1.Quantitative Analysis of Multi-components in Gnaphalium affine Based on UPLC-Q-Exactive Orbitrap MS
Chunyan CAO ; Jiaqi ZHANG ; Zhenduo ZHAO ; Qin SHEN ; Shenglan QI ; Wei LIU ; Lichao ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(9):148-155
ObjectiveTo establish a rapid and stable liquid chromatography-mass spectrometry(LC-MS) for simultaneous analysis of 17 chemical components in Gnaphalium affine aboveground parts with flowers, so as to provide experimental basis for improving the quality standard of this herb. MethodUltra performance liquid chromatography-quadrupole/electrostatic field orbitrap mass spectrometry(UPLC-Q-Exactive Orbitrap MS) was used for the quantitative analysis of 17 constituents in 15 batches of G. affine from different origins, the separation was performed on an ACQUITY UPLC® BEH C18 column(2.1 mm×100 mm, 1.7 μm) with the mobile phase of methanol(A)-0.1% formic acid aqueous solution(B) for gradient elution(0-1.0 min, 8%A; 1.0-4.0 min, 8%-26%A; 4.0-9.0 min, 26%A; 9.0-14.0 min, 26%-34%A; 14.0-14.5 min, 34%-45%A; 14.5-15.0 min, 45%-60%A; 15.0-18.0 min, 60%-90%A; 18.0-19.0 min, 90%A; 19.0-19.01 min, 90%-8%A; 19.01-20.0 min, 8%A), the flow rate was 0.3 mL·min-1, the column temperature was 40 ℃ and the injection volume was 2 μL. And the electrospray ionization was used with full scanning in both positive and negative ion modes, and the scanning range was m/z 100-1 000. ResultThe established method has been verified by the methodology and could be used for the simultaneous quantification of 17 components in G. affine. The content ranges of the 17 components(quinic acid, gallic acid, protocatechuic acid, neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, 1,3-O-dicaffeoylquinic acid, isochlorogenic acid A, isoquercitrin, 1,5-O-dicaffeoylquinic acid, apigenin-7-O-glucoside, astragalin, isochlorogenic acid C, luteolin, apigenin and hispidulin) in 15 batches of G. affine samples was 39.60-179.12, 0.17-0.84, 2.41-8.38, 4.33-31.50, 13.63-180.38, 2.43-14.75, 1.16-19.68, 0.49-5.63, 55.77-445.16, 0.23-10.26, 62.04-530.10, 1.11-18.01, 11.36-90.61, 12.22-65.98, 7.22-69.84, 3.37-45.65, 0.30-2.59 μg·g-1, respectively. The content of organic acids was higher than that of flavonoids in G. affine, and the contents of 1,5-O-dicaffeoylquinic acid, isochlorogenic acid A, quinic acid and chlorogenic acid were higher. Meanwhile, the content of flavonoids in the samples from Guizhou was higher than that from Jiangsu, while the content of organic acids in the samples from Jiangsu was higher than that from Guizhou. ConclusionThe established method can be used for the rapid and accurate determination of 17 components in G. affine, which clarifies the content range of the main components in this herb, and can provide a reference for the selection of quality control markers of G. affine.
