1.Multi-center Randomized Controlled Clinical Trial of Huangqi Injection Combined with Buzhong Yiqi Acupuncture in Treatment of Chronic Fatigue Syndrome with Qi Deficiency
Chengcheng WANG ; Xing TANG ; Chunmei LI ; Zhongbo WANG ; Yanlin FU ; Min DAI ; Min YANG ; Congcong YU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(7):163-169
ObjectiveTo investigate the clinical efficacy of Huangqi injection combined with Buzhong Yiqi acupuncture in the treatment of chronic fatigue syndrome (CFS) with Qi deficiency and its effects on TCM syndromes, fatigue symptoms, serum superoxide dismutase (SOD), malondialdehyde (MDA), and oxidized low-density lipoprotein (ox-LDL) levels. MethodA total of 200 patients with CFS of Qi deficiency were randomly divided into a control group (100 cases) and an observation group (100 cases). The control group was treated with vitamin B compounds, and the observation group was treated with Huangqi injection combined with Buzhong Yiqi acupuncture for two weeks. The scores of TCM syndromes, fatigue symptoms, levels of serum SOD, MDA, and ox-LDL and the incidence of adverse reactions were observed and compared before and after treatment in two groups. ResultAfter treatment, the total effective rate of the control group was 54.34% (50/92), while that of the observation group was 88.54% (85/96). The total effective rate of the observation group was higher than that of the control group (χ2=27.13,P<0.05). Compared with those in the two groups before treatment, scores of fatigue self-assessment scale (FSAS), physical fatigue and mental fatigue, and sleep/rest response scores of fatigue in the two groups after treatment were significantly decreased (P<0.05). After treatment, scores of FSAS, physical fatigue and mental fatigue, and sleep/rest response scores of fatigue in the observation group were significantly decreased compared with those in the control group (P<0.05). Compared with those in the two groups before treatment, TCM syndrome scores in the two groups after treatment were significantly decreased (P<0.05). After treatment, TCM syndrome scores in the observation group were significantly decreased compared with those in the control group (P<0.05). Compared with those in the two groups before treatment, MDA levels in the two groups were significantly decreased (P<0.05), ox-LDL levels in the observation group were significantly decreased (P<0.05), and SOD levels were significantly increased (P<0.05). After treatment, compared with those in the control group, the serum MDA and ox-LDL levels in the observation group were significantly decreased (P<0.05), and the serum SOD was significantly increased (P<0.05). No serious adverse events or adverse reactions occurred during this clinical trial. ConclusionHuangqi injection combined with Buzhong Yiqi acupuncture has a good clinical curative effect in the treatment of CFS with Qi deficiency, which can effectively improve the fatigue symptoms of patients, increase the level of SOD, and reduce the level of serum MDA and ox-LDL. It is related to the production of antioxidants, inhibiting the production of lipid peroxides, and improving the body's ability to resist oxidative stress.
2.Research progress of role and mechanism of ferroptosis in acute kidney injury
Jian WANG ; Wei JI ; Bingyu REN ; Zhongbo LI ; Qian HE
Journal of Clinical Medicine in Practice 2024;28(18):129-134
Ferroptosis is a novel strategy for regulating cell death, and its occurrence is closely related to intracellular iron metabolism, lipid metabolism, amino acid metabolism, and various signaling pathways. Ferroptosis plays a pivotal role in the pathogenesis of acute kidney injury (AKI). Ferroptosis may serve as an important target for the treatment of AKI caused by various reasons such as ischemia-reperfusion injury, nephrotoxic drugs, rhabdomyolysis syndrome, sepsis, and so on. This paper reviewed the regulatory mechanisms of ferroptosis and its research progress in AKI.
3.Preliminary value of multimodal MRI radiomics in the diagnosis of neonatal white matter lesions
Hong YU ; Jingshi WANG ; Zhongbo CHEN
Chinese Journal of Postgraduates of Medicine 2023;46(9):810-815
Objective:To investigate the effectiveness of multimodal magnetic resonance imaging (MRI)-based radiomics models in the auxiliary diagnosis of neonatal cerebral white matter lesion (WML).Methods:Clinical and MRI imaging data were collected from 91 children diagnosed with WML in Dalian Women and Children′s Medical Center from April 2018 to December 2021, including 58 cases in the good prognosis group and 33 cases in the poor prognosis group. The region of interest (ROI) was drawn for the lesion of each sequence image (T 1WI, T 2WI, DWI and SWI). Three models, T 1WI+T 2WI (model 1), T 1WI + T 2WI + DWI (model 2) and T 1WI + T 2WI + SWI (model 3), were created. Resample of the ROI, pre-processing of images and extraction of features were performed by using PyRadiomics, and the extracted features were standardized on the Dr.Wise research platform. A logistic regression classifier was used to create the radiomics model and 5-fold cross-validation was carried out 10 times. Key features were screened, and the receiver operating characteristic (ROC) curve was drawn to evaluate the predictive ability of the models. And DeLong was used to compare the effectiveness of different models. Results:The AUC values of model 2 (0.95) and model 3 (0.93) in the validation set were higher than that of model 1 (0.90). The accuracy, sensitivity and specificity of model 1, model 2, and model 3 were (86%, 89% and 87%), (79%, 88% and 88%) and (90%, 90% and 86%), respectively. The differences between model 1 and model 2 by DeLong′s test was statistically significant ( P<0.05), whereas there was no statistically significant differences between model 3 and model 1 or between model 3 and model 2 ( P>0.05). Conclusions:Multimodal MRI-based radiomics model was proved to be an effective tool in the auxiliary diagnosis of neonatal WML.
