1.Molecular biological research and molecular homologous modeling of Bw.03 subgroup
Li WANG ; Yongkui KONG ; Huifang JIN ; Xin LIU ; Ying XIE ; Xue LIU ; Yanli CHANG ; Yafang WANG ; Shumiao YANG ; Di ZHU ; Qiankun YANG
Chinese Journal of Blood Transfusion 2025;38(1):112-115
[Objective] To study the molecular biological mechanism for a case of ABO blood group B subtype, and perform three-dimensional modeling of the mutant enzyme. [Methods] The ABO phenotype was identified by the tube method and microcolumn gel method; the ABO gene of the proband was detected by sequence-specific primer polymerase chain reaction (PCR-SSP), and the exon 6 and 7 of the ABO gene were sequenced and analyzed. Homologous modeling of Bw.03 glycosyltransferase (GT) was carried out by Modeller and analyzed by PyMOL2.5.0 software. [Results] The weakening B antigen was detected in the proband sample by forward typing, and anti-B antibody was detected by reverse typing. PCR-SSP detection showed B, O gene, and the sequencing results showed c.721 C>T mutation in exon 7 of the B gene, resulting in p. Arg 241 Trp. Compared with the wild type, the structure of Bw.03GT was partially changed, and the intermolecular force analysis showed that the original three hydrogen bonds at 241 position disappeared. [Conclusion] Blood group molecular biology examination is helpful for the accurate identification of ambiguous blood group. Homologous modeling more intuitively shows the key site for the weakening of Bw.03 GT activity. The intermolecular force analysis can explain the root cause of enzyme activity weakening.
2.Effects of coal mine dust on lung function in rats
LIU Yang ; LI Meng ; LU Liyuan ; WANG Ru ; YANG He ; ZHANG Huifang
Journal of Preventive Medicine 2025;37(1):96-101
Objective:
To explore the impacts of coal mine dust on lung function in rats, so as to provide the basis for the early prevention and treatment of coal worker's pneumoconiosis.
Methods:
Seventy-two SPF-grade 8-week-old male Sprague-Dawley rats were randomly divided into the coal dust group, the coal-silica dust group, the silica dust group and the control group. The rats in the first three groups of rats were administered 1 mL corresponding dust suspension into the lungs using non-exposure tracheal instillation, while the rats in the control group were administered 1 mL normal saline. Respiratory rate (f), forced vital capacity (FVC), peak expiratory flow (PEF) and dynamic pulmonary compliance (Cdyn) were measured at 1, 3 and 6 months after dust exposure. Lung tissues were collected to measure reactive oxygen species (ROS) and adenosine triphosphate (ATP) levels using corresponding ELISA kits and ATP assay kits, respectively. The relative mRNA expressions of peroxisome proliferators-activated receptor gamma coactivator 1-alpha (PGC-1α) and mitochondrial transcription factor A (TFAM) were detected using real-time fluorescent quantitative polymerase chain reaction assay. The relative protein expressions of PGC-1α and TFAM were detected using Western blotting.
Results:
There was no interaction between dust type and exposure duration on f (P>0.05), but there were interactions on FVC, PEF and Cdyn (all P<0.05). Compared with the control group at 6 months after dust exposure, the f of the rats in the silica dust group were increased, while the FVC and PEF of the rats in the coal-silica dust and silica dust groups were decreased, and Cdyn of the rats in the coal dust, coal-silica dust and silica dust groups were decreased (all P<0.05). There were interactions between dust type and exposure duration on ROS and ATP levels, the relative mRNA and protein expressions of PGC-1α and TFAM (all P<0.05). Compared with the control group at 3 and 6 months after dust exposure, the ROS levels in the rats in the coal dust, coal-silica dust and silica dust groups were increased, while the ATP levels, the relative mRNA and protein expressions of PGC-1α and TFAM were decreased (all P<0.05).
Conclusion
The lung function impairment in rats caused by different types of coal mine dust is related to PGC-1α-mediated mitochondrial biogenesis dysfunction, which leads to increased ROS levels, decreased ATP and TFAM levels.
