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.Exploring the Integrated Traditional Chinese and Western Medicine Treatment for Acute Respiratory Distress Syndrome from the Perspective of State Differentiation and Treatment
Yikun GUO ; Miao CHENG ; Jun YAN ; Bei XUE ; Linwen CHEN ; Shujiao LI ; Shangshang JIANG ; Liangduo JIANG ; Chengjun BAN
Journal of Traditional Chinese Medicine 2024;65(6):577-581
From the perspective of state differentiation and treatment, it is believed that the pathogenesis of acute respiratory distress syndrome (ARDS) is that evil poisons injured the lungs, and the lung qi suddenly collapsed, then blocked and exhausted, and the qi failure to control blood and liquid, then the fluids overflow outside the vessels, and damp phlegm, stasis, and toxins became knotted up in the body, which ultimately leads to qi dysfunction, and a series of symptom arise, so qi impairment is the principal mechanism of ARDS. A combination of Chinese and Western medicine was proposed to treat ARDS by combining tangible qi and intangible qi, using Chinese herbal medicine to boost qi and relieve collapse, percolate and drain dampness with bland medicinals, resolve toxins and dissolve stasis, and regulate qi, and combining with Western medicine to assist qi circulation to improve qi's consolidation, propulsion, and transformation, so as to make the evil qi go away, the positive qi restored, the viscera qi circulated, qi, blood, yin, and yang connected, and the activities of life maintained, and thus to achieve the goal of treating ARDS by integrated Chinese medicine and Western medicine.
3.Value of blood routine indexes and their ratios in judging the prognosis of adult patients with extensive burns
Jianjun ZHENG ; Zi′en WANG ; Linwen ZHENG ; Zhaorong XU ; Shun CHEN ; Zhaohong CHEN
Chinese Journal of Burns 2020;36(12):1167-1172
Objective:To investigate the value of neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), and blood platelet count (BPC) in judging the prognosis of adult patients with extensive burns.Methods:From January 2012 to December 2018, 99 adult patients with extensive burns who met the inclusion criteria were admitted to Union Hospital of Fujian Medical University, including 76 males and 23 females, aged 18 to 75 (43±13) years. According to the prognosis, the patients were divided into survival group of 79 cases and death group of 20 cases. Their clinical data were retrospectively analyzed by the method of case-control study. The gender, age, total burn area, inhalation injury, use of mechanical ventilation and white blood cell count, neutrophil count, lymphocyte count, and BPC on post injury day (PID) 1, 3, and 7 were collected, and the NLR, PLR, difference value of BPC on PID 3 and PID 1 (ΔBPC3), difference value of NLR on PID 3 and PID 1 (ΔNLR3), difference value of PLR on PID 3 and PID 1 (ΔPLR3), difference value of BPC on PID 7 and PID 1 (ΔBPC7), difference value of NLR on PID 7 and PID 1 (ΔNLR7), difference value of PLR on PID 7 and PID 1 (ΔPLR7) of patients in the two groups were calculated. Data were statistically analyzed with Mann-Whitney U test, independent sample t test, chi-square test to screen the death-related factors of patients. Binary classification single factor and multifactor logistic regression analysis were used to analyze the death-related factors of patients. The receiver′s operating characteristic (ROC) curve of the independent risk factor of death of patients predicting the prognosis of adult patients with extensive burns was drawn, and the area under the curve, the optimal threshold and its sensitivity and specificity were calculated. Results:(1) There were statistically significant differences in total burn area and use of mechanical ventilation of patients between the two groups ( Z=-2.615, χ2=7.282, P<0.01). (2) On PID 1, there was statistically significant difference in NLR of patients between the two groups ( Z=-2.414, P<0.05). On PID 3, there were statistically significant differences in BPC and ΔNLR3 of patients between the two groups ( Z=-2.048, -2.780, P<0.05 or P<0.01). On PID 7, there were statistically significant differences in lymphocyte count, BPC, NLR, and ΔNLR7 of patients between the two groups ( Z=-2.248, -2.231, -2.641, -3.669, P<0.05 or P<0.01). (3) Binary classification single factor logistic regression analysis showed that the total burn area, mechanical ventilation, BPC and NLR on PID 7, and ΔNLR7 were related to death of patients (odds ratio=1.038, 0.193, 0.990, 1.086, 1.105, 95% confidence interval=1.010-1.067, 0.062-0.598, 0.982-0.998, 1.012-1.165, 1.037-1.178, P<0.05 or P<0.01). Binary classification multifactor logistic regression analysis showed that ΔNLR7 was the independent risk factor of death of adult patients with extensive burns (odds ratio=1.090, 95% confidence interval=1.008-1.178, P<0.05). (4) The optimal threshold of ROC curve of ΔNLR7 for predicting the prognostic death of 97 adult patients with extensive burns was -0.073 4. The sensitivity under the optimal threshold was 65.0%, and the specificity was 78.5%. The area under the ROC curve was 0.776 (95% confidence interval=0.650-0.882, P<0.01). Conclusions:Dynamic monitoring of NLR and BPC is of great significance to assist in judging the prognosis of adult patients with extensive burns. ΔNLR7 is an independent predictor of death in adult patients with extensive burns, while PLR can not predict the death of adult patients with extensive burns.
