1.Extraction and identification of primary rat brain microvascular endothelial cells by improved tissue block culture method
Fan Zhang ; Bolin Li ; Ming Chi ; Haiqin Liu ; Yuanyu Tang
Acta Universitatis Medicinalis Anhui 2025;60(1):10-14
Objective :
To investigate the brain microvascular tissue block culture method for extracting primary rat brain microvascular endothelial cells and identify its effect.
Methods :
Brain tissue from 4-week-old Sprague Dawley rats was screened, pre-digested and solidified to obtain brain microvascular segments. These segments were subsequently placed in a CO2incubator for primary culture. The target cells were identified by cell morphology and immunocytochemical staining for factor Ⅷ-related antigen.
Results :
After a 48-hour culture periodin vitro, the short spindle cells crawled out from around the brain microvascular segments. After 72 hours, island-like cell culsters formed. After 96 hours the clusters fused and the cells formed a typical monolayer, cobble stone-like, and mosaic arrangement. Factor Ⅷ-related antigen immunocytochemical staining showed that the cytoplasm of the cells appeared brown-red, indicating positive expression; DAB stained the nucleus, showing blue-dark.
Conclusion
The brain microvascular tissue block culture method can isolate and culture primary rat brain microvascular endothelial cells.
2.Rehmanniae Radix Iridoid Glycosides Protect Kidneys of Diabetic Mice by Regulating TGF-β1/Smads Signaling Pathway
Hongwei ZHANG ; Ming LIU ; Huisen WANG ; Wenjing GE ; Xuexia ZHANG ; Qian ZHOU ; Huani LI ; Suqin TANG ; Gengsheng LI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(2):56-66
ObjectiveTo investigate the protective effect of Rehmanniae Radix iridoid glycosides (RIG) on the kidney tissue of streptozotocin (STZ)-induced diabetic mice and explore the underlying mechanism. MethodsTwelve of 72 male C57BL/6J mice were randomly selected as the normal group, and the remaining 60 mice were fed with a high-fat diet for six weeks combined with injection of 60 mg·kg-1 STZ for 4 days to model type 2 diabetes mellitus. The successfully modeled mice were randomized into model, metformin (250 mg·kg-1), catalpol (100 mg·kg-1), low-dose RIG (RIG-L, 200 mg·kg-1) and high-dose RIG (RIG-H, 400 mg·kg-1) groups (n=11). Mice in each group were administrated with corresponding drugs, while those in the normal group and model group were administrated with the same dose of distilled water by gavage once a day. After 8 weeks of intervention, an oral glucose tolerance test (OGTT) was performed, and the area under the curve (AUC) was calculated. After mice were sacrificed, both kidneys were collected. The body weight, kidney weight, and fasting blood glucose (FBG) were measured. Biochemical assays were performed to measure the serum levels of triglycerides (TG), total cholesterol (TC), serum creatinine (SCr), and blood urea nitrogen (BUN). Enzyme-linked immunosorbent assay (ELISA) was employed to determine the serum level of fasting insulin (FINS), and the insulin sensitivity index (ISI) and homeostatic model assessment for insulin resistance (HOMA-IR) were calculated. The pathological changes in kidneys of mice were observed by hematoxylin-eosin staining and Masson staining. The immunohistochemical method (IHC) was employed to assess the expression of interleukin-1 (IL-1), interleukin-6 (IL-6), tumor necrosis factor-α(TNF-α), transforming growth factor-β1 (TGF-β1), and collagen-3 (ColⅢ) in the kidney tissue. The protein levels of TGF-β1, cell signal transduction molecule 3 (Smad3), matrix metalloproteinase-9 (MMP-9), and ColⅢ in kidneys of mice were determined by Western blot. ResultsCompared with the normal group, the model group showcased decreased body weight and ISI (P<0.01), increased kidney weight, FBG, AUC, FINS, HOMA-IR, TC, TG, SCr, and BUN (P<0.01), glomerular hypertrophy, capsular space narrowing, and collagen deposition in the kidney, up-regulated protein levels of IL-1, IL-6, TNF-α, TGF-β1, ColⅢ, and Smad3 (P<0.01), and