1.Effects of Bushen Huoxue Formula (补肾活血方) on TLR/NF-κB Pathway and Intestinal Flora in Ileum Tissue of Parkinson's Disease Model Mice
Xiaorong QI ; Feiran HAO ; Xianglin TANG ; Fagen LI ; Yujia WANG ; Liang WANG ; Yingfan SHEN ; Minghui YANG ; Min LI
Journal of Traditional Chinese Medicine 2024;65(10):1038-1045
ObjectiveTo explore the possible mechanism of Bushen Huoxue Formula (补肾活血方, BHF) in the treatment of Parkinson's disease (PD) from the the perspective of intestinal flora. MethodsSeventy-two male C57/BL6J mice were randomly divided into blank group, model group, Madopar group and low-, medium- and high-dose BHF groups, with 12 mice in each group. The mice in the blank group were intraperitoneally injected with 10 ml/kg of normal saline, and those in the other groups were intraperitoneally injected with 30 mg/kg of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) at a concentration of 3 mg/ml to induce PD mice model, both once a day for 7 consecutive days. After successful modeling, the low-, medium-, and high-dose BHF groups were given 7.5, 15, and 30 g/(kg·d) of BHF by gavage, respectively, while the Madopar group was given 112.5 mg/(kg ·d) of Domedopar tablets by gavage, and the blank group and the model group were given 15 ml/(kg·d) of distilled water, all once a day for 14 consecutive days. The rod climbing test, rotating rod test, grip strength test and weight-bearing swimming test were used to evaluate the behavioral indicators of mice. Western blotting was used to measure the protein expression levels of Toll-like receptor (TLR)/nuclear factor kappa B (NF-κB) pathway inflammatory factors in the mouse ileum, including Toll-like receptor 2 (TLR2), Toll-like receptor 4 (TLR4), NF-κB, tumor necrosis factor α (TNF-α), interleukin 6 (IL-6), and interleukin 17 (IL- 17). 16S rRNA high-throughput sequencing was used to analyze changes in mouse intestinal flora. ResultsCompared to those in the blank group, the mice in the model group had longer bottoming time when climbing the pole, reduced grip strength, shortened rotary pole duration and swimming duration, and increased protein expression levels of TLR2, TLR4, NF-κB, TNF-α, and IL-6 in the ileal tissue (P<0.01). Compared to the model group, the Madopar group and the low-, medium- and high-dose BHF groups had shortened bottoming time of the climbing pole and increased grip strength; the Madopar group and the high-dose BHF group had prolonged rotary pole duration, and reduced protein expressions of TLR2, TLR4, NF-κB, TNF-α, IL-6, and IL-17 levels; and only the high-dose BHF group had prolonged swimming duration (P<0.05 or P<0.01). Compared to those in the low-dose BHF group, the bottoming time of the climbing pole were shorter in the moderate- and high-dose groups (P<0.05 or P<0.01), and the grip strength increased while the protein expression levels of TLR2, TLR4 and IL-17 decreased in the high-dose group (P<0.05 or P<0.01). The intestinal flora results showed significant differences between the blank group and the model group in the Dominance index, Pielou_e index, Shannon index, and Simpson index (P<0.05 or P<0.01). Compared to those of the model group, the Shannon index, Chao1 index, and Observed_otus index of the Madopar group, as well as the Chao1 index, Observed_otus index, Dominance index, Pielou_e index, Shannon index, and Simpson index of the high-dose BHF group all showed significantly statistical differences (P<0.05 or P<0.01). At the phylum level, the relative abundance categories of bacterial phyla with statistically significant differences in each group included Proteobacteria, Bacteroidetes, and Firmicutes (P<0.05 or P<0.01). At the genus level, the relative abundance categories of bacterial genera with statistically significant diffe-rences among each group included Muribaculaceae, Akkermansia, and Helicobacter pylori (P<0.05 or P<0.01). ConclusionThe possible mechanism of BHF in treating PD may be to reconstruct the disordered intestinal flora structure and improve the inflammatory response.
