1.Establishment of a method for detecting the potency of recombinant human coagulation factor Ⅶa for injection
Rong WU ; Liping WANG ; Jinye LANG ; Yue ZHU ; Jing ZHOU ; Xun LIU ; Jing NI ; Shunbo ZHOU ; Yaling DING
Chinese Journal of Blood Transfusion 2025;38(3):415-420
[Objective] To establish a method for detecting the potency of recombinant human coagulation factor Ⅶa for injection. [Methods] By adding the sample and factor Ⅶ deficient plasma to the sample cup and activating the reaction with prothrombin time assay reagent (PT reagent), the coagulation time of the sample was determined by the change in magnetic bead swing amplitude in the sample cup. The logarithm of coagulation time was inversely proportional to the logarithm of human factor Ⅶa potency. [Results] Under the experimental conditions, the specificity of the methodology was evaluated through spiked recovery, and the recovery rates ranged from 90.0% to 110.0%. Within the range from 0.125 to 1.000 IU/mL, there was a good linear response between the potency and coagulation time of the standard and sample, with correlation coefficients r>0.99. As for the accuracy and repeatability, the recovery rates of various concentrations detected in the stock solution were 101.0%, 100.0% and 112.0%, respectively, with RSD values of 2.6%, 4.0% and 0.0%, respectively. The recovery rates of various concentrations in finished product testing were 104.0%, 94.7% and 112.0%, respectively, with RSD values of 1.9%, 2.4% and 0.0%, respectively. As for the intermediate precision, the RSD were 4.5% and 3.7%, respectively. After treated with sample diluent, the sample was tested at room temperature for 6 hours and still exhibited relatively stable biological activity. [Conclusion] This detection method is accurate, stable, easy to operate and highly automated, and is suitable for detecting the potency of recombinant human coagulation factor Ⅶa for Injection.
2.Construction of PD-L1hitol-DC derived from bone marrow of DA rats and identification of its immunological function
Zhiqi YANG ; Peibo HOU ; Lang WU ; Jing LIU ; Yang DING ; Minghao LI
Organ Transplantation 2025;16(1):83-90
Objective To construct programmed cell death protein-ligand 1(PD-LI)hi tolerogenic dendritic cell (tol-DC) derived from bone marrow of DA rats and identify its immunological function. Methods DA rat bone marrow cells were extracted, combined with recombinant mouse granulocyte macrophage colony-stimulating factor and recombinant mouse interleukin (IL)-4, and cultured for 6 days in vitro to induce the differentiation of bone marrow cells into immature dendritic cells (imDC). Lipopolysaccharide was used to stimulate cell maturation and cultured for 2 days to collect mature dendritic cells (mDC). PD-L1 lentiviral vector virus stock solution or equivalent dose lentiviral stock solution was added, and PD-L1hitol-DC and Lv-imDC were collected after culture for 2 days. The morphology of PD-L1hitol-DC was observed by inverted phase contrast microscope and transmission electron microscope. Real-time fluorescence quantitative reverse transcription polymerase chain reaction, Western blotting and flow cytometry were used to detect the expression level of specific markers on cell surface. CD8+T cells derived from Lewis rat spleen were co-cultured with imDC, mDC, Lv-imDC and PD-L1hitol-DC, respectively. The levels of inflammatory factors in the supernatant of each group were detected by enzyme-linked immunosorbent assay. The apoptosis of T cells and the differentiation of regulatory T cells (Treg) in each group were analyzed by flow cytometry. Results The morphology of PD-L1hitol-DC modified by PD-L1 gene was consistent with tol-DC characteristics, and the expression levels of CD80, CD86 and major histocompatibility complex (MHC) on the surface were low. After mixed culture with CD8+ T cells, the levels of IL-10 and transforming growth factor (TGF) -β1 in the supernatant of PD-L1hitol-DC group were higher, the levels of tumor necrosis factor (TNF) -α and IL-17A were lower, and the apoptosis of T cells and Treg differentiation were increased. Conclusions Overexpression of PD-L1 through lentiviral vectors may successfully induce the construction of bone-marrow derived PD-L1hitol-DC in DA rats, promote the secretion of anti-inflammatory factors and T cell apoptosis, induce the differentiation of Treg, and inhibit the immune response of allogeneic CD8+T cells, which provides experimental basis for the next organ transplantation immune tolerance study.
