1.Molecular Crosstalk Mechanisms of Shoutai Wan and Juyuan Jian on Maternal-fetal Interface Subcellular Clusters in CBA/J×DBA/2 Recurrent Pregnancy Loss Model
Jingxin GAO ; Qiuping CHEN ; Xiaoyan ZHENG ; Pengfei ZENG ; Rui ZHOU ; Yancai TANG ; Qian ZENG ; Wenli GUO ; Jinzhu HUANG ; Weijun DING ; Linwen DENG ; Hang ZHOU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(2):70-87
ObjectiveTo systematically compare the differential regulation of the maternal-fetal interface cell lineages and communication networks in the CBA/J×DBA/2 mouse model of recurrent pregnancy loss (RPL) by the two classic therapeutic methods-tonifying the kidney to stabilize the fetus and invigorating the spleen to stabilize the fetus (Shoutai Wan, Juyuan Jian)-of traditional Chinese medicine (TCM) at the single-cell resolution and clarify their modern scientific connotations. MethodsFemale non-pregnant CBA/J mice were caged with male BALB/c (blank group) and DBA/2 (modeling group) mice separately. Pregnant mice in the modeling group were randomly grouped as follows: high/low-dose Shoutai Wan, high/low-dose Juyuan Jian, model (RPL), and positive control (dydrogesterone), with 10 mice in each group. Starting from the day after the detection of the vaginal plug, mice were administrated with drugs or an equal volume of normal saline by gavage for 10 consecutive days. After the intervention, the following indicators were measured. ① Macroscopic evaluation: general conditions, uterine wet weight, embryo loss rate, four coagulation parameters [prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (FIB), and thrombin time (TT)], and peripheral blood estradiol (E2) and progesterone (Pg) levels. The decidua with embryos was stained with hematoxylin-eosin (HE) and evaluated by transmission electron microscopy (TEM). The expression of B-cell lymphoma-2 (Bcl-2), vascular endothelial growth factor (VEGF), angiotensin Ⅱ (AngⅡ), matrix metalloproteinase-2 (MMP-2), interleukin-6 (IL-6), leukemia inhibitory factor (LIF), CXC chemokine ligand 12 (CXCL12), and microtubule-associated protein 1 light chain 3 homolog (LC3)Ⅰ/Ⅱ was quantified by Western blot. ② Mechanism analysis at the single-cell level: The decidua with embryos from the blank, model, high-dose Shoutai Wan, and high-dose Juyuan Jian groups (6 mice per group, with 3 single-cell samples per group, totaling 24 mice) were analyzed by the BD Rhapsody™ platform, and the whole-cell atlas was drawn by uniform manifold approximation and projection (UMAP) dimensionality reduction clustering combined with the single-cell mouse cell atlas (scMCA). The differentially expressed genes (DEGs) and cell interaction networks were analyzed via Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and CellChat, and the protein-protein interaction (PPI) map of subtype cells was constructed. The CytoTRACE pseudo-temporal analysis was performed to explore the developmental trajectories of core immune cells (natural killer cells, NK cells) from maternal and fetal sources. Results① Pathological and Western blot results indicated that compared with the blank group, the RPL group showed an increase in the embryo loss rate (P<0.01), down-regulated expression of Bcl-2, LIF, MMP-2, and Vegf in the decidua with embryos (P<0.05), up-regulated protein levels of CXCL-12, AngⅡ, and IL-6 (P<0.05), blocked angiogenesis, apoptosis-inflammation imbalance, and coagulation dysfunction. Both prescriptions dose-dependently reduced the abortion rate and restored the angiogenesis-inflammation balance, and Shoutai pill showed superior performance in restoring the E2 level to the Pg level (P<0.05). ② Single-cell transcriptome analysis indicated that compared with the blank group, the RPL group showed differences in multiple key cell populations such as decidual cells, trophoblast cells, endothelial cells, erythroblasts, NK cells, and macrophages at the maternal-fetal interface. Immunity and angiogenesis were the key links in RPL. Compared with the RPL group, high-dose Shoutai Wan reversed the changes of NK cells in the embryonic layer (upregulating the mRNA levels of 17 genes and downregulating the mRNA