3.Dapagliflozin on acute kidney injury and prognosis in patients with diabetes mellitus type 2 and acute coronary syndrome after percutaneous coronary intervention
Huifang HAO ; Xiufeng ZHAI ; Qing LI ; Shicheng YANG ; Peng ZHANG
China Pharmacy 2025;36(4):469-474
OBJECTIVE To investigate the impact of dapagliflozin on contrast-induced acute kidney injury (CIAKI) and prognosis in patients with diabetes mellitus type 2 (T2DM) and acute coronary syndrome (ACS) who underwent percutaneous coronary intervention (PCI). METHODS Retrospective selection of data on T2DM patients with ACS who underwent PCI treatment in the Cardiology Department of Tianjin Chest Hospital from January 1st 2021 to December 31st 2022. The patients were divided into dapagliflozin group (96 cases) and control group (148 cases) based on whether they received dapagliflozin or not. Renal function indicators were measured for all enrolled patients before PCI and at 48 h and 1 week after PCI, including blood urea nitrogen (BUN), serum creatinine (Scr), estimated glomerular filtration rate (eGFR), cystatin-C (Cys-C), kidney injury molecule-1 (KIM-1) and β2-microglobulin (β2-MG). All patients were followed up for at least 1 year. The incidence of CIAKI and major adverse cardiac event (MACE) during follow-up were recorded for both groups. Logistic regression was used to analyze the impact of dapagliflozin on the occurrence of CIAKI, while the Log-rank test was applied to compare the incidence of MACE between the two groups. Cox regression was employed to analyze the impact of dapagliflozin on prognosis. RESULTS At 48 h and 1 week after PCI, serum levels of Cys-C, KIM-1 and β2-MG were significantly lower in the dapagliflozin group compared to the control group (P<0.05). The incidence of CIAKI was lower in the dapagliflozin group compared to the control group (6.25% vs. 14.86%, P=0.042). Logistic regression analysis revealed that dapagliflozin was an independent protective factor against CIAKI (OR=0.280, 95%CI 0.101-0.780,P=0.015). During the follow-up period, the incidence of MACE was lower in the dapagliflozin group compared to the control group (7.29% vs. 17.57%, P=0.049). Cox regression analysis indicated that dapagliflozin reduced the occurrence of MACE after PCI (HR=0.374, 95%CI 0.161-0.866, P=0.022). CONCLUSIONS With adequate hydration, the use of dapagliflozin does not increase the risk of CIAKI following PCI in T2DM patients with ACS.
4.Mechanism of immunosuppression in rats by herb-partitioned moxibustion based on transcriptome sequencing technology
Yuefeng TIAN ; Luojie XIONG ; Huifang WANG ; Chuntao ZHAI ; Wei LI
Chinese Journal of Tissue Engineering Research 2025;29(5):978-988
BACKGROUND:Immunosuppression leads to impaired body immune function and aggravates the disease.Herb-partitioned moxibustion can effectively regulate immune function and improve immunity in the body,but its regulatory mechanism has not been elucidated. OBJECTIVE:To sequence immunosuppressed model rats treated with herb-partitioned moxibustion using bioinformatics techniques based on transcriptomics and to explore the mechanisms by which it regulates immunity. METHODS:Twenty-four Sprague-Dawley rats were randomly assigned to three groups:control,model,and herb-partitioned moxibustion groups,with eight rats in each group.The model and herb-partitioned moxibustion groups were subjected to establishment of an immune suppression model by intraperitoneal injection of cyclophosphamide at a dose of 35 mg/kg for 3 consecutive days.No interventions were administered to the control and model groups after modeling.In contrast,the herb-partitioned moxibustion group received moxibustion treatment at Zhongwan,Shenque,Guanyuan,and Zusanli acupoints using a combination of moxa and herbal cakes,once a day,for 10 consecutive days,with samples being collected the day after the end of the intervention.Peripheral blood was collected from all groups of rats to measure their white blood cell count.RNA-seq was performed on the Illumina sequencing platform,and differentially expressed genes were selected for bioinformatics analysis using the GO and KEGG databases. RESULTS AND CONCLUSION:Compared with the control group,the model group exhibited a significant decrease in white blood cell count(P<0.001).RNA-seq analysis identified 3 026 differentially expressed genes between the model and control groups,with 1 565 upregulated and 1 461 downregulated.There were 535 differentially expressed genes identified between the herb-partitioned moxibustion group and the model group,with 280 upregulated and 255 downregulated.The Venn diagram analysis revealed that 159 genes were downregulated in the model group compared with the control group.However,after moxibustion with herbal cakes,these genes were upregulated.Protein-protein interaction network analysis identified 10 core targets,including Oasl,Oas2,Isg15,Herc6,Mx2,Helz2,Mx1,Syk,Hspa1a,and Ret.According to GO and KEGG analyses,moxibustion with herbal cakes regulated the body through pathways related to immune response,viruses,angiogenesis,and the autoimmune system.To conclude,there is a significant association between herbal cake-separated moxibustion intervention and immune suppression targets,including Oasl,Oas2,Isg15,Herc6,Mx2,Helz2,and Mx1.The intervention exhibits regulatory effects in the pathways related to immune responses,viral activities,and angiogenesis.
