1.Identification of sulfakinin and its receptor gene in Aedes aegypti and transcriptomic changes in genes knockdown strains
JIANG Linlong ; ZHU XiaoJing ; ZHANG Lei ; HUANG Yuqi ; XU Jingyun ; HAN Qian
China Tropical Medicine 2024;24(1):75-
Objective By exploring the function of sulfakinin (SK) and sulfakinin receptor (SKR) of Aedes aegypti, it laid a certain experimental basis and theoretical basis for the research and development of new insecticides targeting neuropeptides and their receptors. Methods This study investigated the roles of SK and its receptor gene in Ae. aegypti using bioinformatics analysis and Clustered Regularly Interspaced Short Palindromic Repeats(CRISPR)/Cas9 knockout technology. Subsequently, RNA interference technology was employed to suppress the expression of SK or its receptor in adult mosquitoes. Lastly, transcriptome sequencing technology was utilized to identify and analyze differentially expressed genes between the interference group and the control group in order to gain insights into their functions. Results It was found that there is only one SK receptor in Ae. aegypti. In addition, during the construction of mutant strains of Ae. aegypti SK and its receptor gene, it was found that only 2% of the G0 generation mutant strains mutated to form chimeras, with a large number of male chimeras dying, and only 14% of female chimeras being able to lay eggs, ultimately resulting in no effective G1 generation mutants. Transcriptome data showed, compared to the control group, 181 genes were significantly differentially expressed after interfering with the SK gene, with 62 genes significantly upregulated and 119 genes significantly downregulated. In addition, after interference with the sulfakinin receptor, 110 genes exhibited significant differential expression, including 20 upregulated and 90 downregulated genes. Cross-analysis of the two datasets identified 46 genes with significant expression changes after interference with sulfakinin or its receptor, with only 4 genes upregulated and the remaining 42 genes significantly downregulated, and the differentially expressed genes were mainly enriched in the metabolic pathway, endocrine system, and digestive system. Conclusions The SK and its receptor gene are highly conserved and may primarily play roles in regulating the energy metabolism and digestion functions in Ae. aegypti, thus playing an important role in regulating insect growth and development.
2.Isolation and cultivation of HIV strains circulating in China and the screening and identification of the standard strains
Siqi LI ; Guoxin ZHU ; Lin LI ; Jingyun LI ; Hanping LI ; Yongjian LIU ; Lei JIA ; Xiaolin WANG ; Tianyi LI ; Bohan ZHANG ; Jingwan HAN ; Chuntao MA
Chinese Journal of Experimental and Clinical Virology 2023;37(3):286-295
Objective:To screen the HIV standard strains with typical biological characteristics of HIV strains circulating in China through the isolation, culture, genotype and phenotype identification of HIV from the whole blood samples of HIV-infected persons, confirm genetic characteristics, traceability, and in line with the Standard Strains of Pathogenic Microorganism-technical Specifications for Establishment of HIV Strains (T/CPMA 027—2023).Methods:Whole blood samples were collected from 48 HIV infected patients. Peripheral blood mononuclear cells (PBMCs) were isolated from the samples and co-cultured with PBMCs isolated from healthy persons′ whole blood samples to isolate and culture HIV from infected persons. We determined concentration of p24 antigen and the virus titer in the culture supernatant. The viral RNA was extracted from the successfully isolated strains, and the gag, pol genes and env C2V3 fragments of the viral genome were amplified and sequenced. The genotype, gene recombination and drug resistance sites were determined according to the viral gene sequences. Virus infection and replication were monitored by inoculating the virus culture supernatant into Ghost cells expressing CCR5 or CXCR4 to determine the viral tropism.The formation of syncytium was observed by inoculating the virus culture supernatant into MT-2 cells to determine whether was a syncytium-induced phenotype. Results:Fourteen strains with p24 antigen concentration > 1 ng/ml in culture supernatant were isolated and cultured from 48 fresh EDTA anticoagulated whole blood samples of HIV infected persons. Of the 14 strains, only one strain with a titer≥10 5 TCID 50/ml, 8 strains with titers ≥10 4 TCID 50/ml, and the other 5 strains with titers≥10 3 TCID 50/ml. Phylogenetic analysis showed that the genotypes of the strains were 9 strains of subtype B, 3 strains of CRF01_AE and 2 strains of CRF07_BC recombinant. Genotypic resistance analysis showed that 11 strains contained drug resistance sites. Ghost cells were used to verify the tropism of the strains, and it was found that 8 strains were CCR5 tropism, 6 strains were CXCR4 & CCR5 dual tropism. Only 2 of the 14 strains could induce MT-2 cytopathic effect, which was syncytium-inducing phenotype. Conclusions:Fourteen HIV strains with typical biological and genetic characteristics were isolated to screen the standard HIV strains. Among which, 1 strain was evaluated as a standard HIV strain that meets the Standard Strains of Pathogenic Microorganism-technical Specifications for Establishment of HIV Strains (T/CPMA 027—2023). This study can also provide technical guidance for the screening of the HIV standard strains. Next step is to complete the application and reserve database construction according to the sharing mechanism of the HIV standard strains, to provide resources for the researches of HIV vaccines and drugs.