levels of 29 genes) and macrophages (upregulating the mRNA levels of 117 genes and downregulating the mRNA levels of 53 genes) through the regulation of gene expression. High-dose Shoutai pill regulated the immune cells to affect unfolded proteins, cell adhesion, and programmed cell death, thereby promoting decidualization and angiogenesis and modulating embryo-membrane development. High-dose Juyuan Jian regulated the key subgroups of NK cells (up-regulating the mRNA levels of 9 genes and down-regulating the mRNA levels of 17 genes) and macrophages (up-regulating the mRNA levels of 110 genes and down-regulating the mRNA levels of 81 genes), which affected decidual inflammation and apoptosis and intervened in glycolysis. ③ The pseudo-temporal analysis and communication network indicated that the communication frequency of the RPL group decreased. High-dose Shoutai Wan restored maternal-fetal tolerance through pathways such as NKG2D, CDH5, GDF, and FASLG. High-dose Juyuan Jian enhanced the IL-6/LIFR/JAK/signal transducer and activator of transcription 3 (STAT3) and desmosome/SEMA6/tumor necrosis factor-like weak inducer of apoptosis (TWEAK) signaling to improve endometrial receptivity. The RPL group showed an increased proportion of toxic dNK7, a decreased proportion of reparative dNK4, and blocked embryo fNK1. High-dose Shoutai Wan down-regulated dNK7 and up-regulated dNK4. High-dose Juyuan Jian inhibited the terminal differentiation of dNK7 and up-regulated LILRB1, thus restoring the balance of cytotoxicity and repair. ConclusionBoth the kidney-tonifying and spleen-invigorating methods are effective in treating RPL. NK and macrophages are the key immune cells in the interaction between the embryo and the membrane. The kidney-tonifying method (Shoutai Wan) has an advantage in regulating the phenotypes of unfolded protein, cell adhesion, and programmed cell death, and shows expression characteristics closer to the physiological state in the regulation of NKG2D and CDH5 signals. The spleen-invigorating method (Juyuan Jian) has an advantage in regulating epithelial-mesenchymal transition (EMT), angiogenesis, and glycolysis and shows higher communication intensity in the IL-6 and LIFR pathways.
2.Analysis of influential factors for rivaroxaban-induced bleeding events in patients with non-valvular atrial fibril‐ lation complicated with coronary heart disease
Mingyu CHEN ; Min CHEN ; Jinzhu DENG ; Qiang DAI ; Hongjin GAO
China Pharmacy 2024;35(18):2272-2277
OBJECTIVE To analyze the influential factors for rivaroxaban-induced bleeding events in patients with non- valvular atrial fibrillation (NVAF) and coronary heart disease. METHODS A total of 64 hospitalized patients with NVAF complicated with coronary heart disease who were treated with rivaroxaban and admitted to the Fuzhou University Affiliated Provincial Hospital from November 2021 to May 2023 were included in this retrospective study. The demographic data, laboratory test indexes and other general clinical data, and steady-state trough concentration of rivaroxaban were collected, and the dose- adjusted trough concentration was calculated. The occurrence of bleeding events within 6 months after discharge was recorded. The univariate analysis and binary Logistic regression analysis were adopted to determine the independent risk factors of rivaroxaban- related bleeding events. The binary Logistic regression equation was constructed to predict the probability of bleeding events. The area under the receiver operator characteristic (ROC) curve (AUC) was used to analyze the predictive value of the regression equation. RESULTS Among 64 patients, 19 patients had 24 case-times bleeding events, most of which were mild bleeding (19 case-times, 79.2%), and mainly gastrointestinal bleeding (17 case-times, 70.8%). After symptomatic treatment and adjustment of the anticoagulant regimen, most of them were improved or cured. In the univariate analysis, the proportion of patients with a history of anemia, platelet count, urea nitrogen content, steady-state trough concentration of rivaroxaban, dose-adjusted trough concentration and coagulation