1.Preparation and immunogenicity evaluation of rotavirus VP8-mRNA vaccine
Qingmei LENG ; Xianqiong TANG ; Rong CHEN ; Xiaoqing HU ; Xiaopeng SONG ; Yan LI ; Jinmei LI ; Lida YAO ; Xiaochen LIN ; Jinyuan WU ; Maosheng SUN ; Hongjun LI ; Yan ZHOU
Chinese Journal of Microbiology and Immunology 2025;45(9):727-732
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.
2.Preparation and immunogenicity evaluation of rotavirus VP8-mRNA vaccine
Qingmei LENG ; Xianqiong TANG ; Rong CHEN ; Xiaoqing HU ; Xiaopeng SONG ; Yan LI ; Jinmei LI ; Lida YAO ; Xiaochen LIN ; Jinyuan WU ; Maosheng SUN ; Hongjun LI ; Yan ZHOU
Chinese Journal of Microbiology and Immunology 2025;45(9):727-732
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.
3.Perceived stress and mobile phone addiction among nursing college students: the chain-mediating role of anxiety and flow experience
Shuiqing RONG ; Zhaonan YANG ; Lida YANG ; Qiongyi WANG ; Yanjie YANG ; Zhengxue QIAO ; Xiaohui QIU ; Siyuan KE ; Jiawei ZHOU ; Xiaomei DU ; Wei DUAN ; Yizhi WANG ; Xiuxian YANG
Chinese Journal of Behavioral Medicine and Brain Science 2024;33(6):539-543
Objective:To explore the chain mediating effect of anxiety and flow experience on perceived stress and mobile phone addiction in nursing college students.Methods:In December 2021, a cross-sectional design survey was conducted on 4 179 freshmen and sophomores in a nursing college in Heilongjiang Province. The Chinese perceived stress scale, generalized anxiety disorder-7, flow state scale, and mobile phone addiction tendency scale were selected separately to assess perceived stress, anxiety symptoms, flow experience and mobile phone addiction. SPSS 26.0 software was used for descriptive analysis, independent sample t-test, Spearman correlation analysis, and AMOS 24.0 software was used for mediating effect test. Results:(1) Among the 3 050 nursing students, there were 714(23.41%) students who were addicted to mobile phones. (2) Spearman correlation analysis indicated that perceived stress(27.31±9.56) was positively correlated with anxiety(7.00(1.00, 10.00), r=0.441, P<0.05), flow experience((12.00±3.40), r=0.517, P<0.05), and mobile phone addiction((42.42±13.05), r=0.476, P<0.05).Anxiety was positively correlated with flow experience ( r=0.430, P<0.01) and mobile phone addiction ( r=0.538, P<0.01).Flow experience was positively correlated with mobile phone addiction ( r=0.490, P<0.01). (3) Anxiety and flow experience played seperate mediating and chain mediating roles between perceived stress and mobile phone addiction, accounting for 26.06%(0.165/0.633), 23.54%(0.149/0.633) and 3.48%(0.022/0.633) of the total effect. Conclusion:Perceived stress not only directly affects the mobile phone addiction of nursing students, but also indirectly affects mobile phone addiction through the independent and chain mediating effects of anxiety and flow experience.
4.Analysis of the distribution characteristics of allergen sIgE detection in patients with respiratory and skin mucosal diseases in a hospital in Shanghai City from 2022 to 2023
Binbin XUAN ; Meiyu TAN ; Hanxiao SUN ; Jiajie CHEN ; Lida ZHOU ; Huanhuan ZHANG ; Jiameng YAO ; Yajie WANG ; Jinpiao LIN ; Huiming SHENG
Chinese Journal of Preventive Medicine 2024;58(12):1902-1911
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.
5.Analysis of the distribution characteristics of allergen sIgE detection in patients with respiratory and skin mucosal diseases in a hospital in Shanghai City from 2022 to 2023
Binbin XUAN ; Meiyu TAN ; Hanxiao SUN ; Jiajie CHEN ; Lida ZHOU ; Huanhuan ZHANG ; Jiameng YAO ; Yajie WANG ; Jinpiao LIN ; Huiming SHENG
Chinese Journal of Preventive Medicine 2024;58(12):1902-1911
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.
6.Engineering progress in microbial production of polyhydroxyalkanoates.
Kai YUAN ; Weiqiang ZHOU ; Chao PENG ; Tang TANG ; Qi WANG ; Weitao TANG ; Tai AN ; Bo CHEN ; Haijun LIU ; Lida WU ; Yi LI ; Yi TONG
Chinese Journal of Biotechnology 2021;37(2):384-394
Polyhydroxyalkanoates (PHAs) are polymers obtained by esterification of hydroxy fatty acid monomers. Due to similar mechanical characteristics of traditional petroleum-based plastics, 100% biodegradability and biocompatibility, PHAs are considered to be one of the most potential green materials. However, the application and promotion of PHAs as a green and environmentally friendly material are difficult because of the high production costs. This article focuses on the current methods to reduce production cost of PHAs effectively, such as cell morphology regulation, metabolic pathway construction, economic carbon source utilization and open fermentation technology development. Despite most research results are still limited in laboratory, the research methods and directions provide theoretical guidance for the industrial production of economic PHAs.