down-regulated protein level of MMP-9 (P<0.01) in the kidney tissue. Compared with the model group, the treatment groups had no significant difference in the body weight and decreased kidney weight (P<0.05, P<0.01). The FBG level declined in the RIG-H group after treatment for 4-8 weeks and in the metformin, catalpol, and RIG-L groups after treatment for 6-8 weeks (P<0.01). The AUC in the RIG-L, RIG-H, and metformin groups decreased (P<0.05, P<0.01). The levels of TC, SCr, and BUN in the serum of mice in each treatment group became lowered (P<0.05, P<0.01). The level of TG declined in the RIG-L, RIG-H, and metformin groups (P<0.05, P<0.01). The serum level of FINS declined in the catalpol, RIG-L, and metformin groups (P<0.01). Compared with the model group, the treatment groups showed decreased HOMA-IR (P<0.01), increased ISI (P<0.01), alleviated pathological changes in the kidney tissue, and down-regulated expression of IL-1 and TGF-β1. In addition, the protein levels of IL-6, TNF-α, and ColⅢ in the RIG-H and metformin groups and IL-6 and TNF-α in the RIG-L group were down-regulated (P<0.05, P<0.01), and the protein levels of IL-6, TNF-α, and ColⅢ in the catalpol group and ColⅢ in the RIG-L group showed a decreasing trend without statistical difference. The protein levels of TGF-β1, Smad3, and ColⅢ in the RIG-H and metformin groups were down-regulated (P<0.01). Compared with that in the model group, the protein level of MMP-9 was up-regulated in each treatment group (P<0.01). ConclusionRIG can improve the renal structure and function of diabetic mice by regulating the TGF-β1/Smads signaling pathway.
3.Preliminary application of sacral neuromodulation in patients with benign prostatic hyperplasia complicated with underactive bladder after transurethral resection of the prostate
Ning LIU ; Yan ZHANG ; Tao LI ; Qiang HU ; Kai LU ; Lei ZHANG ; Jianping WU ; Shuqiu CHEN ; Bin XU ; Ming CHEN
Journal of Modern Urology 2025;30(1):39-42
[Objective] To evaluate the efficacy and safety of sacral neuromodulation (SNM) in the treatment of patients with benign prostatic hyperplasia (BPH) complicated with underactive bladder (UAB) who respond poorly to transurethral resection of the prostate (TURP). [Methods] A retrospective analysis was performed on 10 patients with BPH and UAB treated with TURP by the same surgeon in Zhongda Hospital Southeast University during Jan.2018 and Jan.2023.The residual urine volume was not significantly relieved after operation, and the maximum urine flow rate and urine volume per discharge were not significantly improved.All patients underwent phase I SNM, and urinary diaries were recorded before and after surgery to observe the average daily frequency of urination, volume per urination, maximum urine flow rate, and residual urine volume. [Results] The operation time was (97.6±11.2) min.During the postoperative test of 2-4 weeks, if the residual urine volume reduction by more than 50% was deemed as effective, SNM was effective in 6 patients (60.0%). Compared with preoperative results, the daily frequency of urination [(20.2±3.8) times vs. (13.2±3.2) times], volume per urination [(119.2±56.7) mL vs. (246.5±59.2) mL], maximum urine flow rate [(8.7±1.5) mL/s vs. (16.5±2.6) mL/s], and residual urine volume [(222.5±55.0) mL vs. (80.8±16.0) mL] were significantly improved, with statistical significance (P<0.05). There were no complications such as bleeding, infection, fever or pain.The 6 patients who had effective outcomes successfully completed phase II surgery, and the fistula was removed.During the follow-up of 1 year, the curative effect was stable, and there were no complications such as electrode displacement, incision infection, or pain in the irritation sites.The residual urine volume of the other 4 unsuccessful patients did not improve significantly, and the electrodes were removed and the vesicostomy tube was retained. [Conclusion] SNM is safe and effective in the treatment of BPH with UAB patients with poor curative effects after TURP.