5.Effects of 3.0T magnetic resonance noise on the hearing of children aged 0-12 years with routine hearing protection
Huifang ZHAO ; Chao JIN ; Fangyao CHEN ; Cong TIAN ; Peiyao CHEN ; Xiaoyu WANG ; Miaomiao WANG ; Congcong LIU ; Xianjun LI ; Jian YANG
Journal of Xi'an Jiaotong University(Medical Sciences) 2024;45(1):145-149
Objective To explore the effect of exposure to noise of 3.0T magnetic resonance imaging(MRI)on children's cochlear function.Methods We prospectively recruited 72 children who underwent cranial MRI examination at our hospital from May to November 2018;3M earplugs and sponge mats were used for hearing protection during MRI scanning.Noise level(dBA)of each MRI sequence was detected with a nonmagnetic microphone and a sound level meter.Distortion product otoacoustic emissions(DPOAE)test at 2-7 kHz was performed 24 hours before and 30 minutes after the MRI examination.Paired t-test or Wilcoxon signed-rank test was used to analyze differences in DPOAE amplitude before and after the MRI examination.Results The average noise level of MRI measured in the study was(107.7±3.92)dBA.Compared with that before the MRI examination,the DPOAE amplitude(dB)changed little after the MRI examination;the range of amplitude differences in each age group was as follows:left ear(-0.24-1.10)and right ear(-0.24-0.74)in the 0-1 year-old group;left ear(-0.07-0.59)and right ear(-0.57-0.75)in the 2-5 year-old group;left ear(-0.36-0.44)and right ear(-0.30-0.57)in the 6-12 year-old group.No statistically significant difference was found(correction P>0.05).Conclusion No potential impact of 3.0T MRI noise on children's cochlear function was observed under routine hearing protection.
6.Prevalence and related factors of HIV testing among young students who ever had sexual experiences in Guangdong Province
Chinese Journal of School Health 2024;45(12):1718-1721
Objective:
To understand the prevalence of HIV testing and related factors among young students who had sex in Guangdong Province, in order to provide evidence for relevant education programs and HIV testing promotion in young students.
Methods:
From September to December 2022, a convenient sampling method was used to select 48 749 young students from 16 universities and mechanic colleges in 6 cities including Guangzhou, Shantou, Maoming, Huizhou, Dongguan, and Zhongshan in Guangdong Province for online questionnaire survey. A total of 2 971 students who ever had sexual experiences were screened out, and the HIV testing situation and related factors were investigated by using the questionnaire designed by AIDS Prevention and Education Project for College Students of China STD and AIDS Prevention Association.The influencing factors of HIV testing were analyzed using Chi square test and multiple Logistic regression model.
Results:
Among students who had sexual experiences, 11.92% (354/2 971) were tested for HIV. The results of multivariate Logistic regression analysis showed that among young sexual students, using psychoactive substances during sexual activity in the last 1 year ( OR =7.70), having first sex with the same sex ( OR =3.87), having commercial sex ( OR =2.37), having heard of PEP ( OR =2.20), having a high level of self assessed understanding of HIV testing ( OR =1.73), inconsistent use of condoms ( OR =1.56), being aware of HIV infection ( OR =1.53), being aware of HIV knowledge ( OR =1.51) were more likely to test for HIV, and females ( OR =0.39) were less likely to test for HIV ( P < 0.05).
Conclusions
The proportion of HIV testing is low among sexually active young students in Guangdong Province. Targeted interventions should be tailored to promote HIV testing coverage.