3.Phenotypic resistance properties of HIV-1 CRF01_AE strain main drug resistance mutations to integrase inhibitors
Tengchong YAO ; Jingwan HAN ; Hanping LI ; Lei JIA ; Xiaolin WANG ; Lin LI ; Jingyun LI
Chinese Journal of Microbiology and Immunology 2022;42(2):81-87
Objective:To analyze the effects of the main drug resistance mutations in the integrase (IN) region on the resistance of HIV-1 CRF01_AE strains, and compare the differences with subtype B strains.Methods:Seven IN region mutations or combined mutations (T66K, F121Y, Q148K, N155H, G118R, R263K, Q148K/N155H) were selected from the HIV drug resistance database of Stanford University in the United States, and introduced to the IN region of HIV-1 B subtype infectious clone pNL4-3 and CRF01_AE infectious clone pGX002 by seamless cloning, homologous recombination and point mutation. The mutant plasmids were transfected into 293T cells for virus packaging. The culture was expanded in MT2 cells and infectious titers were detected. Half maximal inhibitory concentrations (IC 50) of four integrase inhibitors (INSTIs), raltegravir (RAL), elvitegravir (EVG), dolutegravir (DTG) and bictegravir (BIC), against 14 mutant viruses were detected and compared with the IC 50 against the wild-type viruses. Results:B subtype and CRF01_AE plasmids carrying seven IN region mutations or combined mutations were successfully constructed, and 14 recombinant viruses were packaged with an infectious titer of 10 4-10 6 median tissue culture infective dose (TCID 50)/ml. The recombinant viruses replicated efficiently in MT2 cells. The concentrations of HIV-1 p24 antigen contained in the supernatants of cell culture reached 830-2 700 ng/ml. Five mutations or combined mutations (T66K, F121Y, Q148K, N155H, Q148K/N155H) caused CRF01_AE and B subtype strains to be highly resistant to RAL and EVG, resulting in an increase in the IC 50 by 200 times and 2 000 times or more as compared with the IC 50 against the wild-type viruses. The same mutation-caused fold changes of IC 50 of RAL and EVG against CRF01_AE were significantly lower than that of subtype B ( P<0.01). Q148K/N155H mutation caused B subtype and CRF01_AE to be highly resistant to DTG and BIC, with IC 50 increased by more than 50 times. Other mutations had little effects on the sensitivity to DTG and BIC. Conclusions:Fourteen HIV-1 strains carrying seven INSTI resistance mutations based on B subtype and CRF01_AE were constructed. Five mutations resulted in high resistance to RAL and EVG, and there was a high level of cross-resistance. Resistance to RAL and EVG caused by the same mutation was higher in B subtype than in CRF01_AE. The combined mutation of Q148K and N155H was associated with greater resistance to DTG and BIC, indicating that the genetic barrier of DTG and BIC resistance was high. DTG and BIC could effectively inhibit the strains carrying INSTI resistance mutations without obvious subtype difference.