3.Mechanism of inhibitory effect of total flavonoids from Taraxacum mongolicum on obesity in mice by regulating intestinal flora
Yixue GAO ; Lin GUO ; Linyan LANG ; Jing WU ; Haoyang WANG ; Jing YANG ; Mingsan MIAO ; Zhanzhan LI
China Pharmacy 2025;36(3):293-299
OBJECTIVE To investigate the mechanism of the inhibitory effect of total flavonoids from Taraxacum mongolicum on high-fat diet-induced obesity in mice through modulation of intestinal flora. METHODS Twenty-four C57BL/6J mice were randomly divided into blank group, model group and T. mongolicum total flavonoid group, with 8 mice in each group. Except for the blank group, the other 2 groups were given a high-fat diet, while T. mongolicum total flavonoid group was given T. mongolicum total flavonoid [400 mg/(kg·d)] intragastrically, once a day, for 8 consecutive weeks. During the experiment, the food intake of each group of mice was recorded. After the last medication, the body mass, fat weight, blood lipid level and pathological changes of liver and epididymal fat in mice were evaluated to observe the effect of T. mongolicum total flavonoid on the treatment of obesity in mice. The changes in abundance and structure of intestinal flora in mice were detected by amplicon sequencing; the effects of T. mongolicum total flavonoids on fat metabolism related genes were analyzed by qPCR. RESULTS Compared with model group, the body weight of mice in T. mongolicum total flavonoids group was decreased significantly (P<0.05); the levels of total lipid cholesterol, triglycerides, and LDL cholesterol were all decreased significantly (P<0.01), and the level of HDL cholesterol was increased significantly (P<0.01); the fat indexes of inguinal white adipose tissue and epididymal white wind_lz@hactcm.edu.cn adipose tissue were significantly reduced (P<0.05); significant improvement in hepatocellular steatosis and adipose cytopathy were significantly improved; mRNA expressions of COX7A1 and COX8B were significantly upregulated (P<0.05). The results of bacterial colony detection showed that compared with the model group, there was a rising trend in the diversity of the bacterial colony in T. mongolicum total flavonoids group, and the Sobs index characterization and β diversity were increased significantly (P<0.05). Relative abundances of Blautia, norank_f_Ruminococcaceae, Bilophila, Alistipes, classified_f_Ruminococcaceae, Parabacteroides, norank_f_Desulfovibrionaceae, Anaerotruncus were significantly up-regulated(P<0.05), while those of Faecalibaculum, Erysipelatoclostridium, GCA-900066575, Tuzzerella, Lactobacillus, norank_f_norank_o_RF39, achnospiraceae_FCS020_group were significantly down-regulated (P<0.05). CONCLUSIONS T. mongolicum total flavonoids can reduce body mass, fat weight and blood lipid levels, and repair the pathological damage to liver and epididymal fat in obese mice, which is related to improving intestinal flora disorders caused by high-fat diet.
4.Experience in Treating Acne Based on the Staged Approach of "Eruption in Warm Diseases"
Yisheng ZHANG ; Ningxin ZHANG ; Fengyan TIAN ; Yuanyao SHE ; Jing LANG ; Weili KONG ; Qingyun LIU
Journal of Traditional Chinese Medicine 2025;66(16):1723-1726
This paper summarizes clinical experience in treating acne based on the staged therapeutic principles of "eruption in warm diseases". It is considered that acne results from wind-heat retained in the lungs, invading the ying level and obstructing the blood collaterals, and is primarily a disorder involving both the wei and ying systems. In clinical practice, the treatment emphasizes the use of acrid-cool and sweet-cold methods. The core prescription is namely Yinqiaosan Qu Douchi Jia Xishengdi Danpi Daqingye Bei Xuanshen Fang (from Epidemic Warm Diseases [《温病条辨》]), and is adjusted according to the stage of disease. In the non-inflammatory stage, when the pathogen initially attacks the wei level, treatment focuses on acrid-cool herbs to release the exterior, with supplementary bitter-sweet ingredients such as Yejuhua (Chrysanthemum Indicum). In the inflammatory stage, with pronounced heat toxin in the qi level affecting the ying and blood, and local stagnation of qi and blood, the approach is to clear heat and resolve toxin, using blood-cooling and stasis-resolving herbs early to prevent progression. Herbs such as Pugongying (Taraxacum Mongolicum), Zihuadiding (Viola Yedoensis), Tiankuizi (Semiaquilegia Adoxoides), Chonglou (Paris Polyphylla), Machixian (Portulaca Oleracea), Zaojiaoci (Gleditsia Sinensis), Chuanshanjia (Manis Pentadactyla) may be added. In the post-inflammatory erythema stage, when yin of the ying level is depleted and internal deficiency-heat arises, sweet-cold herbs are recommended to nourish the stomach and generate fluids, with the possible addition of Yiwei Decoction (益胃汤).