2.Status of anemia and its association with feeding patterns in infants and toddlers aged 6 - 24 months in Linxia Hui Autonomous Prefecture
Xiaolan MA ; Huiling WANG ; Ting CAO ; Xiaorong MA ; Juanjuan XU ; Qi HAN ; Haolan MA ; Xiaoyang HE ; Jianhua MA ; Gexiang ZHANG
Chinese Journal of Child Health Care 2024;32(3):273-279
【Objective】 To investigate the anemia status of infants and toddlers aged 6 - 24 months in Linxia Hui Autonomous Prefecture, Gansu Province, and to comprehensively evaluate the differences in feeding behaviors between anaemic and normal children through the infant and child feeding index (ICFI) and feeding knowledge scores, so as to provide reference for the guidance of infants and young children feeding in ethnic minority areas and the promotion of children′s growth and development. 【Methods】 Taking infants and young children aged 6 - 24 months in Linxia Prefecture as the study subjects, a multi-stage random sampling method was used to select children who met the requirements from 5 townships and 5 villages in 7 counties in 2019 and 2020.Periphral blood samples were collected to test the level of hemoglobin, so as to determine the anemia status.Meanwhile, physical examination was performed and a questionnaire survey of guardians was conducted to analyze the association betweenanaemia and feeding patterns 【Results】 A total of 3 901 infants and children were included in this study, of whom 729 (18.70%) were anaemic, with a mean ICFI score of 12.56±2.70 and a mean feeding knowledge score of 1.97±1.01.There was no statistically significant association of low feeding knowledge score and low ICFI with anaemia after adjusting for confounders (P>0.05), Unqualified meat addition in ICFI was a risk factor for anaemia (OR=1.355, P=0.042), while non-bottle feeding in the past 24 hours (OR=0.762, P=0.021), and breastfeeding in the past 24 hours of infants and toddlers aged 12 - 24 months (OR=0.228, P=0.018) were protective factor for anemia in infants and toddlers aged 12 - 24 months. 【Conclusions】 The average prevalence of anemia in infants and toddlers aged 6 - 24 months in Linxia Hui Autonomous Prefecture of Gansu Province is high, but the level of infant feeding and the level of feeding knowledge of caregivers are low.Early adherence to breastfeeding, timely addition of supplementary food, and more comsumpution of meat for children are conducive to preventing anemia.
3.Evaluation of disinfection effect of high-energy pulse ultraviolet disinfection equipment in medical institution settings
Zhengchun SUN ; Jincai ZHU ; Tingting ZHU ; Xiaorong QU ; Nan LI ; Yuxin ZHAO ; Shuang WANG ; Xiaodong YUAN ; Guoliang LI ; Nengwei ZHANG ; Yi XING ; Qi YAO
Chinese Journal of Preventive Medicine 2024;58(6):857-861
Objective:To evaluate the disinfection effect of high-energy pulse ultraviolet disinfection equipment in medical institution settings.Methods:The disinfection effect was evaluated through field tests and laboratory tests. Among them, 135 high-frequency contact points were selected from nine departments in the field test. Samples were collected before and after disinfection, and the disinfection effects of 75% alcohol wipes wiping disinfection, high-energy pulse ultraviolet disinfection robot disinfection and high-energy pulse ultraviolet handheld disinfection instrument were compared. In the laboratory test, 30 infected areas of the simulated test table were exposed to vertical ultraviolet irradiation and the bacterial-killing rate before and after disinfection was calculated.Results:In the field test, the bacteria-killing rates of 75% alcohol wipes, high-energy pulse ultraviolet disinfection robot and high-energy pulse ultraviolet handheld disinfection instrument were 94.99%, 91.53% and 95.94%, respectively, and the difference was statistically significant. The disinfection effect of the high-energy pulse ultraviolet handheld disinfection instrument was better than that of the high-energy pulse ultraviolet disinfection robot ( P values <0.05). In the laboratory test, the killing log value of Staphylococcus aureus and Escherichia coli on the carrier were both greater than 3.00. In the simulated field test, the killing log value of Staphylococcus aureus on the surface samples were 4.99. Conclusion:Both the high-energy pulse ultraviolet handheld disinfection instrument and the high-energy pulse ultraviolet disinfection robot have good disinfection effects, which are similar to the disinfection effects of conventional 75% alcohol wipes.