4.Qualitative study on the psychological status and nursing needs of patients with maintenance hemodialysis
Zhengyan LI ; Rui DING ; Jingyun BAI ; Rui HAN ; Yuanyuan HU ; Xue HAN ; Jinhong MIAO
Chinese Journal of Modern Nursing 2022;28(24):3256-3259
Objective:To understand the psychological status and nursing needs of maintenance hemodialysis (MHD) patients and provide reference for clinical targeted nursing.Methods:Using the phenomenological research method, a total of 15 patients with MHD in the First Affiliated Hospital of Zhengzhou University were selected as the research objects by the purposive sampling method, and they were given semi-structured individualized interviews. The Colaizzi 7-step analysis method was used to organize the data, extract the inductive themes and analyze the results.Results:The psychological status of MHD patients was repressed fear, despair and helplessness and acceptance. The nursing needs of MHD patients included disease-related knowledge needs, social support needs and health guidance needs.Conclusions:MHD patients have negative psychology. Medical staff need to understand the psychological status and nursing needs of patients and formulate targeted nursing interventions to meet the actual needs of patients and improve the quality of life of them.
5.Effect of dietary management based on family participation in elderly maintenance hemodialysis patients
Zhengyan LI ; Jinhong MIAO ; Jingyun BAI ; Yufeng LEI ; Xue HAN ; Yuanyuan HU
Chinese Journal of Modern Nursing 2022;28(30):4225-4230
Objective:To explore the effect of family-participated dietary management on dietary compliance behavior and nutritional status in elderly maintenance hemodialysis (MHD) patients.Methods:From July 2019 to December 2020, a total of 120 elderly MHD patients admitted in the First Affiliated Hospital of Zhengzhou University were selected as the research objects, and divided into the experimental group and the control group with 60 cases in each group by random number table method. The control group conducted routine dietary management, and the experimental group received dietary management based on family participation. Six months after intervention, the dietary compliance attitude, dietary compliance behavior and nutritional status were compared between the two groups.Results:Six months after intervention, the scores of social restriction attitude, self-care attitude, acceptance attitude and dietary compliance behavior attitude of the experimental group were higher than those of the control group, and the differences were statistically significant ( P<0.05) . The scores of fluid restriction behavior, potassium intake behavior, self-care compliance, compliance in the face of difficulties, and dietary compliance of the experimental group were higher than those of the control group, and the differences were statistically significant ( P<0.05) . The serum prealbumin (PA) , albumin (ALB) , hemoglobin (Hb) , and transferrin (TRF) of the experimental group were higher than those of the control group, and the differences were statistically significant ( P<0.05) . Conclusions:Diet management based on family participation can enhance the attitude of dietary compliance, promote the development of dietary compliance behavior, and improve the nutritional status of elderly patients with MHD.