2.Evaluation of the safety and efficacy of mitomycin C-perfluorooctyl bromide liposome nanoparticles in the treatment of human pterygium fibroblasts
Tao LI ; Lingshan LIAO ; Shenglan ZHU ; Juan TANG ; Xiaoli WU ; Qilin FANG ; Ying LI ; Biao LI ; Qin TIAN ; Junmei WAN ; Yi YANG ; Yueyue TAN ; Jiaqian LI ; Juan DU ; Yan ZHOU ; Dan ZHANG ; Xingde LIU
Recent Advances in Ophthalmology 2024;44(2):100-105
Objective To prepare a nano drug(PFOB@Lip-MMC)with liposome as the carrier,liquid perfluorooc-tyl bromide(PFOB)as core and mitomycin C(MMC)loading on the liposome shell and study its inhibitory effect on the proliferation of human pterygium fibroblasts(HPFs).Methods The thin film dispersion-hydration ultrasonic method was used to prepare PFOB@Lip-MMC and detect its physical and chemical properties.Cell Counting Kit-8,Cam-PI cell viability staining and flow cytometry were employed to detect the impact of different concentrations of PFOB@Lip-MMC on the via-bility of HPFs.DiI fluorescence labeled PFOB@Lip-MMC was used to observe the permeability of the nano drug to HPFs under a laser confocal microscope.After establishing HPF inflammatory cell models,they were divided into the control group(with sterile phosphate-buffered saline solution added),PFOB@Lip group(with PFOB@Lip added),MMC group(with MMC added),PFOB@Lip-MMC group(with PFOB@Lip-MMC added)and normal group(with fresh culture medi-um added)according to the experimental requirements.After co-incubation for 24 h,flow cytometer was used to detect the apoptosis rate of inflammatory cells,and the gene expression levels of interleukin(IL)-1β,prostaglandin E2(PGE2),tumor necrosis factor(TNF)-α and vascular endothelial growth factor(VEGF)in cells were analyzed by PCR.Results The average particle size and Zeta potential of PFOB@Lip-MMC were(103.45±2.17)nm and(27.34±1.03)mV,respec-tively,and its entrapped efficiency and drug loading rate were(72.85±3.28)%and(34.27±2.04)%,respectively.The sustained-release MMC of drug-loaded nanospheres reached(78.34±2.92)%in vitro in a 24-hour ocular surface environ-ment.The biological safety of PFOB@Lip-MMC significantly improved compared to MMC.In terms of the DiI fluorescence labeled PFOB@Lip-MMC,after co-incubation with inflammatory HPFs for 2 h,DiI fluorescence labeling was diffusely dis-tributed in the cytoplasm of inflammatory HPFs.The apoptosis rate of inflammatory HPFs in the PFOB@Lip-MMC group[(77.23±4.93)%]was significantly higher than that in the MMC group[(51.62±3.28)%].The PCR examination results showed that the gene transcription levels of IL-1 β,PGE2,TNF-α and VEGF in other groups were significantly reduced com-pared to the control group and PFOB@Lip group,with the most significant decrease in the PFOB@Lip-MMC group(all P<0.05).Conclusion In this study,a novel nano drug(PFOB@LIP-MMC)that inhibited the proliferation of HPFs was successfully synthesized,and its cytotoxicity was significantly reduced compared to the original drugs.It has good bio-compatibility and anti-inflammatory effects,providing a new treatment approach for reducing the recurrence rate after pte-rygium surgery.
3.Application of OSCE-guided Scenario-based Practical Teaching Model in Clinical Pharmacist Training
Tiantian TANG ; Yiwen XIAO ; Haiyan YUAN ; Qiong LU ; Ying WANG ; Wenhui LIU ; Shenglan TAN ; Bikui ZHANG ; Daxiong XIANG ; Yan CHENG ; Yangang ZHOU ; Ping XU
Chinese Journal of Modern Applied Pharmacy 2024;41(10):1409-1414
OBJECTIVE
To explore the specific application and evaluation effect of objective structured clinical examination(OSCE)-guided scenario-based practical teaching mode in training clinical pharmacists.
METHODS
Fifty-six trainees who participated in the clinical pharmacist training program in the Second Xiangya Hospital of Central South University from October 2020 to September 2022 were selected as the research objects. OSCE-guided teaching was conducted, and the application effect of OSCE-guided teaching mode in clinical pharmacist training was explored and analyzed by using theoretical examination results and OSCE assessment results as evaluation indicators.
RESULTS
Through comparative analysis, it was found that the OSCE-guided teaching mode not only enabled students to better grasp the theoretical knowledge points required by the training outline, but also improved their clinical thinking ability, problem-solving ability, and communication and coordination skills to varying degrees.
CONCLUSION
For clinical pharmacist trainees, the OSCE teaching mode is conducive to the comprehensive improvement of clinical pharmacist skills and is suitable for cultivating clinical pharmacists who are capable of independently carrying out clinical pharmacy services in the new situation.
