blomia tropicdis (23.8%, 909/3 822), with only milk was a kind of food allergen. The highest positive rates within the respiratory system disease group or dermo-mucous disease group were also these five allergens, without any difference in disease categories. The positive rates of cat dandruff, Humulusscandens, and juniper/birch in the respiratory system disease group were significantly higher than those in the skin and mucous membrane disease group, while the positive rates of shrimp/crab were relatively low (11.3% vs 14.9%, χ 2=9.616, P=0.002). Whether in the respiratory system disease group or the dermo-mucous disease group, the positive rates of Dermatophagoides pteronyssinus in male patients were significantly higher than those of females(42.6% vs 35.7%,41.0% vs 34.4%), with statistical significance ( χ 2=12.515, P<0.001; χ 2=5.143, P=0.023), And the three allergens, Dermatophagoides farinae, cat dander, and egg white allergens are also characterized by this feature.In addition, the positive rates of milk(33.8% vs 30.1%, χ 2=3.911, P=0.048), shrimp/crab(13.2% vs 10.0%, χ 2=6.423, P=0.014) in the respiratory system disease group were higher in males than in females, while in the dermo-mucous disease group, dog dander(20.5% vs 14.6%, χ 2=6.726, P=0.010) and peanuts/soybeans(10.5% vs 6.9%, χ 2=4.698, P=0.030) showed this phenomenon. In both the respiratory system disease group and the dermo-mucous disease group, there were 6 types of inhaled allergens (Dermatophagoides pteronyssinus, Dermatophagoides farinae, Blomia tropicdis, cat dandruff, dog dander, fungi) and 4 types of food allergens (egg yolks, egg white allergens, milk, shrimp/crab). However, the positivity rate of Aspergillus fumigatus (7.2% vs 9.3% vs 10.5% vs 15.7%, χ 2=10.996, P=0.012)in the respiratory disease group and cockroaches(4.2% vs 11.3% vs 9.6% vs 16.4%, χ 2=10.237, P=0.017) in the skin and mucosal disease group was the lowest in the underage group. There are seasonal differences in the positivity rates of allergens, with most allergens having significantly higher positivity rates in summer and autumn. In conclusion, the most common allergens sIgE positive in patients with respiratory and dermo-mucous diseases in Shanghai City are Dermatophagoides pteronyssinus, Dermatophagoides farinae, milk, fungi, and Blomia tropicdis. The trend of allergen sIgE prevalence in the two major categories of diseases is basically consistent. Allergen sIgE distribution varies among patient populations of different gender, age or season, and clinical prevention and treatment can be based on the results of serum allergen testing.

Objective To study the application of 99Tcm in rabbit cerebral thromboembolic stroke and thrombolysis effect of recombinant tissue plasminogen activator (rt-PA).Methods The 0.5 mL radioactive pertechnetate sodium (specification:5 mCi/2mL and radiation intensity 92.5 MBq/mL) was combined with 30 μL stannous chloride (5 mg/mL),and the 20 μL mixture was joined to whole blood,red blood cells,and plasma for labelling.Then 50 μL CaCl2 (0.5 mol/L) and bovine thrombin (50 IU/mL) were doped in mixture,and rapidly sucked into a polyethylene plastic pipe (PE80).Thrombus was formed for 2 h at 37 ℃ and cut into small pieces of 10 mm.Autologous blood clots combined with 99Tcm from external carotid artery were injected to internal carotid artery of rabbit,the radioactivity (counts per minute,CPM) was measured by gamma counting instrument,and the improvement of rt-PA 4.5 mg/kg (clinical equivalent dose) on this model was observed.Results After thromboembolism,CPM increased approximately by (5.1 ± 1.3) times,which suggested that the model was reliable.The rt-PA 4.5 mg/kg had significant progressive thrombolysis effect.Conclusion 99Tcm tracer technology could be applied to rabbit cerebral stroke model,which is stable and reliable

At present,the network media reports of Chinese medical disputes appeared blowout growth,and also brought the loss of propriety,the loss of balance,the loss of justice and other ethical issues while it played the role of public opinion supervision in the medical disputes.In view of these,from the concept definition of ethics anomie in network media report of medical disputes,this paper analyzed and interpreted the ethics anomie in network media report of medical disputes,pointed out existing problems and further put forward creating the network media professional ethics norms,establishing a social supervisory system which integrates self-discipline and heteronomy,building a doctor-patient docking platform and other perfecting countermeasures.