4.Evaluation of disinfection effect of high-energy pulse ultraviolet disinfection equipment in medical institution settings
Zhengchun SUN ; Jincai ZHU ; Tingting ZHU ; Xiaorong QU ; Nan LI ; Yuxin ZHAO ; Shuang WANG ; Xiaodong YUAN ; Guoliang LI ; Nengwei ZHANG ; Yi XING ; Qi YAO
Chinese Journal of Preventive Medicine 2024;58(6):857-861
Objective:To evaluate the disinfection effect of high-energy pulse ultraviolet disinfection equipment in medical institution settings.Methods:The disinfection effect was evaluated through field tests and laboratory tests. Among them, 135 high-frequency contact points were selected from nine departments in the field test. Samples were collected before and after disinfection, and the disinfection effects of 75% alcohol wipes wiping disinfection, high-energy pulse ultraviolet disinfection robot disinfection and high-energy pulse ultraviolet handheld disinfection instrument were compared. In the laboratory test, 30 infected areas of the simulated test table were exposed to vertical ultraviolet irradiation and the bacterial-killing rate before and after disinfection was calculated.Results:In the field test, the bacteria-killing rates of 75% alcohol wipes, high-energy pulse ultraviolet disinfection robot and high-energy pulse ultraviolet handheld disinfection instrument were 94.99%, 91.53% and 95.94%, respectively, and the difference was statistically significant. The disinfection effect of the high-energy pulse ultraviolet handheld disinfection instrument was better than that of the high-energy pulse ultraviolet disinfection robot ( P values <0.05). In the laboratory test, the killing log value of Staphylococcus aureus and Escherichia coli on the carrier were both greater than 3.00. In the simulated field test, the killing log value of Staphylococcus aureus on the surface samples were 4.99. Conclusion:Both the high-energy pulse ultraviolet handheld disinfection instrument and the high-energy pulse ultraviolet disinfection robot have good disinfection effects, which are similar to the disinfection effects of conventional 75% alcohol wipes.
5.Effects of moxibustion on CD4+T-cell proliferation by G6PD-mediated pentose phosphate pathway in adjuvant arthritis rats
Liang ZHANG ; Lingyun ZHAO ; Yiying LONG ; Jianan CAO ; Qirui QU ; Qingze WU ; Li LIU ; Xiaorong CHANG ; Kun AI ; Fang QI ; Yanping YANG
Journal of Acupuncture and Tuina Science 2024;22(4):273-279
Objective:To observe the effects of moxibustion on the levels of glucose-6-phosphate dehydrogenase(G6PD)and reduced nicotinamide adenine dinucleotide phosphate(NADPH)in the plasma and spleen and the CD4+T-cell number in the spleen of rats with adjuvant arthritis,thus to explore the mechanism in rheumatoid arthritis(RA)treatment with moxibustion by regulating the CD4+T-cell proliferation through G6PD-mediated pentose phosphate pathway. Methods:Twenty-seven male Sprague-Dawley rats were randomly divided into a blank group,a model group,and a moxibustion group,with 9 rats in each group.Incomplete Freund's adjuvant was used to induce inflammation in the model group and the moxibustion group.The blank group and the model group were not intervened.In the moxibustion group,suspended moxibustion was performed at bilateral Zusanli(ST36),Guanyuan(CV4),and Ashi points for 30 min,once a day for 24 times in total.Hematoxylin-eosin staining was used to evaluate the histopathological changes of rat synovial tissue;the swelling degree of the rat toes was observed by measuring the toe volume;G6PD and NADPH in the spleen and plasma were detected by Western blotting and enzyme-linked immunosorbent assay.Flow cytometry was used to detect the CD4+T-cell number in the spleen. Results:Compared with the blank group,the levels of G6PD and NADPH in the plasma and spleen and the CD4+T-cell number in the spleen were significantly increased in the model group(P<0.01 or P<0.05).Compared with the model group,the NADPH level in the spleen and plasma and the CD4+T-cell number in the spleen in the moxibustion group decreased significantly(P<0.05 or P<0.01),and the G6PD level in the plasma decreased significantly(P<0.05),but there was no significant difference in the G6PD level in the spleen(P>0.05). Conclusion:Moxibustion can regulate immunity and improve joint synovial inflammation in RA.The mechanism may be that the G6PD-mediated pentose phosphate pathway reduces the production of metabolite NAPDH in CD4+T cells,thereby inhibiting the proliferation of naive CD4+T cells.