Fermentation
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Industry
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Petroleum
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Plastics
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Polyhydroxyalkanoates
7.Study on the application of high-frequency ultrasound to diagnosis of early rheumatoid arthritis
Liang WU ; Hong MA ; Lida XIAO ; Liyi ZHOU
Chongqing Medicine 2015;(18):2493-2495
Objective To study the application value of high frequency ultrasound to the diagnosis of early rheumatoid arthri‐tis .Methods 40 cases were selected ,who were diagnosed with early rheumatoid arthritis ,as the RA group ,another 40 healthy per‐sons were selected as the normal control group .Objects in the 2 groups were examined with high frequency ultrasound and X‐ray . Comparison of the examination results was made ,at the same time ,the results of suprapatellar bursa effusion ,synovial hyperplasia and bone erosion of the positive detection rate in group RA from the analysis of high‐frequency ultrasound and X‐ray examination were also compared .In addition ,the relativity between the related index and ESR index and CRP index in group RA was also ana‐lysed .Results The suprapatellar bursa effusion ,synovial hyperplasia ,bone erosion of color flow rate and bone erosion of the posi‐tive detection rate were higher than those in normal control group(P<0 .05);Compared with the control group ,the artery resist‐ance index was less ,the femoral condyle and lateral condylar cartilage thickness of RA group increased more(P<0 .05) .High fre‐quency ultrasound positive detection rate suprapatellar bursa effusion ,synovial hyperplasia ,bone erosion in patients of group RA were higher than those of X‐ray(P<0 .05) .The knee joint suprapatellar bursa effusion ,synovial hyperplasia of synovial membrane thickenss ,the thickness of the color blood flow grade and CRP ,ESR of patients in RA group were correlated(P<0 .01) .Conclusion High frequency ultrasound can respond to the situation of early rheumatoid arthritis patients ,and the lesion detection rates are higher ,there is a certain correlation between ultrasonographic indexes and CRP ,ESR .
8.Antifibrotic effect of TY501 on bleomycin-induced pulmonary fibrosis in rats and related mechanism
Yanyan GENG ; Bing YU ; Zhixing ZHOU ; Qianqian HU ; Weiren XU ; Lida TANG
Chinese Pharmacological Bulletin 2015;(2):210-215
Aim To investigate TY501′s role in bleo-mycin ( BLM )-induced pulmonary fibrosis in rats. Methods Forty rats were randomly divided into 5 groups, including the sham operation, BLM, PFD, TY501(high and low dose) groups. After administra-tion of BLM intratracheally, PFD and TY501 were giv-en in each group daily, according to the dosage de-signed during 21 days. Lung coefficient, PaO2 were tested before killing the rats. The contents of ALB, ALP, LDH, GSH, HYP were detected by regent kit respectively. PCⅢ and COL4 were determined by ELISA. Results ( 1 ) Some indicators of alveolitis in early stage of IPF: the contents of lung coefficient in three treatment groups were lower and PaO2 was higher than those in BLM group ( P<0. 05 ); compared with BLM group, the contents of ALB, ALP, LDH in the treatment groups reduced on 21 st day ( P<0. 05 );the expression of GSH in BLM group was increased for feedback regulation and higher than the treatment groups and the sham operation group (P<0. 05);(2) some indicators of pulmonary interstitial fibrosis in late stage of IPF:the expressions of HYP, PCⅢand COL4 were reduced after the treatment. There were signifi-cant differences compared with BLM group ( P <0. 05 ) . Conclusions TY501 is valuable for the ther-apy of IPF, the same as the positive drug pirfenidone. TY501 attenuates BLM-induced pulmonary fibrosis, which may be related to the affection of TGF-βpathway and inhibition of MMPs.
9.Medical service,medical insurance and the regulatory model in Germany and the inspirations
Yizhong ZHOU ; Xiaohong CAO ; Lida LI ; Yingyao CHEN
Chinese Journal of Hospital Administration 2012;28(5):396-400
The paper briefly introduced the outpatient and inpatient medical services and medical insurance system in Germany.It also covered the regulatory model and its structure and role of the German medical service,which are characterized ofSociety-led self-governance.Based on the observation and inspiration of this model,the authors came up with.recommendations to improve China' s medical service,medical insurance and regulatory model.
10.Influence of insulin in all-in-one parenteral nutrition on blood glucose level in infants of very low birth weight
Xiangyu GAO ; Yang CHEN ; Yingjun SUN ; Lida LI ; Shenying QU ; Zhaojun WEI ; Guangling ZHOU ; Ying ZHOU ; Bo YANG ; Peizhi HAO
Chinese Journal of General Practitioners 2010;09(10):695-699
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.

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