4. Clinical effects of artificial dermis combined with vacuum sealing drainage and autologous split-thickness skin graft in repair of scar contracture deformity after extensive burn
Shun CHEN ; Linwen ZHENG ; Wei LIU ; Zhaohong CHEN
Chinese Journal of Burns 2019;35(8):608-610
Objective:
To explore the clinical effects of artificial dermis combined with vacuum sealing drainage (VSD) and autologous split-thickness skin graft in repair of scar contracture deformity after extensive burn.
Methods:
A total of 8 patients with scar hyperplasia and contracture deformity on joint after healing of extensive burn were admitted to our hospital from August 2015 to August 2017. There were 5 females and 3 males aged 8 to 45 years with an average of 23 years. In the first stage operation, scar tissue on contracture site was removed, and the wound was covered by artificial dermis followed by continued VSD treatment. On 10-14 d after the first stage operation, the artificial dermis tissue formed, and the second stage autologous split-thickness skin grafting and continued VSD treatment were performed. Routine anti-scar therapy was carried out after healing of wounds. Time of wound healing after the second stage operation was recorded. Colour and texture of the split-thickness skin graft, scar formation condition of the donor site, and action condition of the operation site during follow-up were observed.
Results:
The wounds of 8 patients were healed in 10-14 d after the second stage operation. During follow-up of 6-24 months, the split-thickness skin graft was with smooth surface and good elasticity, and the function of joint recovered well. The donor site in head healed well with no scar. Only pigmentation was left in the donor site of thigh, and scar was not obvious. The patients and their family members were satisfied.
Conclusions
After application of artificial dermis combined with VSD and autologous split-thickness skin graft in repair of scar contracture deformity after extensive burn, the skin grafting area and donor site were with unobvious scar hyperplasia, and the joint function in the operation area was good.
5.Clinical study on the regimen of medicinal wine based on constitution distinction of TCM
Zhiyuan CHENG ; Suliu WU ; Xuzhi XU ; Chunfu DONG ; Linwen PAN ; Jianping CHEN ; Shuping PAN
International Journal of Traditional Chinese Medicine 2014;36(2):110-113
Objective To observe the effect of medicinal wine regimen which is guided by the theory of constitution distinction of TCM.Methods 76 patients were selected from all patients enrolled in wine style health experience from February 2011 to December 2011 in line with the corresponding physical characteristics.Among these 76 patients,23 patients were Yang deficiency constitution,34 were phlegm dampness constitution,and 19 were yin deficiency constitution.All these patients were taken a physical and chemical examination after taking the medicinal wine for a month.Results ①Examination index comparison before and after the treatment in different physical constitution patients:in phlegm dampness constitution patients the pulse were slowed (t=2.122,P<0.05) after taking the blood fat reducing wine,and physical type of conversion points were lowered (t=4.86,P<0.01); in Yang constitution patients,the systolic blood pressure (FT4),free thyroxine T4,physical type of conversion points were all significantly decreased [t values were 2.735,3.672,12.93,P<0.05 or 0.01] after taking Guilu Erxian wine; in Yin deficiency patients,adrenocorticotropic hormone (ACTH),interleukin-1 (IL-1),interleukin-2 (IL-2) were all increased [t values were 2.355,2.645,3.829,P<0.05 or 0.01] after taking Ziyin-Buxue wine.② Comparison of the clinical curative effect:according to the type of constitution transformation changes,the total effective rate was more than 70%.According to Ridit test,except yin deficiency physique (t=1.85,P>0.05),patients in both phlegm dampness constitution and yang deficiency constitution groups showed statistical difference after the treatment than before (t=4.86 and 12.93,P <0.01).Conclusions Blood fat reducing wine can improve phlegm dampness constitution; Ziyin-Buxue wine can improve Yin deficiency constitution and enhance immune function; Guilu Erxian wine can lower blood pressure and improve Yang deficiency constitution.

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