4.Feasibility of preparing human plasminogen by full chromatography from precipitation of Fraction Ⅲ in low temperature ethanol method
Jin ZHANG ; Shenglan YUE ; Chen ZHU ; Yan PENG ; Yanxiang ZHOU ; Lianzhen LIN ; Kejin CHEN ; Lu FENG ; Yong HU ; Zhijun ZHOU
Chinese Journal of Blood Transfusion 2024;37(11):1293-1300
[Objective] To determine the feasibility of preparing plasminogen (Pg) with Fraction Ⅲ precipitation (hereinafter referred to as FⅢ-P) from low-temperature ethanol process by full chromatography (hereinafter referred to as FⅢ-P process). [Methods] The FⅢ-P was diluted with dissolution buffer at different dilution times and stirring time. The potency and antigen concentration of Pg in dissolution sample were detected and the dissolution and clarification conditions were determined. Pre-treatment of loading sample and pre-experiment of affinity chromatography were carried out on the FⅢ-P dissolution sample to judge whether the loading sample had an impact on the chromatography by observing the performance of the affinity chromatography column and to evaluate whether the affinity chromatography could achieve the purpose of purifying Pg by detecting the plasma protein antigen concentration and Pg potency of the samples in the process. Two batches of FⅢ-P process were studied step by step, and the specific activity, steps and total recovery, and the output of Pg per ton of plasma were calculated. The feasibility of preparing Pg by FⅢ-P process was evaluated by comparing with the data of full chromatography process using plasma as raw material (hereinafter referred to as plasma process). [Results] The FⅢ-P was dissolved with 10 times of dissolution buffer, stirred for 1 hour, centrifuged at room temperature of 10 000×g for 15 minutes. The supernatant was first filtered with a screen, then clarified with an 8/0.8 μm filter, and finally filtered with a 0.45/0.2 μm filter and loaded. Pre-test showed that from clarification and filtration to Pg affinity chromatography, the step recovery of activity and antigen was 39.51% and 108.64%, respectively, the antigen concentration of Pg increased by 31.16 times and the activity increased by 11.39 times after affinity chromatography, which reaching the effect of affinity chromatography purification of Pg. The results of 2 batches of step-by-step scale-up FⅢ-P process showed that the total recoveries of antigen and activity from plasma to SP chromatography of FⅢ-P process were (45.76±1.10)% and (24.15±0.59)%, respectively, which had a total loss of about 1/3 of antigen and about 2/3 of activity compared to the plasma process. The Pg specific activity of SP chromatography eluent was (4.68±0.25) U/mg, which was about half of that of plasma process, but meeted the internal standard of > 4 U/mg. The output of Pg antigen per ton of plasma in the FⅢ-P process was 68.73% of that in the plasma process, and the output of Pg activity per ton of plasma in the plasma process was 29.82% of that in the plasma process, which basically achieved the purpose of waste utilization of FⅢ-P. [Conclusion] The technical route of preparing Pg from FⅢ-P by full chromatography is feasible.