7.Influence and predictive value of systemic immune inflammation,fibrinogen and triglyceride glucose index on diabetic kidney disease
Chinese Journal of Diabetes 2024;32(3):197-202
Objective To investigate the influence and predictive value of systemic immune inflammation(SII),fibrinogen(Fib)and triglyceride glucose index(TyG)on the risk of diabetic kidney disease(DKD).Methods A total of 1073 patients with type 2 diabetes mellitus(T2DM)admitted to The Endocrinology Department of Hebei Yiling Hospital Affiliated to Hebei Medical University were enrolled in this study from January 2020 to December 2022.All the patients were divided into two groups according to the urinary protein creatinine ratio:DKD group(UACR≥30 mg/g,DKD,n=412)and simple T2DM group(UACR<30 mg/g,T2DM,n=661).Pearson correlation analysis was used to evaluat the correlation between SII,TyG,Fib and other indicators.Multiple logistic regression analysis was used to find the influencing factors for DKD.The clinical value of Fib,SII,TyG,and combined indicators in diagnosing DKD was tested by receiver operating characteristic(ROC)curve analysis.Results Compared with T2DM group,age,duration of DM,FPG,TG,TyG,cystatin C(CysC),glutamyl transpeptidase,SUA,homocysteine,SII,neutrophil count,Fib and D-D were increased(P<0.05 or P<0.01),while albumin(ALB)and HDL-C were decreased(P<0.05 or P<0.01)in DKD group.Pearson correlation analysis showed that SII was positively correlated with CysC,Scr,SUA,Fib,FDP,and D-D(P<0.05 or P<0.01),and negatively correlated with eGFR,ALB(P<0.01);TyG is positively correlated with SUA(P<0.01);Fib is positively correlated with CysC,Scr(P<0.01),and negatively correlated with eGFR(P<0.01).Multiple logistic regression analysis showed that TyG and Fib were the influencing factors for DKD.The ROC curve analysis showed that the area under the ROC curve for the combined index was 0.679,with a specificity of 74.2%,a sensitivity of 53.5%,and an optimal cutoff value of 0.276.Conclusion SII and Fib can directly cause renal function damage in DKD,while IR promotes DKD progression through metabolic disorders.The combined predictive value of Fib,SII,and TyG for DKD risk is superior to a single indicator.
8.Analysis for clinicopathological and immunohistochemical characteristics of patients with meibomian gland carcinoma
Man NIU ; Ying ZHAO ; Fengmei CAI ; Yuanpeng LI ; Wei QIAN ; Huifang WANG
International Eye Science 2024;24(11):1842-1845
AIM: To explore the clinicopathological and immunohistochemistry(IHC)characteristics of meibomian gland carcinoma(MGC).METHODS: Patients who were pathologically diagnosed as MGC from January 1, 2015 to December 31, 2020 in our hospital were enrolled, and their clinicopathological information was retrospectively analyzed. Cancer tissues from all the cases were IHC stained. En Vision two-step method, DAB staining, as well as hematoxylin re-staining were applied in the IHC assay.RESULTS: A total of 50 patients with 21 males and 29 females(1:1.38)were enrolled in the study, ranging from 26 to 80 years old, with a median age of 60 years. The upper eyelid, which was the predilection site, accounting for 66%(33/50). Histopathologically, moderately or poorly differentiated was in the majority(35/50, 70%). The expression rates of IHC parameters of MGC patients were as follows: GATA-3(49/50, 98%), EMA(49/50, 98%), CAM5.2(42/50, 84%), AR(41/50, 82%), MSH2(50/50, 100%), MSH6(50/50, 100%), MLH1(50/50, 100%), PMS2(50/50, 100%), Ki67(positive, 50%-90%). All the patients were followed up for 12 to 72 mo, with 5 cases of recurrence and 0 deaths.CONCLUSION: Pathological diagnosis of MGC should focus on observing cancer cells' cytoplasm to find relevant clues for cortical gland differentiation. Comprehensive analysis of multiple indicators is required when using IHC to assist diagnosis. For most MGC cancer cells, positive expressions of GATA-3, EMA, AR, CAM5.2 and a high Ki67 proliferation index could be always found. In addition, screening for Muir-Torre syndrome related IHC indicators could be also performed in diagnosing MGC simultaneously.
9.Involvement of let-7c in alveolar macrophage M2 polarization in coal mine dust-induced epithelial-mesenchymal transition in rat lung tissue
Ru WANG ; Liyuan LU ; Meng LI ; Yang LIU ; Xiaoyu HE ; Huifang ZHANG
Journal of Environmental and Occupational Medicine 2024;41(6):687-693