indexes [international normalized ratio, prothrombin time (PT), activated partial thromboplastin time] in bleeding group were significantly more or higher than those in non-bleeding group, while the albumin level was significantly lower than that in non-bleeding group (P<0.05). In binary Logistics regression analysis, high PT level (odds ratio=1.473, 95% confidence interval=1.103-1.967, P= 0.009) and high rivaroxaban dose-adjusted trough concentration (odds ratio=1.174, 95% confidence interval=1.018-1.355, P= 0.027) were independent risk factors for rivaroxaban-related bleeding events. The binary Logistic regression equation of bleeding event prediction probability (P) was LogitP=-6.975+0.387×PT level+0.161×dose-adjusted trough concentration, and the AUC of the ROC curve was 0.825 (95% confidence interval was 0.708-0.909, P<0.001). CONCLUSIONS The risk factors of rivaroxaban-related bleeding events in patients with NVAF and coronary heart disease include previous anemia history, high platelet count, high urea nitrogen content, high rivaroxaban steady-state trough concentration, high dose-adjusted trough concentration, high coagulation indexes and low albumin level. High PT level and high dose-adjusted trough concentration are independent risk factors that can be used to predict the risk of rivaroxaban-induced bleeding events. The regression equation has good predictive value.
3.Determination of antioxidant activity of Polygoni Multiflori Radix Praeparata by HPLC-DPPH
Jinzhu QIAN ; Ming LIANG ; Yi YU ; Yalei DENG ; Shijia LIU ; Huiming PANG ; Jin QI
Journal of China Pharmaceutical University 2022;53(2):185-191
An on-line HPLC-DPPH system was developed to determine the antioxidant activity of 16 batches of Polygoni Multiflori Radix Praeparata. By analyzing the chromatographic and biological activity fingerprints of 16 batches of Polygoni Multiflori Radix Praeparata, the dose-effect relationship was established and the total antioxidant activity was quantified by activity addition.The results suggested that the online HPLC-DPPH method can evaluate the antioxidant activity of different bathches of Polygoni Multiflori Radix Praeparata, with different processing methods, aiming to provide datasupport and scientific basis forquality evaluation of Polygoni Multiflori Radix Praeparata.
4.Effect of External Method of Tonifying Kidney and Promoting Blood Circulation to Endometrial Morphology of Rats with Kidney Deficiency and Blood Stasis Model during Implantation Window
Jing LAN ; Jinzhu HUANG ; Linwen DENG ; Tianping WANG ; Yi WANG ; Qian ZENG
World Science and Technology-Modernization of Traditional Chinese Medicine 2015;(8):1700-1706
This study aimed to analyze the effect of the external method of tonifying kidney and promoting blood circulation on endometrial morphology of rats with kidney deficiency and blood stasis model. A total of 50 normal healthy unmated female SD rats of proestrus were selected with vaginal smear. Rats were randomly divided into the blank group, model group, high dose group and low dose group with the prescription of tonifying kidney and promoting blood circulation and theWu-Zi (WZ) group. Except the blank group, intragastric administration of hydroxyurea was given to other group to establish the kidney deficiency model. Meanwhile, clyster of distilled water was given to the blank group and the model group. And clyster of high dose and low dose prescription of tonifying kidney and promoting blood circulation was given to the high dose and low dose group, respectively. The intragastric administrations ofWu-Zi Yan-Zong(WZYZ) pills were given to theWu-Zi (WZ) group. On the 4th day of pregnancy, 10% of the macromolecule dextran was quickly injected to the caudal vein to induce blood stasis model 1 hour after the last medication administration. The uterus tissue section was observed by HE dyeing. The results showed that the thickness of covering epithelium of endometrium in the model group was lower than the blank group (P < 0.05). There were significant differences on the thickness of covering epithelium of endometrium in the high dose group and low dose group with