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.Effects of body mass index, appendicular skeletal muscle mass index and serum lipid levels on the risk of tumor progression in patients with high-risk renal clear cell carcinoma
Danping ZHENG ; Yancai LIANG ; Zhiyuan ZHANG ; Jian CUI ; Jingxiao HAO ; Xiangyun LU ; Juan WANG ; Na GUO ; Kang YU
Chinese Journal of Clinical Nutrition 2022;30(4):199-205
Objective:To evaluate the potential effects of serum lipid levels, appendicular skeletal muscle mass index (ASMI) and body mass index (BMI), together with its dynamic changes, on tumor progression in renal clear cell carcinoma patients, so as to inform body weight management.Methods:This prospective cohort study included a total of 100 patients with high-risk clear cell renal cell carcinoma. Serum lipid levels were detected, ASMI and BMI were measured using bioelectrical impedance analysis and the dynamic changes of BMI were tracked. The effects of BMI, ASMI and serum lipid levels on tumor progression within 2 years were explored.Results:Patients with normal BMI and low ASMI had 5.248 (95% CI: 1.946 to 14.153, P = 0.001) times higher risk of tumor progression than those who were overweight or obese. For every 0.1-unit increase in pre-operative HDL-C, the risk of tumor progression decreased by 0.771 (95% CI: 0.631 to 0.942, P = 0.011) times. Patients who experienced more than 5% decrease in BMI compared with baseline had 5.165 (95% CI: 1.735 to 15.370, P = 0.003) times the progression risk of patients whose BMI changed within ±5% from baseline. Conclusions:The advantage of obese clear cell carcinoma patients over normal-weight patients in tumor progression-free survival may be influenced by ASMI, pre-onset involuntary weight loss and lipid levels. Therefore, patient weight management should not merely focus on absolute BMI but tailor to individual characteristics, including cancer stage, body composition and metabolic status.
3.Performance verification and evaluation of 4 domestic chemiluminescence systems on 8 tumor mark-ers
Yan SHI ; Yancai WEI ; Weiling ZHENG ; Jialing WEI ; Miaoli SONG ; Gengchao ZHU ; Xun LU ; Chen YANG
Chinese Journal of Nuclear Medicine and Molecular Imaging 2018;38(12):801-804
Objective To validate the performance of 4 domestic chemiluminescence immunoassay (CLIA) systems on 8 tumor markers quantitative assay kits. Methods Four domestic CLIA systems were randomly marked as A, B, C, D and 8 tumor markers, including carbohydrate antigen (CA)125, CA15-3, CA19-9, ferritin (Fer), alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA), prostate-specific an-tigen (PSA) and free PSA (fPSA) were determined. According to the standard of Clinical and Laboratory Standards Institute (CLSI), the precision, methodological comparison and analytical measure range of 4 systems were validated. Clinical serum samples were obtained from patients in Suzhou Hospital. According to the CLSI EP9-A3 protocol, imported equipment was used as the reference system. The biases of medical de-cision points were assumed, and Pearson correlation analysis and Spearman correlation analysis were used to analyze the data. Results The precision verification of CA125 and PSA on A, CA125 and AFP on B, CA125, CEA, AFP and PSA on C, and all 8 tumor markers on D could meet the laboratory quality control requirements. The correlations of the test results between A-D and the imported equipment were significant (all P<0.05) with the correlation coefficients 0.79-0.99, 0.47-0.99, 0.90-0.98 and 0.78-1.00, respec-tively, and the number of acceptable tests at the level of medical decision was 5, 2, 5, 4. All tests were certified to meet the analytical measure range validation. Conclusions The detection performance of 4 do-mestic CLIA systems for all 8 tumor markers are different. The performance of domestic CLIA systems should be tested when choosing one that can meet laboratory quality control requirements.