4.Pharmacokinetic study of 3 blood-absorbed components of Xiangshao sanjie oral liquid in rats with hyperplasia of mammary gland
Yu ZHANG ; Jiaming LI ; Dan PENG ; Ruoqiu FU ; Yue MING ; Zhengbi LIU ; Jingjing WANG ; Shiqi CHENG ; Hongjun XIE ; Yao LIU
China Pharmacy 2025;36(6):680-685
OBJECTIVE To explore the pharmacokinetic characteristics of 3 blood-absorbed components of Xiangshao sanjie oral liquid in rats with hyperplasia of mammary gland (HMG). METHODS Female SD rats were divided into control group and HMG group according to body weight, with 6 rats in each group. The HMG group was given estrogen+progesterone to construct HMG model. After modeling, two groups were given 1.485 g/kg of Xiangshao sanjie oral liquid (calculated by crude drug) intragastrically, once a day, for 7 consecutive days. Blood samples were collected before the first administration (0 h), and at 5, 15, 30 minutes and 1, 2, 4, 8, 12, 24 hours after the last administration, respectively. Using chlorzoxazone as the internal standard, the plasma concentrations of ferulic acid, paeoniflorin and rosmarinic acid in rats were detected by UPLC-Q/TOF-MS. The pharmacokinetic parameters [area under the drug time curve (AUC0-24 h, AUC0-∞), mean residence time (MRT0-∞), half-life (t1/2), peak time (tmax), peak concentration (cmax)] were calculated by the non-atrioventricular model using Phoenix WinNonlin 8.1 software. RESULTS Compared with the control group, the AUC0-24 h, AUC0-∞ and cmax of ferulic acid in the HMG group were significantly increased (P<0.05); the AUC0-24 h, AUC0-∞ , MRT0-∞ , t1/2 and cmax of paeoniflorin increased, but there was no significant difference between 2 groups (P>0.05); the AUC0-24 h and MRT0-∞ of rosmarinic acid were significantly increased or prolonged (P<0.05). C ONCLUSIONS In HMG model rats, the exposure of ferulic acid, paeoniflorin and rosmarinic acid in Xiangshao sanjie oral liquid all increase, and the retention time of rosmarinic acid is significantly prolonged.
5.Terms Related to The Study of Biomacromolecular Condensates
Ke RUAN ; Xiao-Feng FANG ; Dan LI ; Pi-Long LI ; Yi LIN ; Zheng WANG ; Yun-Yu SHI ; Ming-Jie ZHANG ; Hong ZHANG ; Cong LIU
Progress in Biochemistry and Biophysics 2025;52(4):1027-1035
Biomolecular condensates are formed through phase separation of biomacromolecules such as proteins and RNAs. These condensates exhibit liquid-like properties that can futher transition into more stable material states. They form complex internal structures via multivalent weak interactions, enabling precise spatiotemporal regulations. However, the use of inconsistent and non-standardized terminology has become increasingly problematic, hindering academic exchange and the dissemination of scientific knowledge. Therefore, it is necessary to discuss the terminology related to biomolecular condensates in order to clarify concepts, promote interdisciplinary cooperation, enhance research efficiency, and support the healthy development of this field.