the prescription of tonifying kidney and promoting blood circulation, the WZ group and the model group (P < 0.01). The sum of gland number and gland area as well as the maximum diameter / minimum diameter of the model group was significantly lower than that of the blank group (P< 0.05, orP < 0.01). The shape factor was the closest to “1”, which had significant difference compared with the blank group (P < 0.01). Compared with the model group, the sum of gland number and glandular lumen of the high dose and low dose group were increased at different levels (P < 0.05, orP < 0.01). The sum of gland number, glandular lumen area, shape factor, and maximum diameter / minimum diameter of WZ group had significant difference compared with the model group (P < 0.05, orP < 0.01). The sum of interstitial cells’ nucleus area, integral optical density of interstitial cells’ nucleus and number of interstitial blood vessels in the high dose and low dose group were significantly higher compared to the model group (P < 0.05, orP < 0.01). The sum of interstitial cells’ nucleus area and the integral optical density of interstitial cells’ nucleus in the WZ group were obviously higher than the model group (P < 0.01). It was concluded that the external method of tonifying kidney and promoting blood circulation can effectively improve the endometrial morphology of kidney-deficiency and blood-stasis rat model, promote the synchronous development of endometrial gland and stroma, in order to play a role to improve the endometrial receptivity.
5.Expression of hsa-miR-20a in human glioma tissues and its effect on the proliferation of human glioma cells in vitro.
Xingjun YAO ; Wei LI ; Yuehua WANG ; Zhengbin DING ; Wenzhong HOU ; Minmin ZENG ; Guangce DENG ; Jinzhu ZHANG ; Hongfa YANG
Journal of Southern Medical University 2012;32(2):198-201
OBJECTIVETo investigate miR-20a expression in human glioma and normal brain tissues and its effect on the proliferation of glioma cells in vitro.
METHODSThe expression of miR-20a was detected in human normal brain tissues and glioma tissues by real-time RT-PCR. miR-20a mimics were synthesized and transfected into U251 cells via liposome, and the cell proliferation were detected using MTT assay and flow cytometry.
RESULTSThe glioma tissues showed significantly up-regulated expression of miR-20a compared with normal brain tissues (P=0.035). The expression level of miR-20a was higher in high-grade than in low-grade gliomas. miR-20a mimics significantly enhanced the proliferation of U251 cells and the percentage of S-phase cells.
CONCLUSIONmiR-20a shows potent effect in promoting the growth of glioma cells, suggesting its important role in the pathogenesis of human glioma.
Adult ; Brain Neoplasms ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Female ; Glioma ; genetics ; metabolism ; pathology ; Humans ; Male ; MicroRNAs ; genetics ; metabolism ; Middle Aged ; Real-Time Polymerase Chain Reaction ; Young Adult
6.Failure Causes Analyses and Corrective Actions of Two Different Types of Bowie-Dick Test Pack
Riqing LIN ; Lihua DENG ; Yunfang MA ; Jinzhu LIU
Chinese Journal of Nosocomiology 2005;0(11):-
OBJECTIVE To analyze the monitoring performance and testing efficacy and reliability of standardized disposable B-D test pack and standard linen test pack,and summarize the corrective actions based on failure analyses for future reference.METHODS The two different test packs were employed to evaluate the vacuum performance of dynamic air removal sterilizer.RESULTS We enrolled 400 standardized disposable B-D test packs and standard linen test packs respectively.The disposable pack failed 4 cases with the success ratio 99%,while the linen pack failed 30 cases with the success ratio 92.5%.CONCLUSIONS The poor conformity of hand-made linen test pack,variation of steam pressure,sterilizer failure,and unprofessionalism of sterilizer operator contribute as main failure causes in B-D test.Standardized disposable B-D test pack can decrease subjective factors significantly,and the test results are more reliable and standardized.

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