4.Effect of Banxia Xiexintang-containing Intestinal Absorption Solution on Migration and Invasion of PMN-MDSCs in Gastric Cancer Microenvironment
Jingjing WEI ; Zhongbo ZHU ; Xiping LIU ; Peiqing LI ; Qiming CHEN ; Lirong DAI ; Lijuan SHI ; Haijing DUAN ; Qingmiao WANG
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(10):48-57
ObjectiveTo observe the effect of Banxia Xiexintang containing intestinal absorption solution (BXCIAS) on migration and invasion of polymorphonuclear myeloid-derived suppressor cells (PMN-MDSCs) in gastric cancer microenvironment. MethodThe complex solution (containing 0.63 g·mL-1 crude drug) was prepared. Gastric cancer cells were subjected to non-contact co-culture with PMN-MDSCs in Transwell chamber to create gastric cancer microenvironment. Cell counting kit-8 (CCK-8) assay was used to screen the optimal intervention concentration and time of BXCIAS on PMN-MDSCs for subsequent experiment. The blank group, model group, FAK inhibitor group, and BXCIAS groups (26%, 18%, and 10%) were designed. Scratch assay and Transwell assay were employed to detect the migration and invasion ability of PMN-MDSCs, and enzyme-linked immunosorbent assay (ELISA) to measure the expression of vascular endothelial cell growth factor (VEGF) and matrix metalloproteinase-9 (MMP-9) in tumor microenvironment. The expression levels of PMN-MDSCs pathway-related proteins FAK, phosphorylated (p)-FAK, protein tyrosine kinase (Src), and p-Src were detected by Western blot. ResultThe inhibition rates of PMN-MDSCs by 5%, 50%, 75%, and 100% BXCIAS at 48 h were higher than those at 24 h (P<0.05, P<0.01). The inhibition rate of PMN-MDSCs by 50% BXCIAS at 72 h was lower than that at 48 h (P<0.01), and the inhibition rates by 5% and 100% BXCIAS at 72 h were higher than those at 48 h (P<0.05, P<0.01). There was no significant difference in the inhibition rate by other concentration levels at 48 h. The half-maximal inhibitory concentration (IC50) at 48 h was 18.09%, indicating that 18% BXCIAS and 48 h were the optimal concentration and time, respectively. The migration distance of PMN-MDSCs was large (P<0.01), and the number of migrating and invading cells increased (P<0.01) in the mode group compared with those in the blank group. Compared with model group, FAK inhibitor and BXCIAS at different concentration decreased the migration distance of PMN-MDSCs (P<0.01), and the number of migrating and invading cells (P<0.01), especially the 26% BXCIAS (P<0.01). The expression of PMN-MDSCs pathway-related proteins FAK, p-FAK, Src and p-Src (P<0.01) and the expression of VEGF and MMP-9 (P<0.01) were higher in the model group than in the blank group. Compared with model group, FAK inhibitor and BXCIAS (26%, 18%, 10%) decreased the expression of FAK, p-FAK, and Src (P<0.01), and FAK inhibitor and 18% BXCIAS reduced the expression of p-Src (P<0.01), and the expression of VEGF and MMP-9 (P<0.01). ConclusionBXCIAS can inhibit the migration and invasion of PMN-MDSCs by down-regulating the expression of FAK, p-FAK, Src, and p-Src proteins in the FAK signaling pathway of PMN-MDSCs in gastric cancer microenvironment.