6.Analysis of red blood cells supply before and after the outbreak of COVID-19 from 2018 to 2021 in 18 domestic blood centers
Dongyan ZHAO ; Hongwei MA ; Dingjie TANG ; Xiaorong FENG ; Hao TIAN ; Mengzhuo LUO ; Nan WU ; Yan LIN ; Xia DU ; Qi FU ; Junlei HUANG ; Changchun LU ; Xiaoli CAO ; Yi YANG ; Lin WANG ; Ying LI ; Hai QI ; Dongtai WANG ; Yan QIU
Chinese Journal of Blood Transfusion 2023;36(10):892-898
【Objective】 To compare the supply data of red blood cells(RBCs) from 18 blood centers in China before and after the outbreak of COVID-19 during 2018 to 2021. 【Methods】 Eight indicators related to RBCs supply from 18 blood centers in China during 2018-2021 were collected retrospectively, including the storage of total amount of qualified RBCs (referred to as the total amount of storage), the distribution of total amount of RBCs (referred to as the total amount of distribution), the distribution amount of RBCs per 1 000 population (referred to as the amount of distribution per 1 000 population), the distribution amount of RBCs from 400 mL original blood per 1 000 population [referred to as the amount of distribution per 1 000 population (400 mL)], the average daily distribution amount of RBCs (referred to as the average daily distribution amount), the average daily storage amount of RBCs (referred to as the average daily storage amount), the average storage days of RBCs when distribute (referred to as the RBC storage days), and the expired amount of RBCs (referred to as the expired amount). Based on the outbreak time of COVID-19, the data of 2018 and 2019 were the pre-pandemic group, and the data of 2020 and 2021 were the post-pandemic group. 【Results】 Data on RBCs supply in 18 blood centers from 2018 to 2021(comparison of the pre-pandemic group and the post-pandemic group): the amount of distribution per 1 000 population (median 14.68 U>13.92 U) decreased, the amount of distribution per 1 000 population (400 mL) (median 10.16 U>9.21 U) decreased, and the difference was statistically significant (P<0.05); data comparison between 2019 and 2020:the total amount of distribution (median 117 770.38 U>99 084.08 U) decreased, the amount of distribution per 1 000 population (median 15.04 U>12.19 U) decreased, the amount of distribution per 1000 population (400 mL) (median 10.11 U>8.94 U), the average daily distribution amount(322.66 U>270.73 U) decreased and RBC storage days (median 10.50 d<11.45 d) increased, the difference has statistical significance (P<0.05); data comparison between 2020 and 2021:the total amount of storage (median 101 920.25 U<120 328.63 U), the total amount of distribution (median 99 084.08 U<118 428.62 U), the amount of distribution per 1 000 population (median 12.19 U<15.00 U), the amount of distribution per 1 000 population (400 mL) (median 8.94 U<9.46 U), the average daily distribution amount (270.73 U>324.46 U), the average daily inventory (median 3 222.00 U<4 328.00 U) increased, the expired amount (median 1.50 U>0.00 U) decreased, the difference has statistical significance (P<0.05). The results of ANOVA showed that there were significant differences on the data related to RBCs supply (except expired amount) in different blood centers (P<0.05). The ratio of average daily stock to average daily distribution in the post-outbreak group (median 12.36 d) was higher than that in the pre-outbreak group (median 10.92 d), the difference has statistical significance (P<0.05), with significant difference among different blood centers (P <0.05). 【Conclusion】 The COVID-19 pandemic has a significant impact on RBCs supply in different blood centers. In the second year of the pandemic, the supply capability had recovered to some extent, and there were differences in RBCs supply in different blood centers.