6.Analysis of Clinical Consistency of Animal Models of Attention Deficit Hyperactivity Disorder Based on Characteristics of Clinical Diseases and Syndromes in Traditional Chinese and Western Medicine
Xinyue XIE ; Xiaomian LIU ; Ming LI ; Mengfei WANG ; Rongyi ZHOU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(13):270-278
ObjectiveBased on a new method for animal model evaluation, this study aims to analyze the characteristics of diseases and syndromes of existing animal models of attention deficit hyperactivity disorder (ADHD) from both traditional Chinese medicine (TCM) and western medical perspectives and propose suggestions for improvement. MethodsA systematic search of the China National Knowledge Infrastructure (CNKI) and PubMed was conducted for literature on ADHD animal models. According to TCM and western medical diagnostic criteria, core and accompanying symptoms of the models were assigned with scores to comprehensively evaluate the clinical consistency. ResultsThe selection of experimental animals for ADHD models primarily involved rodents, with modeling methods including genetic, chemical induction, and environmental induction. The average consistency of clinical diseases and syndromes with TCM and western medicine was 45.19% and 49.42%, respectively. The spontaneously hypertensive (SHR) rats and nicotine/smoking models had the highest consistency with TCM, while the social isolation models had the highest consistency with western medicine. Most models were guided by western medicine theories, which can meet the surface validity and structural validity requirements of western medicine but lacked precise differentiation of TCM syndromes. ConclusionExisting ADHD animal models primarily focus on a single genotype or environmental factor, lacking comprehensive consideration of multigenic interactions and environmental factors. Moreover, the selection of model evaluation indicators is relatively singular, primarily focusing on "disease" indicators, while TCM "syndrome" indicators have not been fully considered. It is recommended to introduce a "formula-to-syndrome" approach in the preparation of TCM models for ADHD and establish and improve an evaluation system of animal models combining diseases and syndromes, so as to provide a solid foundation for future experimental research.
7.Analysis of Clinical Consistency of Animal Models of Attention Deficit Hyperactivity Disorder Based on Characteristics of Clinical Diseases and Syndromes in Traditional Chinese and Western Medicine
Xinyue XIE ; Xiaomian LIU ; Ming LI ; Mengfei WANG ; Rongyi ZHOU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(13):270-278
ObjectiveBased on a new method for animal model evaluation, this study aims to analyze the characteristics of diseases and syndromes of existing animal models of attention deficit hyperactivity disorder (ADHD) from both traditional Chinese medicine (TCM) and western medical perspectives and propose suggestions for improvement. MethodsA systematic search of the China National Knowledge Infrastructure (CNKI) and PubMed was conducted for literature on ADHD animal models. According to TCM and western medical diagnostic criteria, core and accompanying symptoms of the models were assigned with scores to comprehensively evaluate the clinical consistency. ResultsThe selection of experimental animals for ADHD models primarily involved rodents, with modeling methods including genetic, chemical induction, and environmental induction. The average consistency of clinical diseases and syndromes with TCM and western medicine was 45.19% and 49.42%, respectively. The spontaneously hypertensive (SHR) rats and nicotine/smoking models had the highest consistency with TCM, while the social isolation models had the highest consistency with western medicine. Most models were guided by western medicine theories, which can meet the surface validity and structural validity requirements of western medicine but lacked precise differentiation of TCM syndromes. ConclusionExisting ADHD animal models primarily focus on a single genotype or environmental factor, lacking comprehensive consideration of multigenic interactions and environmental factors. Moreover, the selection of model evaluation indicators is relatively singular, primarily focusing on "disease" indicators, while TCM "syndrome" indicators have not been fully considered. It is recommended to introduce a "formula-to-syndrome" approach in the preparation of TCM models for ADHD and establish and improve an evaluation system of animal models combining diseases and syndromes, so as to provide a solid foundation for future experimental research.