5.Current status of missed nursing care during labor and its influencing factors
Shenglan DING ; Yilan ZHANG ; Qingxia WANG ; Xiujuan FU ; Xin WANG
Chinese Journal of Practical Nursing 2024;40(34):2673-2681
Objective:To investigate the current state of missed nursing care during labor, explore the reasons why nursing care was missed, and examine the influencing factors of missed nursing care.Methods:Data were collected from 14 medical institutions of different levels in southwest China using a convenience sampling from February to April, 2023. A total of 491 midwives were included. A cross-sectional survey was conducted using a general information questionnaire and the Chinese version of the Perinatal Missed Care. Multiple linear regression analysis was conducted to identify the influencing factors on missed nursing care.Results:Among 491 midwives, there are 6 males and 485 females. 194 midwives aged ≤ 30, 205 midwives aged 31-40 years, and 92 midwives aged ≥41 years. 80.45%(395/491) of midwives reported missing at least one item of nursing care during labor in the past week, and the missing rate of single item ranged from 13.24% (65/491) to 53.36% (262/491). The average score across 23 items of missed nursing care during labor was 1.49 ± 0.58. The average scores of items spanning the three dimensions of basic care, necessary care and postnatal care were 1.67 ± 0.68, 1.36 ± 0.56, and 1.51 ± 0.70, respectively. The average score of 13 items representing reasons for missed nursing care was 1.82 ± 0.71. The average scores of items in three dimensions of communication, labor resources, and material resources were 1.75 ± 0.73, 1.97 ± 0.87, and 1.74 ± 0.83, respectively. The multiple linear regression analyses showed that gender, monthly average income level, and whether overtime hours were included in total working hours were independently associated with missed nursing care (all P<0.05). Conclusions:The current state of missed nursing care during labor needs to be improved. Missed basic care is common, and labor resources are the significant reasons, and is influenced by factors such as demographic and sociological factors. Human resource factors are an important reason for nursing deficiency, and targeted measures should be taken to reduce the occurrence of nursing deficiency.
6.Exploring the causal association between testosterone and atherosclerosis based on a two-sample Mendelian randomization analysis
Jiayi HU ; Hongcen LIU ; Shenglan GONG ; Tian XIE ; Tianfeng ZHANG
Chinese Journal of Arteriosclerosis 2024;32(4):339-346
Aim To study whether genetically predicted serum testosterone level is causally associated with sys-temic multisite atherosclerosis.Methods Based on two pooled databases of genome-wide association studies on testos-terone and atherosclerosis in European populations from two separate foreign countries,the causal effect between testosterone and atherosclerosis was assessed using two-sample Mendelian randomization analysis with data on testosterone-associated genetic variants as instrumental variables(Ⅳ),and by using the inverse-variance weighted(IVW)method,MR-Egger regression,and weighted median estimation.Results IVW results showed that genetically predicted circu-lating testosterone levels were negatively associated with the risk of peripheral atherosclerosis(OR=0.93,95%CI:0.86~1.00,P=0.01),and that elevated testosterone level may reduce the risk of developing peripheral atherosclerosis,while no evidence of a potential causal association was found with cerebral atherosclerosis,coronary atherosclerosis and other athero-sclerosis type(P>O.05).Conclusion The final analysis showed a causal relationship between genetically predicted testosterone level and the risk of developing peripheral atherosclerosis,and the role of testosterone therapy in the prevention and treatment of atherosclerosis deserves attention and further study.
7.The effects of occupational noise exposure on glycated hemoglobin and prediabetes
Xiayun DAI ; Wenjun YIN ; Jichao LI ; Ling ZHANG ; Tian XU ; Xiaoli SHEN ; Qunyan LI ; Shenglan ZHOU ; Guilin YI ; Zhiwei PAN ; Zhenlong CHEN
China Occupational Medicine 2023;50(3):274-278
Objective To investigate the relationship between occupational noise exposure and glycated hemoglobin (HbA1c) levels, as well as prediabetes diagnosed by HbA1c. Methods A total of 1 181 workers from a cigarette factory were selected as the research subjects using a judgment sampling method. Workers were divided into control, low-level noise exposure and high-level noise exposure groups, consisting of 236, 359, and 586 individuals, respectively. The blood sample was collected for HbA1c test and occupation noise exposure intensity in workplace was detected by an area-sampling method. Results There were no statistical significant differences in HbA1c levels and prediabetes prevalence among the three groups of workers (all P>0.05). After adjusting for potential confounding factors such as years of service, gender, smoking, pack-years of smoking, alcohol consumption, and body mass index, multiple linear regression analysis showed that the high-level noise exposure group had higher HbA1c level than the control group (P<0.05). Multivariable logistic regression analysis results showed that the high-level noise exposure group had higher risk of prediabetes compared with the control group (P<0.05). Conclusion Occupational noise exposure could be a risk factor for the increased HbA1c levels and prediabetes incidence among the occupational population. More attention should be paid to the effects of occupational noise exposure on the HbA1c level in occupational health surveillance.