7.Application of typical tasks-based mind mapping in nursing teaching for children with autism spectrum disorder
Jing CAO ; Jinhua HE ; Yingying HUANG ; Xiaojin CHEN ; Qi WANG ; Xiaorong LIU ; Jihong HU
Chinese Journal of Medical Education Research 2023;22(8):1276-1280
Objective:To explore the application of typical tasks-based mind mapping in nursing teaching for children with autism spectrum disorder (ASD).Methods:A total of 102 nursing students who were involved in the nursing of children with ASD in Hunan Children's Hospital were selected and divided into control group and observation group according to the teaching methods. Fifty-one students in the control group were provided with conventional teaching, while 51 students in the observation group were provided with typical tasks-based mind mapping teaching. The students in the two groups were assessed for performance, self-directed learning ability score, and overall literacy at completion of the nursing course. SPSS 22.0 was used for the t test. Results:The scores of theoretical examination[(92.34±4.07) vs. (89.92±3.61)], nursing note writing[(91.07±3.84) vs. (88.60±3.59)], and operational examination[(90.47±2.98) vs. (88.52±2.73)] were significantly higher among students in the observation group than among those in the control group ( P<0.05); after the internship, students in the two groups had significantly increased scores in interpersonal relationships, learning awareness, learning strategies, learning behaviors, and learning evaluation, and the observation group had better performance than the control group in the above indices ( P<0.05); after the internship, students in the two groups had significantly increased scores in problem solving, interpersonal communication, critical thinking, and self-leadership, and the observation group had better performance than the control group in the above indices ( P<0.05). Conclusion:The application of typical tasks-based mind mapping in nursing teaching for children with ASD can improve nursing students' academic performance, enhance their self-directed learning, and improve their overall literacy.
8.Intraoperative frozen pathology exam of Common iliac lymph nodes and Para-Aortic lymphadenectomy on the prognosis and quality of life for patients with IB2-IIA2 Cervical Cancer: trial protocol for a randomized controlled trial (C-PACC trial)
Xinyu QU ; Junjun QIU ; Lili JIANG ; Xiaorong QI ; Guonan ZHANG ; Weiwei FENG ; Yudong WANG ; Yincheng TENG ; Xipeng WANG ; Xiaoqing GUO ; Keqin HUA
Journal of Gynecologic Oncology 2023;34(2):e13-
Background:
The impact of para-aortic lymphadenectomy (PALD) on prognosis and quality of life (QoL) for IB2-IIA2 cervical cancer patients remain controversial. And whether intraoperative frozen pathology exam on common iliac lymph nodes could help predict para-aortic lymph node (PALN) metastasis was unanswered with high-level evidence.
Methods
A multi-center, randomized controlled study is intended to investigate the effect of PALD on the prognosis and QoL in cervical cancer patients and to assess the value of intraoperative frozen pathological evaluation of common iliac nodes metastasis for the prediction of PALN metastasis. After choosing whether to receive intraoperative frozen pathological examination of bilateral common iliac lymph nodes, eligible patients will be randomly assigned (1:1) to receive PALD or not. The primary end point is 2-year progression-free survival (PFS). The secondary end points include 5-year PFS, 2-year overall survival (OS), 5-year OS, adverse events (AEs) caused by PALD, AEs caused by radiotherapy and QoL. A total of 728 patients will be enrolled from 8 hospitals in China within 3-year period and followed up for 5 years.