6.The Spatial Differences and Dynamic Evolution of China's Healthcare Service Efficiency from 2012 to 2021
Sha-Sha SONG ; Lina SHAO ; Zhonghua SUO ; Jing WU ; Ying LANG
Chinese Health Economics 2024;43(9):70-74,96
Objective:To study the longitudinal trends and spatial clustering characteristics of healthcare service efficiency in China and in North,Northeast,East,Central,South,Southwest,and Northwest China.Methods:The Malmquist index model is used to measure China's healthcare service efficiency from 2012 to 2021,the Dagum Gini coefficient as well as the decomposition method are used to measure the magnitude and source of regional gaps in healthcare service efficiency,and the Kernel density estimation is used to study the longitudinal trend of change and spatial agglomeration characteristics of China's healthcare service efficiency.Results:China's overall healthcare service efficiency is growing,and the inter-regional gap is gradually narrowing,characterized by a concentration trend;the gap in the level of healthcare service efficiency between regions did not widen during the period under examination,but it was found that the gap within some regions was still significant.Conclusion:The national health service efficiency is growing slightly,and the regional gap is generally decreasing,but the Gini coefficient shows that the inter-regional contribution is still the main source of the gap.National health service efficiency is generally concentrated,but some regions are less efficient,with significant internal disparities.
7.Epidemiological characteristics and toxicant type of acute poisoning cases in China, 2016-2022
Yutao ZHANG ; Shaofeng JIANG ; Nan LANG ; Jing ZHOU ; Hongshun ZHANG
Chinese Journal of Epidemiology 2024;45(10):1376-1382
Objective:To analyze the acute poison epidemic and provide evidence for developing prevention and control strategies for acute poisoning.Methods:A retrospective analysis was conducted on acute poisoning cases collected from 2016 to 2022 in a health emergency information platform for acute poisoning accidents. The cases were grouped according to the distribution of poisoning occurrence time, geographic distribution, demographic distribution, types of toxicants, poisoning causes, and outcomes. Data were organized and analyzed using Excel 2016 and R 4.2.3.Results:A total of 95 754 acute poisoning cases were included in this study. The primary toxicants were pesticides, drugs, and industrial/household chemicals, accounting for 30.4%, 22.4%, and 20.4% of the total cases, respectively. Acute poisoning occurred throughout the year, with the highest frequency from June to August, accounting for 31.9%. The seasonal distribution varied among different types of toxicants. Except for plant poisoning, which showed a bimodal distribution, the other poisonings showed an unimodal distribution. There was a strong seasonality in fungal poisoning, which peaked in July. There was an obvious seasonality in animal poisoning, with a peak in August. The proportion of biological poisonings in the southwest region was higher than in other regions, including plants, animals, and fungi. There were more females than males, and their education level was mainly junior high school and below (35.2%). The main occupation was farmers (34.2%), and the main causes of poisoning were accidents and suicides. The case fatality rate of all poisoning cases was 1.24%. Pesticide poisoning was the most common type, and chlorfenapyr (11.68%), Diquat (7.23%), and paraquat (7.05%) ranked as the top three toxicants.Conclusions:The occurrence of acute poisoning has an obvious seasonal trend, and the toxicant spectrum of different regions and populations is different. A comprehensive poisoning surveillance system can provide a better understanding of the occurrence of poisonings, and facilitate the formulation of more scientifically precise poisoning prevention and control strategies.
8.Spermidine alleviates pressure overload-induced heart failure in mice via improving cardiac mitochondrial energy metabolism
Xiaoliang ZHANG ; Xiaoling ZHAO ; Jing GENG ; Lang HU ; Yan LI
Chinese Journal of Pathophysiology 2024;40(2):193-203
AIM:To investigate the effect of spermidine(SPD)on pressure overload-induced cardiac hyper-trophy and heart failure model in mice and its underlying mechanisms.METHODS:(1)Eight-week-old male C57BL/6J mice were randomly divided into 4 groups:sham group,sham+SPD group,transverse aortic constriction(TAC)group,and TAC+SPD group.After TAC,the mice in sham+SPD group and TAC+SPD group were fed with 3 mmol/L SPD via drinking water,and the mice in other groups were fed with normal water.Western blot was used to detect the protein ex-pression levels of silent information regulator 6(SIRT6),peroxisome proliferator-activated receptor γ coactivator-1(PGC-1)and mitofusin 2(MFN2).Adult mouse cardiomyocytes were isolated to detect cell length and width.Wheat germ agglu-tinin staining was used to detect the cardiac cell size.Masson staining was used to detect the extent of fibrosis.Echocar-diography was used to detect cardiac function and myocardial hypertrophy.Transmission electron microscopy was used to analyze mitochondrial morphology.Oxygraph-2k high-resolution respirometer was used to detect cardiac mitochondrial oxy-gen consumption.