8.Role of melatonin receptor 1B gene polymorphism and its effect on the regulation of glucose transport in gestational diabetes mellitus.
Lijie WEI ; Yi JIANG ; Peng GAO ; Jingyi ZHANG ; Xuan ZHOU ; Shenglan ZHU ; Yuting CHEN ; Huiting ZHANG ; Yuanyuan DU ; Chenyun FANG ; Jiaqi LI ; Xuan GAO ; Mengzhou HE ; Shaoshuai WANG ; Ling FENG ; Jun YU
Journal of Zhejiang University. Science. B 2023;24(1):78-88
Melatonin receptor 1B (MT2, encoded by the MTNR1B gene), a high-affinity receptor for melatonin, is associated with glucose homeostasis including glucose uptake and transport. The rs10830963 variant in the MTNR1B gene is linked to glucose metabolism disorders including gestational diabetes mellitus (GDM); however, the relationship between MT2-mediated melatonin signaling and a high birth weight of GDM infants from maternal glucose abnormality remains poorly understood. This article aims to investigate the relationship between rs10830963 variants and GDM development, as well as the effects of MT2 receptor on glucose uptake and transport in trophoblasts. TaqMan-MGB (minor groove binder) probe quantitative real-time polymerase chain reaction (qPCR) assays were used for rs10930963 genotyping. MT2 expression in the placenta of GDM and normal pregnant women was detected by immunofluorescence, western blot, and qPCR. The relationship between MT2 and glucose transporters (GLUTs) or peroxisome proliferator-activated receptor γ (PPARγ) was established by western blot, and glucose consumption of trophoblasts was measured by a glucose assay kit. The results showed that the genotype and allele frequencies of rs10830963 were significantly different between GDM and normal pregnant women (P<0.05). The fasting, 1-h and 2-h plasma glucose levels of G-allele carriers were significantly higher than those of C-allele carriers (P<0.05). Besides, the protein and messenger RNA (mRNA) expression of MT2 in the placenta of GDM was significantly higher than that of normal pregnant women (P<0.05). Melatonin could stimulate glucose uptake and GLUT4 and PPARγ protein expression in trophoblasts, which could be attenuated by MT2 receptor knockdown. In conclusion, the rs10830963 variant was associated with an increased risk of GDM. The MT2 receptor is essential for melatonin to raise glucose uptake and transport, which may be mediated by PPARγ.
Female
;
Humans
;
Pregnancy
;
Blood Glucose/metabolism*
;
Diabetes, Gestational/metabolism*
;
Glucose/metabolism*
;
Melatonin/metabolism*
;
Polymorphism, Genetic
;
PPAR gamma
;
Receptor, Melatonin, MT2/genetics*
9.Clustering of health-risk behaviors among children and adolescents in Yancheng City
LU Xiaomin ; ZHAO Jijuan ; SUN Zhongyou ; ZHANG Xueyan ; CHEN Shenglan
Journal of Preventive Medicine 2023;35(4):331-334
Objective:
To investigate the clustering of health-risk behaviors and its influencing factors among children and adolescents in Yancheng City, Jiangsu Province, so as to provide insights into the prevention and control of health-risk behaviors among children and adolescents.
Methods:
Students were randomly sampled from 4 primary schools, 4 junior high schools and 4 senior high schools in Yancheng City using a multi-stage stratified cluster random sampling method from September to December 2021. Students' demographics and 12 health-risk factors including unhealthy diet, insufficient physical activity and attempted smoking were collected using the Student's Health Status and Influencing Factors Questionnaire by Jiangsu Provincial Center for Disease Control and Prevention, and factors affecting the clustering of health-risk behaviors were identified using a multivariable linear regression model.