Objective:To investigate the effect of IL-17A on the differentiation and maturation of murine bone marrow-derived dendritic cells( BMDCs ) . Methods: Murine bone marrow cells were isolated and cultured in RPMI1640 complete medium in the presence of GM-CSF(20 ng/ml) for 8 days to induce differentiation of murine bone marrow cells to DC progenitors. Then these cells were treated with LPS(1 μg/ml) for 36 h which polarized immature DCs into mature DCs. Different concentrations of rmIL-17A(10 or 100 ng/ml) was added to the culture medium at different stages of BMDC differentiation and maturation. Co-stimulatory molecules expression on BMDC were analyzed by flow cytometry,and the culture supernatants were analyzed for IL-12p40 and IL-10 level by ELISA. Results:rmIL-17 could promote co-stimulatory molecules( CD40,CD80,CD86 and MHCⅡ) expression on BMDCs in a does-dependent manner,especially,the expression of CD40 and MHCⅡhad a significant increase in high concentration of rmIL-17A group;rmIL-17A was added while LPS induced maturation of BMDCs. CD40,CD80,CD86 and MHCⅡexpression on BMDC increased sharply in LPS plus rmIL-17A stimulation group,besides,CD86,MHCⅡ showed a higher level expression on BMDC with the increase of con-centration of rmIL-17A. Furthermore,secretion of IL-12p40 and IL-10 increased significantly in the group of DCs treated with LPS plus low concentration of rmIL-17 compared with the group without rmIL-17(P<0. 001). However,high concentration of rmIL-17A group showed significantly higher levels of IL-12p40(P<0. 001),but there was no difference in IL-10. Conclusion:IL-17A promotes the phe-notypic development of BMDC progenitors propagated in GM-CSF and cooperate with LPS to induce BMDC differentiation and matura-tion.

Objective To investigate the existence of neglect and explore the relationship between spacial attentional deficits and hemisphere in patients with schizophrenia. Methods 51 schizophrenic patients and 55 healthy controls were measured with line bisection test and number bisection test、 Results In the Line Bisection Test,shcizophrenic patients group showed a negative deviation, in which the 2cm line( -0.03 ± 0. 74 )% and mean ( - 0. 02 ± 0. 03 ) % deviation rates were more significantly leftward than that of the normals ( ( 0. 00 ±0.02 ) %, ( - 0.00 ± - 0.01 ) %, P ＜ 0. 05 ) respectively. The mean deviation rates of the patients (P ＜ 0. 01 ) and control groups (P ＜ 0. 05 ) were also significantly leftward compared with zero value. In the number bisection test,schizophrenic patients group showed a negative deviation and a significant leftward bias on the interval 5 ( - 0.07 ±0.18)% ,7( -0.08 ±0.22)% and 9( -0.09 ±0.28)% ) than that of the normals(( -0.01 ±0.05)% ,(0.01± 0. 08 ) % and (0.00 ± 0.14 ) %, P ＜ 0.05 respectively). Conclusion Schizophrenic patients have a right-neglect, suggesting that spacial attentional deficits exist, and it may be related to the left cerebral hemisphere dysfunction.

Objective To investigate influence of exogenous insulin in all-in-one parenteral nutrition on blood glucose in infants with very low birth weight (VLBW). Methods Forty-two infants with VLBWI admitted to the department of pediatrics of Xuzhou Hospital affiliated to Southeast University during September 2005 to March 2009 were randomly assigned to Group Ⅰ ( n = 13 ) with exogenous insulin added to all-in-one parenteral nutrition at infusion rate of 0.4 U·kg-1·h-1,GroupⅡ(n = 13) with exogenous insulin at infusion rate of 0.1U·kg-1·h-1 and Group Ⅲ (n = 16) with no exogenous insulin added.Their blood glucose was monitored every two hours. Chi-square test was used for comparing difference in blood glucose abnormality between the three groups and association between blood glucose levels at admission and during hospitalization was analyzed with Spearman correlation. Results Incidence of hyperglycemia and hypoglycemia was 10. 9 percent (29/265) and 18. 1 percent (48/265) in Group Ⅰ, 20. 8 percent (59/284) and 14. 1 percent (40/284) in Group Ⅱ , and 20. 5 percent (61/298) and 11.7 percent (35/298) in Group Ⅲ, respectively. There was significant difference in incidence of hyperglycemia between Groups Ⅰ and Ⅱ ( x2 = 9. 844, P = 0. 002 ) and between Groups Ⅰ and Ⅲ ( x2 = 9. 478, P = 0. 002 ), but no significant difference in it between Groups Ⅱ and Ⅲ ( x2 = 0. 008, P = 0. 928 ). There was significant difference in incidence of hypoglycemia between Groups Ⅰ and Ⅲ ( x2 = 4. 526, P =0. 033 ), but no significant difference in it between Groups Ⅰ and Ⅱ (x2 =1.653, P=0. 199) or between Groups Ⅱ and Ⅲ (x2 =0.709, P =0.400).No significant correlation between endogenous blood insulin level at admission and during hospitalization( r = 0. 082, P = 0. 661 ) was found. Conclusions Blood glucose in infants with VLBW can not be regulated timely by their endogenous insulin itseff. Exogenous insulin added to all-in-one parenteral nutrition at infusion rate of 0. 1 U · kg-1 · h-1 may not significantly reduce incidence of hyperglycemia,while incidence of hypoglycemia can be reduced by exogenous insulin at infusion rate of 0. 4 U · kg- 1 · h -1 that can increase incidence of hypoglycemia Therefore, exogenous insulin is not recommended to be prophylactically added to all-in-one parenteral nutrition for infants with VLBW.