6.Effects of salvianolic acid B on the migration and tube formation of high glucose-induced retinal vascular endothelial cells and the underlying mechanism
Kunxiu JIANG ; Huihui SUN ; Xingzhuo SONG ; Xirui WANG ; Qianqian SUN ; Jingyun TIAN ; Hongli LI ; Yonggang LIU ; Jing HAN
International Journal of Traditional Chinese Medicine 2020;42(11):1094-1101
Objective:To observe the effects of salvianolic acid B on migration and tube formation of the retinal vascular endothelial cell (RVEC) in high glucose, and explore its mechanism with network pharmacology.Methods:The cells were divided into normal group, model group and 1.0, 0.5, 0.1 μg/ml salvianolic acid B group according to the random number table method. The cells of each group were added with 5.5 mmol/L glucose for intervention, and the salvianolic acid B group was added with 1.0, 0.5, and 0.1 μg/ml salvianolic acid B for intervention. After 72 h, the cell viability of each group was detected by the CCK-8 method. The cells were divided into normal group, model group and low-, medium-, and high-dose salvianolic acid B group according to the random number table method. Then the cells of the normal group were added with 5.5 mmol/L glucose; the model group was added with 25 mmol/L glucose; the low-, medium-, and high-dose salvianolic acid B group was added with 25 mmol/L glucose and 0.062 5, 0.1250, 0.250 0 μg/ml salvianolic acid B. Then by taking Transwell test to detect the number of cell migration, and Matrigel test to analyze the total length of cells tubes. The active targets of Salvianolic acid B were screened by SuperTarget and Swiss TargetPrediction. Then, the targets of diabetic retinopathy were obtained by searching the GAD database, pharmGkb database, TTD database, DiGSeE database and OMIM database. The effective targets of drug-disease interaction were screened, and the component-target-disease interaction network was constructed by Cytoscape. Finally, the effective targets were analyzed by DAVID for GO analysis and KEGG pathway enrichment analysis. Molecular docking was performed by using Accelrys Discovery Studio Client 2.5 software.Results:The CCK-8 method showed that the cell absorbance values of 0.5 and 0.1 μg/ml salvianolic acid B group were not significantly different from those of the normal group ( P>0.05). The results of Transwell experiment and Matrigel experiment showed that compared with the model group, the relative number of migrating cells and the total length of tubule formation in each dose group of salvianolic acid B decreased ( P<0.05 or P<0.01). The interaction network revealed that salvianolic acid B acted on 46 targets and 8 signaling pathways. Conclusions:Salvianolic acid B could inhibit the migrating and tube forming ability of RVEC cultivated by high glucose. The results suggest that salvianolic acid B may play roles in preventing diabetic retinopathy.
7.Chinese Society of Allergy Guidelines for Diagnosis and Treatment of Allergic Rhinitis.
Lei CHENG ; Jianjun CHEN ; Qingling FU ; Shaoheng HE ; Huabin LI ; Zheng LIU ; Guolin TAN ; Zezhang TAO ; Dehui WANG ; Weiping WEN ; Rui XU ; Yu XU ; Qintai YANG ; Chonghua ZHANG ; Gehua ZHANG ; Ruxin ZHANG ; Yuan ZHANG ; Bing ZHOU ; Dongdong ZHU ; Luquan CHEN ; Xinyan CUI ; Yuqin DENG ; Zhiqiang GUO ; Zhenxiao HUANG ; Zizhen HUANG ; Houyong LI ; Jingyun LI ; Wenting LI ; Yanqing LI ; Lin XI ; Hongfei LOU ; Meiping LU ; Yuhui OUYANG ; Wendan SHI ; Xiaoyao TAO ; Huiqin TIAN ; Chengshuo WANG ; Min WANG ; Nan WANG ; Xiangdong WANG ; Hui XIE ; Shaoqing YU ; Renwu ZHAO ; Ming ZHENG ; Han ZHOU ; Luping ZHU ; Luo ZHANG
Allergy, Asthma & Immunology Research 2018;10(4):300-353
Allergic rhinitis (AR) is a global health problem that causes major illnesses and disabilities worldwide. Epidemiologic studies have demonstrated that the prevalence of AR has increased progressively over the last few decades in more developed countries and currently affects up to 40% of the population worldwide. Likewise, a rising trend of AR has also been observed over the last 2–3 decades in developing countries including China, with the prevalence of AR varying widely in these countries. A survey of self-reported AR over a 6-year period in the general Chinese adult population reported that the standardized prevalence of adult AR increased from 11.1% in 2005 to 17.6% in 2011. An increasing number of original articles and imporclinical trials on the epidemiology, pathophysiologic mechanisms, diagnosis, management and comorbidities of AR in Chinese subjects have been published in international peer-reviewed journals over the past 2 decades, and substantially added to our understanding of this disease as a global problem. Although guidelines for the diagnosis and treatment of AR in Chinese subjects have also been published, they have not been translated into English and therefore not generally accessible for reference to non-Chinese speaking international medical communities. Moreover, methods for the diagnosis and treatment of AR in China have not been standardized entirely and some patients are still treated according to regional preferences. Thus, the present guidelines have been developed by the Chinese Society of Allergy to be accessible to both national and international medical communities involved in the management of AR patients. These guidelines have been prepared in line with existing international guidelines to provide evidence-based recommendations for the diagnosis and management of AR in China.