4.Clinical study on bacterial condition in amniotic cavity in the third trimester of pregnancy
Yinghua ZHAO ; Jieyun HE ; Yancai ZHENG ; Bing LIN ; Xiuqun ZHANG ; Zhuoshu ZHAO ; Yili TAN
Clinical Medicine of China 2011;27(1):100-103
Objective To study bacterial conditions in amniotic cavity of the third trimester and the delivery. Methods Patients underwent cesarean section due to various reasons were randomly enrolled in the study. Ten ml aqua amnii taken from using aspesis were cultured and the bacteria were identified and tested for drug sensitivity. All patients were not treated with antibiotics before and during operation,conventional antibiotics treatnent was used after operation. The patients were divided into 3 groups based on their status: 34 cases of premature rupture of membrane ( PROM ), thirty-four cases of labor without PROM, and 27 cases of pregnancy without PROM. The positive rate, species , distribution and drug sensitivity of bacteria in the 3 groups were analyzed. Results The positive rate was 61.8% (21/34) in PROM ,52. 9% ( 18/34 ) in labor without PROM,11.1% (3/27)in pregnancy without PROM. The positive rates were significantly different among the 3 groups (x2 = 17.29 ,P =0.000). However, there was no significant difference between the labor without PROM group and the pregnancy without PROM group ( x2 = 0.541 , P = 0.462 ). Staphylococcus epidermidis was the most common isolated bacteria, following by staphylococcus aureas, streptococcus, colon bacillus, and pseudomonas. There were no significant differences of bacterial species between labor without PROM and pregnancy without PROM( x2 = 11.9,P =0.535 ). The relative higher positive rate in the labor without PROM and PROM indicated that they were important inducement of bacteria infection. There were no significant difference on positive rate and bacteria species distribution between the PROM and labor without PROM group,which suggested that up-bound along vagina to amniotic cavity was the most common route of infection. Sensitive antibiotics should be given to the patients of PROM and labor without PROM targeted at staphylococcus,streptococcus, colon bacillus and monad. Antibiotics such as Cephems, Penicillins and Clindamycin were sensitive to various bacteria and safe to both mother and baby, thus were recommended. Quinupristin, Vancomycin,Furantoin, Fusidic ,Teicoplanin, Amikacin and Meropenem were also sensitive to various bacteria, but due to their side effects and the principle of ladder antibiotics using, they were recommended as second line antibiotics. Conclutions Up-bound along vagina to amniotic cavity is the most common route of infection. For rupture of membrane and labor without PROM patients, antibiotics should be given, and strict aseptic technique,washing cavity and incision, using antibiotics after operation are necessary to prevent infection.
5.Clinical effect of oxaliplatin-containing and cisplatin -containing regimen in TACE of advanced hepatocellular carcinoma
Yingqiang JIANG ; Yancai CHEN ; Zongwan CHEN ; Lianxi ZHENG ; Ke YANG ; Lauqiong GOU ; Xu LIN ; Ping HE
Cancer Research and Clinic 2009;21(10):687-689
Objective To explore the clinical effect and side-effect of oxaliplatin(L-OHP)-containing regimen and cisplatin (DDP)-containing regimen in TACE of advanced hepatocellular carcinoma (HCC). Methods 108 patients with advanced HCC were randomly divided into experimental group(n=55) and control group (n =53). The experimental group were treated with TACE using L-OHP. After dilute with glucose solution, L-OHP(130 mg/m2) and FT207 (500-750 mg/m2) were injected into blood vessel respectively. ADM (40 mg/m2) and LP (10~30 ml) were emulsified and then used for vessel embolism according to the size of focus. The control group received TACE with DDP, DDP (40 mg/m2) and F]'207(500~750 mg,/m2) were diluted with glucose solution, and also according to the size of tumor' s focus, ADM(40 mg/m2) and LP(10~30 ml) were emulsified for vessel embolism, and then diuretic. Results The total effective rate of experimental group was 67.3 % (37/55), and that of control group was 47.2 % (25/53), and the difference was with statistical significance (P <0.05). The descent rate of AFP of experimental group was 73.1%(31/43), and that of control group was 44.7 % (17/38), and the difference was with statistical significance (P <0.05). The main side effects were gastrointestinal reactions, the incidence rate of nausea and vomiting in experimental group were lower than control group, and the difference was with statistical significance. The incidence rate of leucopenia, damage of hepatic function and peripheral neuritis were not significant. Damage of heart and kidney were not found in the two sets. Conclusion L-OHP -containing regimen in TACE of advanced HCC is an efficient method, with good security and good tolerance to patients.

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