Results:
A total of 2 925 valid questionnaires were recovered, and the respondents included 1 611 boys (55.08%) and 1 314 girls (44.92%). A total of 2 896 respondents were detected with health-risk behaviors, with a detection rate of 99.09%, and 2 772 respondents were detected with clustering of health-risk behaviors (93.06%). Insufficient sleep, insufficient physical activity and insufficient duration of outdoor activity were predominant patterns of clustering. The median number of health-risk behaviors was 4.00 (interquartile range, 2.00) per capita. Multivariable linear regression analysis showed that boys (β=0.232), grade (junior high school, β=0.519; senior high school, β=0.427), urban area (β=0.241), living at school (β=0.395), family structure (single parental family, β=0.188; other families, β=0.344) and father's education level of primary school and below (β=0.369) were factors affecting clustering of health-risk behavior among primary and high school students.
Conclusions
The detection of health-risk behaviors is high among children and adolescents in Yancheng City, and insufficient sleep, insufficient physical activity and insufficient duration of outdoor activity are predominant health-risk behaviors. Boys, junior high school and above, urban areas, living at schools, single parents, and fathers with a low educational level lead to a high degree of clustering of health-risk behaviors.
10.Clinical significance of PDGFRβ gene testing in hematological tumors.
Mengqiao GUO ; Fangyu GUO ; Yan ZHANG ; Hui CHENG ; Gusheng TANG ; Zhengxia HUANG ; Shenglan GONG
Chinese Journal of Medical Genetics 2023;40(11):1334-1339
OBJECTIVE:
To explore the clinical and laboratory characteristics of hematological tumors with different types of abnormalities in platelet derived growth factor β (PDGFRβ) gene.
METHODS:
A retrospective analysis was carried out on 141 patients with abnormal long arm of chromosome 5 (5q) and comprehensive medical history data from Changhai Hospital Affiliated to Naval Medical University from 2009 to 2020, and their clinical data were collected. R-banding technique was used for chromosomal karyotyping analysis for the patient's bone marrow, and fluorescence in situ hybridization (FISH) was used to detect the PDGFRβ gene. The results of detection were divided into the amplification group, deletion group, and translocation group based on FISH signals. The three sets of data column crosstabs were statistically analyzed, and if the sample size was n >= 40 and the expected frequency T for each cell was >= 5, a Pearson test was used to compare the three groups of data. If N < 40 and any of the expected frequency T for each cell was < 5, a Fisher's exact test is used. Should there be a difference in the comparison results between the three sets of data, a Bonferroni method was further used to compare the data.
RESULTS:
In total 98 patients were detected to have PDGFRβ gene abnormalities with the PDGFRβ probe, which yielded a detection rate of 69.50% (98/141). Among these, 38 cases (38.78%) had PDGFRβ gene amplifications, 57 cases (58.16%) had deletions, and 3 (3.06%) had translocations. Among the 98 cases, 93 were found to have complex karyotypes, including 37 cases from the amplification group (97.37%, 37/38), 55 cases from the deletion group (96.49%, 55/57), and 1 case from the translocation group (33.33%, 1/3). Analysis of three sets of clinical data showed no significant gender preponderance in the groups (P > 0.05). The PDGFRβ deletion group was mainly associated with myeloid tumors, such as acute myeloid leukemia (AML) and myelodysplastic syndrome (MDS) (P < 0.001). The PDGFRβ amplification group was more common in lymphoid tumors, such as multiple myeloma (MM) (P < 0.001). The PDGFRβ translocation group was also more common in myelodysplastic/myeloproliferative tumors (MDS/MPN).
CONCLUSION
Tumors with PDGFRβ gene rearrangement may exhibit excessive proliferation of myeloproliferative tumors (MPN) and pathological hematopoietic changes in the MDS, and have typical clinical and hematological characteristics. As a relatively rare type of hematological tumor, in addition to previously described myeloid tumors such as MPN or MDS/MPN, it may also cover lymphoid/plasma cell tumors such as multiple myeloma and non-Hodgkin's lymphoma.
Humans
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Clinical Relevance
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Hematologic Neoplasms/genetics*
;
In Situ Hybridization, Fluorescence
;
Multiple Myeloma
;
Myelodysplastic Syndromes
;
Retrospective Studies
;
Translocation, Genetic


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