Adult
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Asian Continental Ancestry Group*
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China
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Comorbidity
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Developed Countries
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Developing Countries
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Diagnosis*
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Epidemiologic Studies
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Epidemiology
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Global Health
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Humans
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Hypersensitivity*
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Prevalence
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Rhinitis, Allergic*
8.Analysis on gene sequence of HIV isolated from men who have sex with men in Beijing
Tao GUI ; Lei JIA ; Jingwan HAN ; Zuoyi BAO ; Siyang LIU ; Jingyun LI ; Lin LI
Chinese Journal of Epidemiology 2015;36(9):988-993
Objective To analyze genetic characteristics of HIV isolated from men who have sex with men(MSM) in Beijing and predict the epidemic trend in this population.Methods All of the HIV gene sequences in our laboratory obtained from MSM in Beijing were used,which were aligned with all of the HIV gene sequences from MSM and other populations in China downloaded from Los Alamos HIV Database.Phylogenetic trees were constructed by using software PhyML 3.0,based on which the relationships of prevalent HIV strains between Beijing MSM and other populations in China were further explored.The evolution rate,the time of most recent common ancestor (tMRCA),the epidemic parameters,the reproductive number (R0) were calculated by using software BEAST to predict HIV evolution and epidemic characteristics.Results Multiple HIV subtypes,including subtype B,CRF01_AE and CRF07_BC,were found to be prevalent among MSM in Beijing.In ML tree constructed based on strains from the whole country,three clusters including B-1,CRF01_AE-1,and CRF01_AE-2 were found among the MSM in Beijing (accounting for 40%).At least three independent introduction of B 1 cluster strains into Beijing MSM were found,which were at March 1991 (July 1984-February 1997),January 1994 (January 1989-January 1998),April 1991 (August 1984-January 1996).For CRF01_AE strains,two clusters including CRF01_AE-1 and CRF01_AE-2 were introduced into the population at December 2000 (March 1998-January 2003) and December 2001 (January 2000-July 2003) respectively.The population epidemiology of HIV in Beijing MSM was reconstructed based on sequences.The CRF01_AE-1 cluster spread more quickly than the other two clusters,and the evolution rate was higher.Conclusion Multiple HIV subtypes were found prevalent among MSM in Beijing.Although subtype B strain was introduced into Beijing MSM earlier than CRF01_AE strain,CRF01_AE strain increased more quickly than subtype B strain.More research and control of the CRF01_AE prevalence will be helpful for prevention and control of HIV epidemic in MSM in Beijing.
9.The evaluation a new kit for detecting whole range CRP
Yilong LI ; Meng WANG ; Zhiyi TANG ; Biag HAN ; Jingyun HAN ; Qian LIU
Chinese Journal of Laboratory Medicine 2009;32(9):1069-1073
CRP measurement on automated biochemistry analyzer. The method can be used in clinical diagnosis.
10.Posterior glass fiber-reinforced composite resin-bonded fixed partial dentures: A 3-dimensional modeling and finite element numerical analysis.
Jingyun HAN ; Renyuan FEI ; Yansheng LI ; Lei ZHANG
Journal of Biomedical Engineering 2006;23(4):756-761
The method of modeling and mesh generation about 3-unit tooth/restoration complex were established. The three-dimensional finite element models were subjected to four types of occlusal load applied to pontic element to evaluate 3 fiber framework designs and 3 cavities preparation configurations. By comparing the difference of stress distribution, following conclusions were obtained: the principal stress under buccal-lingual cusp load in traditional fiber framework pontic increased by 6.22% compared to that in pure composite resin pontic; optimized fiber framework obviously reduced stress level under any load; modified cavities exhibited better stress transfer and decreased share stress at adhesive interface than traditional cavities.
Composite Resins
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Dental Abutments
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Dental Materials
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Dental Stress Analysis
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Finite Element Analysis
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Glass
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Models, Theoretical

Result Analysis
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