(2)In vitro,primary rat ventricular cardiomyocytes were cultured and treated with angiotensin II(Ang II;1 μmol/L)to construct a hypertrophy model of cardiomyocytes.These cardiomyocytes were divided into control(Con)group,Con+SPD(1 mmol/L)group,Ang II group,Ang II+SPD group and Ang II+SPD+SIRT6 siRNA(siSIRT6)group.Confocal microscopy was used to detect cardiomyocytes area and mitochondrial.RESULTS:(1)Compared with sham group,cardiac function of the mice in TAC group was significantly decreased(P<0.05),the degree of myocardial hyper-trophy was significantly increased(P<0.05),and the expression levels of SIRT6,PGC-1 and MFN2 in the myocardial tis-sue were significantly decreased(P<0.05).Compared with TAC group,the expression levels of SIRT6,PGC-1 and MFN2 in mouse myocardial tissues of TAC+SPD group were significantly increased(P<0.05),pathological myocardial hy-pertrophy was reduced(P<0.05),the numbers of mitochondria and mitochondrial cristae were increased(P<0.05),mito-chondrial function was restored(P<0.05),myocardial fibrosis was alleviated(P<0.05),and cardiac function was im-proved(P<0.05).(2)In vitro,compared with Con group,the expression levels of SIRT6,PGC-1 and MFN2 in cardio-myocytes of Ang II group were decreased(P<0.05),and the degree of cardiomyocyte hypertrophy was significantly in-creased(P<0.05).Treatment with SPD increased the expression levels of SIRT6,PGC-1 and MFN2 in cardiomyocytes of Ang II group(P<0.05),reversed myocardial hypertrophy and improved mitochondrial dynamics(P<0.05).Compared with Ang II group,the expression levels of SIRT6,PGC-1 and MFN2 in Ang II+SPD+siSIRT6 group showed no significant changes,and the degree of cardiomyocyte hypertrophy and mitochondrial dynamics also had no statistically significant changes.CONCLUSION:Spermidine promotes the expression of SIRT6,PGC-1 and MFN2,thus improving mitochon-drial function,reducing myocardial hypertrophy and alleviating heart failure in mice with pressure overload.
9.Correlation analysis of miR-4500,NGF expressions with preoperative magnetic resonance signs in breast cancer
Fan YANG ; Jing CHEN ; Bin ZHENG ; Jiao-Jiao LI ; Li ZHANG ; Gui-Feng BIAN ; Ya-Jing BIAN ; Xiao-Yan LANG
Journal of Regional Anatomy and Operative Surgery 2024;33(1):47-50
Objective To analyze the correlations of the expressions of miR-4500 and nerve growth factor(NGF)in breast cancer patients with preoperative magnetic resonance(MR)signs.Methods A total of 105 patients with breast cancer admitted to our hospital were selected,the mRNA expression levels of miR-4500 and NGF in breast cancer tissues and adjacent tissues of patients were detected by qRT-PCR,and the positive expression rates of NGF in breast cancer tissues and adjacent tissues were determined by immunohistochemistry method.The correlations of the expressions of miR-4500 and NGF in breast cancer tissues with clinicopathological features and MR signs of patients were analyzed.The targeting relationship between miR-4500 and NGF was predicted by the bioinformatics website,and the correla-tion between the expressions of the two was analyzed.Results Compared with the adjacent tissues,the expression level of miR-4500 in breast cancer tissue decreased(P<0.05),while the level of NGF mRNA and the positive expression rate of NGF increased(P<0.05).There was no significant difference in age,pathological type,tumor classification,parenchymal background,apparent diffusion coefficient or enhancement curve among different expressions of miR-4500 and NGF(P>0.05).The histological grade,lymph node metastasis,tumor diameter,tumor morphology,ring-like enhancement,and peritu-moral brain edema were related to the expressions of miR-4500 and NGF(P<0.05).The bioinformatics website prediction showed that miR-4500 and NGF had binding sites,and the expressions of miR-4500 and NGF mRNA in breast cancer tissues were negatively correlated(r=-0.576,P<0.05).Conclusion The expression of miR-4500 in breast cancer tissue is low,and the expression of NGF is high,which are correlated with the preoperative MR signs in patients,providing a molecu-lar basis for MR signs.
10.Herpesvirus encephalomyelitis with the main symptom of myelitis:A case report
Xingjun LI ; Yue LANG ; Jing BAI
Journal of Apoplexy and Nervous Diseases 2024;41(10):941-944
Herpesvirus encephalomyelitis is an infectious disease of the central nervous system,and there are rare cases of herpesvirus encephalitis comorbid with myelitis.This article reports a case of herpesvirus encephalomyelitis with the main symptom of myelitis,as well as other symptoms of headache,immobility of both lower limbs,and dysurea.Related clinical data and diagnosis and treatment processes are analyzed,and a literature review was performed to improve the understanding of this disease among clinicians.

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