Background Coal workers' pneumoconiosis (CWP) is a serious occupational lung disease and one of the prescript occupational diseases in China. Epithelial-mesenchymal transition (EMT) is involved in the diffuse fibrosis of lung tissue of pneumoconiosis patients, and its mechanism may be related to the polarization of macrophages regulated by let-7c. Objective To investigate the effect of let-7c on the regulation of macrophage polarization in EMT in rats induced by coal dust exposure with different content of free SiO2. Methods SD rats were randomly divided into a control group, a 5% SiO2 group, a 30% SiO2 group, and a 99.9% SiO2 group, with 16 rats in each group. The rats in each group were tracheally titrated with 100 μL of 20 mg·mL−1 suspension (5% SiO2, 30% SiO2, and 99.9% SiO2) or normal saline, respectively. Alveolar lavage fluid was collected at the ends of the 1st month and the 3rd month. The relative expression levels of M1 or M2 markers, CD86 or CD206, in alveolar macrophages (AMs) were detected by immunofluorescence. The inflammation and fibrosis of lung tissue were observed by hematoxylin-eosin (HE) staining and Masson staining. The expression levels of transforming growth factor-β1 (TGF-β1), E-cadherin, and vimentin were detected by Western blotting. The expression levels of let-7c and c/EBP-δ genes were detected by real-time fluorescence quantitative PCR. Results The HE and Masson staining results showed that compared with the control group, the degree of pulmonary fibrosis in the 5% SiO2 group, the 30% SiO2 group, and the 99.9% SiO2 group gradually increased with the increase of dust exposure time. Compared with the control group, the expressions of CD86 and CD206 in the 5% SiO2 group, the 30% SiO2 group, and the 99.9% SiO2 group gradually increased at the end of the 1st month (F=330.904, 146.801, P<0.05), and the expression of CD86 in each group decreased gradually at the end of the 3rd month (F=331.781, P<0.05), but the expression of CD206 increased (F=1164.190, P<0.05). At the end of the 1st month, the expressions of TGF-β1 (F=8.847, P<0.05) and vimentin (F=13.275, P<0.05) gradually increased, and the expression of E-cadherin (F=6.253, P<0.05) gradually reduced in the 5% SiO2 group, the 30% SiO2 group, and the 99.9% SiO2 group. At the end of the 3rd month, the expressions of TGF-β1 (F=16.833, P<0.05) and vimentin (F=55.021, P<0.05) increased, and the expression of E-cadherin (F=12.790, P<0.05) gradually decreased in all groups. The PCR results showed that compared with the control group, the expression of let-7c mRNA in the 5% SiO2 group, the 30% SiO2 group, and the 99.9% SiO2 group increased at the ends of the 1st month and the 3rd month (F=11.251, 28.136, P<0.05). The expression of c/EBP-δ mRNA decreased in all groups at the ends of the 1st month and the 3rd month (F=49.204, 177.090, P<0.05). Conclusion In response to dust stimulation, let-7c promotes EMT by modulating macrophage polarization, which is involved in the formation of pulmonary fibrosis and thus influences the progression of CWP .
10.Effects of coal mine dust on interleukin-6 and let-7e in rats
LI Baichun ; SUN Yuhan ; ZHANG Huifang ; LU Xiaoting ; SONG Jing ; KONG Xiaomei ; WANG Linping
Journal of Preventive Medicine 2024;36(2):93-96
Objective:
To investigate the changes in the levels of interleukin-6 (IL-6) and let-7e in rats induced by coal mine dust, so as to provide the basis for the mechanism of coal worker's pneumoconiosis (CWP).
Methods:
Sixty-four clean and healthy male Sprague-Dawley rats were randomly divided into the control group, coal dust group, mixed dust group (mixed coal and silica dust) and quartz group. The rats in the control group were exposed to 1 mL physiological saline by non-exposure tracheal perfusion, and the rats in the dust-exposed groups were exposed to 1 mL dust suspension. Rats were sacrificed by anesthesia after 1 month and 6 months, lung tissue was observed using hematoxylin-eosin staining, the pathological change in the lungs was scored using the Szapiel scoring system, the levels of IL-6 in the bronchoalveolar lavage fluid were detected using enzyme-linked immunosorbent assay, and the expression of let-7e was determined by quantitative real-time PCR.
Results:
A month after exposure, a small amount of coal spots and inflammatory exudation were observed in the lung tissue of the coal dust group and the mixed dust group. The quartz group showed tissue structure destruction and mild fibrosis and thickening of alveolar septum. Six months after exposure, there were more coal spots and slightly thickened alveolar septum in the coal dust group, and hyperplasia of pulmonary interstitial fibers, destruction of alveolar structure and silica nodules were observed in the mixed dust group. In the quartz group, the alveolar structure was obviously destroyed, the interstitial fiber proliferation was significant and silica nodules were seen. Two-factor analysis of variance showed that the interaction between duration of exposure and dust type significantly influenced the pathological score of lung tissue, IL-6 levels, and let-7e expression levels (P<0.05). Under the same dust type, the pathological score of lung tissue and IL-6 levels were higher at 6 months after exposure than at 1 month, while the relative expression of let-7e was lower at 6 months after exposure than at 1 month (all P<0.05). Under the same duration of exposure, the pathological score of lung tissue and IL-6 levels were higher in the dust-exposed groups than in the control group, while the relative expression of let-7e was lower in the dust-exposed groups than in the control group (all P<0.05).
Conclusions
Coal dust can cause an increase in levels of IL-6 and a decrease in let-7e expression in rats. The type of dust and duration of exposure can interactively affect IL-6 and let-7e.


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