8.Simulation analysis of the protective performance of barium sulfate mortar against positron nuclide γ-rays
Zhiqiang XU ; Huaixin NI ; Jiwu GENG ; Lichun LI ; Zaoqin ZHANG ; Shibiao SU ; Meixia WANG ; Ming LIU
Chinese Journal of Radiological Health 2025;34(2):209-213
Objective To obtain the protective performance parameters of barium sulfate mortar against positron nuclide γ-rays, provide reference data for precise shielding calculations, and guide the design, evaluation, and construction of radiation shielding. Methods The FLUKA program was used to build a model for simulating the dose equivalent rate variation around points of interest under the irradiation of the most commonly used positron nuclide 18F with changes in the thicknesses of lead and barium sulfate mortar. The transmission curves of lead and barium sulfate mortar were fitted, and the half-value layer (HVL) and lead equivalence of barium sulfate mortar were calculated based on the fitted curves. Results The ambient dose equivalent rate coefficient of positron nuclide 18F was 1.339 4×10−1 μSv·m2/MBq·h and the HVL for lead was 4.037 mm, with deviations of 0.043% and 1.53% compared to the values provided in the AAPM Report No. 108, respectively. The HVLs for γ-rays produced by 18F, using barium sulfate mortar with apparent densities of 4.20, 4.00, and 3.90 g/cm3 mixed with 35.2-grade cement in a 4∶1 mass ratio, were 2.914, 2.969, and 3.079 cm, respectively. The lead equivalences were
9.Feasibility of Multiparameter MRI-Guided Percutaneous Biopsy for Central Lung Lesions With Atelectasis
Peipei LI ; Chengli LI ; Yujun XU ; Xiangmeng HE ; Roberto Blanco SEQUEIROS ; Ming LIU
Korean Journal of Radiology 2025;26(5):498-507
Objective:
To prospectively evaluate the feasibility, accuracy, and safety of multiparameter MRI-guided percutaneous biopsy using a 1T open MRI scanner for evaluating suspicious centrally located lung lesions with associated post-obstructive atelectasis.
Materials and Methods:
In this single-center study, MRI-guided percutaneous coaxial cutting biopsy was performed for 107 suspicious central lung lesions with associated post-obstructive atelectasis in 107 patients between July 2015 and December 2020. A fast T2-weighted imaging (T2WI)-turbo spin echo (TSE) sequence and an enhanced fast T1-weighted imaging (T1WI)-TSE sequence were used to identify, localize, and biopsy lung lesions, and diffusion-weighted imaging (DWI) was used as a supplementary sequence for identifying the lesion location. The final diagnosis was confirmed by surgical histopathology or clinical follow-up for a minimum of 24 months. The sensitivity, specificity, and accuracy for diagnosing lung malignancies were calculated, and the complications were recorded for each case.
Results:
Using multiparameter MRI, central lung lesions could be clearly distinguished from post-obstructive atelectasis in 96 patients (89.7%). The sensitivity, specificity, and accuracy of MRI-guided percutaneous biopsy for diagnosing lung malignancy was 97.0% (98/101), 100% (6/6), and 97.2% (104/107), respectively. Self-limited hemoptysis occurred in three patients. Pneumothorax occurred in five patients, of which none required pleural drainage. No serious procedure-related complications were observed.
Conclusion
As a technology that does not involve ionizing radiation, multiparameter MRI-guided percutaneous coaxial cutting biopsy is a safe and accurate diagnostic technique for evaluating centrally located lung lesions associated with post-obstructive atelectasis.
10.Causal association of cathepsins with female infertility: a bidirectional Mendelian randomization analysis
Lidan LIU ; Ming LIAO ; Bo LIU ; Qianyi HUANG ; Huimei WU ; Mujun LI
Obstetrics & Gynecology Science 2025;68(3):237-243
Objective:
This study aimed to systematically evaluate potential causal relationships between nine cathepsins and female infertility using Mendelian randomization (MR) methods.
Methods:
A bidirectional MR analysis was conducted utilizing single nucleotide polymorphisms as instrumental variables to investigate the potential causal effects between nine cathepsins and female infertility. Genetic data on female infertility were sourced from the FinnGen study, and cathepsin-related data were obtained from genome-wide association studies datasets of European ancestry.
Results:
Elevated levels of cathepsin E were significantly and inversely associated with the risk of female infertility, suggesting a potential protective role. This finding was further supported by multivariable MR analysis. However, no significant associations were observed between the other eight cathepsins and female infertility.
Conclusion
This study represents the first systematic MR analysis to identify a potential protective effect of cathepsin E on female infertility.


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