9.miR-191 inhibits retinal vascular endothelial cell proliferation and angiogenesis
Qi ZHAO ; Weiting AN ; Boshi LIU ; Manhong XU ; Lijie DONG ; Xiaorong LI ; Jindong HAN
Chinese Journal of Ocular Fundus Diseases 2022;38(1):49-55
Objective:To observe the inhibitory effect of lentivirus (LV)-mediated miR-191 on the proliferation and angiogenesis of human retinal vascular endothelial cells (hREC) cultured in vitro.Methods:The hREC cell lines were cultured in vitro and divided into control group, hypoxia group, LV-empty vector (LV-vector) group, and LV-miR-191 (LV-191) group. The LV-vector group and LV-191 group were transferred to the corresponding lentiviral vector respectively. Flow cytometry was used to detect cell transfection efficiency. Cell Counting Kit-8 (CCK-8) test was used to detect cell proliferation ability. Scarification test and invasion chamber (Transwell) test were used to detect cell migration ability. Matrigel test was used to detect cell lumen formation ability. Real-time quantitative polymerase chain reaction (qPCR) was used to detect the relative expression of miR-191 and relative mRNA expression of its downstream target genes p21, vascular endothelial growth factor (VEGF), cell division protein kinase (CDK) 6, cyclin-D1 (Cyclin D1). Independent sample t test was used for pairwise comparison. Results:The results of flow cytometry showed that the transfection efficiency of cells in the control group and the LV-191 group were 0.615% and 99.400%, respectively. The results of CCK-8, scarification, Transwell and Matrigel test showed that, compared with the control group, the number of cell proliferation ( t=6.130, 4.606), the cell mobility ( t=4.910, 6.702), the number of stained cells on the microporous membrane ( t=7.244, 6.724) and the lumen formation ability cells ( t=8.345, 9.859) were significantly increased in the hypoxia group and the LV-vector group ( P<0.01), while the LV-191 group showed completely opposite performance ( t=14.710, 6.245, 5.333, 5.892; P≤0.01). The qPCR test results showed that, compared with the control group and the LV-vector group, the relative expression of miR-191 mRNA in the cells of the LV-191 group was significantly up-regulated ( t=44.110, 42.680), the relative expression of Cyclin D1 mRNA ( t=29.940, 14.010) and CDK6 mRNA ( t=15.200, 7.645) decreased significantly, and the difference were statistically significant ( P<0.01); the relative expression of p21 mRNA increased, however, the difference was not statistically significant ( t=2.013, 2.755; P>0.05). There was no significant difference in the relative expression of VEGF mRNA in the 4 groups of cells ( F=0.966, P>0.05). Conclusions:LV-191 can inhibit the proliferation, migration and tubing of hREC by up-regulating p21 and down-regulating CDK6 and Cyclin D1.
10.Enzyme-linked immunosorbent assays for quantification of MMMAE-conjugated ADCs and total antibodies in cynomolgus monkey sera
Pei MIN ; Liu TINGTING ; Ouyang LU ; Sun JIANHUA ; Deng XIAOJIE ; Sun XIAOMIN ; Wu WEI ; Huang PENG ; Chen YI-LI ; Tan XIAORONG ; Liu XIAOYUE ; Zhu PENG ; Liu YONGZHEN ; Wang DEHENG ; Wu JUNLIANG ; Wang QI ; Wang GUIFENG ; Gong LIKUN ; Qin QIUPING ; Wang CHUNHE
Journal of Pharmaceutical Analysis 2022;12(4):645-652
Antibody-drug conjugates(ADCs)are commonly heterogeneous and require extensive assessment of exposure-efficacy and exposure-safety relationships in preclinical and clinical studies.In this study,we report the generation of a monoclonal antibody against monomethyl auristatin E(MMAE)and the development,validation,and application of sensitive and high-throughput enzyme-linked immunosor-bent assays(ELISA)to measure the concentrations of MMAE-conjugated ADCs and total antibodies(tAb,antibodies in ADC plus unconjugated antibodies)in cynomolgus monkey sera.These assays were suc-cessfully applied to in vitro plasma stability and pharmacokinetic(PK)studies of SMADC001,an MMAE-conjugated ADC against trophoblast cell surface antigen 2(TROP-2).The plasma stability of SMADC001 was better than that of similar ADCs coupled with PEG4-Val-Cit,Lys(m-dPEG24)-Cit,and Val-Cit linkers.The developed ELISA methods for the calibration standards of ADC and tAb revealed a correlation be-tween serum concentrations and the OD450 values,with R2 at 1.000,and the dynamic range was 0.3-35.0 ng/mL and 0.2-22.0 ng/mL,respectively;the intra-and inter-assay accuracy bias%ranged from-12.2%to-5.2%,precision ranged from-12.4%to-1.4%,and the relative standard deviation(RSD)was less than 6.6%and 8.7%,respectively.The total error was less than 20.4%.The development and validation steps of these two assays met the acceptance criteria for all addressed validation parameters,which suggested that these can be applied to quantify MMAE-conjugated ADCs,as well as in PK studies.Furthermore,these assays can be easily adopted for development of other similar immunoassays.

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