5.Effect of Banxia Xiexintang-containing Intestinal Absorption Solution on PMN-MDSCs Apoptosis in Gastric Cancer Microenvironment
Jingjing WEI ; Zhongbo ZHU ; Xiping LIU ; Peiqing LI ; Qiming CHEN ; Lirong DAI ; Lijuan SHI ; Haijing DUAN ; Qingmiao WANG
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(10):58-64
ObjectiveTo observe the effect of Banxia Xiexintang (BXT)-containing intestinal absorption solution on the apoptosis of polymorphonuclear myeloid-derived suppressor cells (PMN-MDSCs) in gastric cancer microenvironment. MethodBXT-containing intestinal absorption solution was prepared, and gastric cancer cells and PMN-MDSCs were non-contact co-cultured in Transwell chamber to establish gastric cancer microenvironment. Cell counting kit-8 (CCK-8) assay was used to screen the optimal intervention concentration and time of 0-100% BXT-containing intestinal absorption solution prepared by 0.63 g·mL-1 reconstitution solution. Cells were classified into blank group, model group, oxaliplatin group (10 mg·L-1), and BXT (26%, 18%, 10% BXT-containing intestinal absorption solution) group, and the apoptosis of PMN-MDSCs was detected by flow cytometry. The expression of apoptosis-related B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax), and cysteine-aspartic acid protease-3 (Caspase-3) in PMN-MDSCs was detected by Western blot. ResultAfter treatment for 24 h and 48 h, the PMN-MDSCs-inhibiting rate was increased by 5%, 50%, 75%, and 100% BXT-containing intestinal absorption solution compared with that in the blank group (P<0.05, P<0.01). At 72 h, the PMN-MDSCs-inhibiting rate by 50% BXT-containing intestinal absorption solution was lower than that at 48 h (P<0.01), and the PMN-MDSCs-inhibiting rate by 5%, 75%, and 100% BXT-containing intestinal absorption solution showed no significant difference from that at 48 h. Moreover, the half-maximal inhibitory concentration (IC50) at 48 h was 18.40%. Thus, 18% BXT-containing intestinal absorption solution and 48 h were the optimal intervention concentration and time. The survival rate of PMN-MDSCs in model group was higher than that in the blank group (P<0.05), and the apoptosis rate was lower than that in the blank group (P<0.05). Compared with model group, BXT containing intestinal absorption solution lowered the survival rate and raised apoptosis rate of PMN-MDSCs (P<0.05), particularly the 26% BXT-containing intestinal absorption solution (P<0.05). The expression of Bax and Caspase-3 in PMN-MDSCs was lower in the model group than in the blank group (P<0.05), and the expression of Bcl-2 was higher in the model group than in the blank group (P<0.05). The expression of Caspase-3 in PMN-MDSCs increased (P<0.05) and the expression of Bcl-2 decreased (P<0.05) in oxaliplatin group and BXT group compared with those in the model group. The expression of Bax rose in oxaliplatin group and BXT group (10% BXT-containing intestinal absorption solution) (P<0.05). ConclusionBXT can induce the apoptosis of PMN-MDSCs by regulating the expression of apoptosis-related proteins Bax, Caspase-3, and Bcl-2 in gastric cancer microenvironment.
6.Factors affecting long-term survival of advanced high-grade serous ovarian cancer
Yuanming SHEN ; Liqin JIN ; Sangsang TANG ; Yu WANG ; Weiguo LYU ; Zhongbo CHEN ; Xing XIE
Chinese Journal of Obstetrics and Gynecology 2021;56(6):393-400
Objective:To identify the factors associated with long-term survival and guide the decision for primary surgery in patients with advanced high-grade serous ovarian cancer(HGSOC).Methods:In this case-control study, clinical parameters, including surgical and non-surgical associated factors, were collected and compared between the patients with short-term (<2 years) and long-term (>5 years) survival who all underwent primary debulking surgery (PDS) followed by carboplatin and paclitaxel chemotherapy from January 2004 to December 2016. Univariate analysis was examined by chi-square test and multivariate analysis was performed by logistic regression analysis.Results:There were 95 cases long-term survival (LTS group) and 77 cases short-term survival (STS group) in 698 newly diagnosed HGSOC patients with International Federation of Gynecology and Obstetrics (FIGO) stage Ⅲc and Ⅳ who met include and exclude criteria. (1) Univariate analysis showed that the proportion of complete cytoreduction with no visible residual disease (R0) at PDS and platinum sensitivity in LTS group were significantly higher than those in STS group ( P<0.01). The surgical complexity score (SCS), the preoperative serum CA 125 level and the ascites volume in the LTS group were significantly lower than those of the STS group (all P<0.05). In the LTS group, the preoperative incidence of lesions in retrograde peritoneum of the bladder, serosal and mesangial membrane of the small intestine, upper abdominal peritoneum and liver parenchyma were significantly lower than those in the STS group (all P<0.05). Multivariate logistic regression analysis showed that platinum sensitivity ( OR=0.016, 95% CI: 0.004-0.063, P<0.01), ascites volume >500 ml ( OR=3.193, 95% CI: 1.285-7.930, P=0.012), and SCS ≥8 ( OR=17.433, 95% CI: 2.281-133.25, P=0.003) were independent factors affecting long-term survival ( P>0.05). (2) Totally 37 of 95 in long-term survival and 16 of 77 in short-term survival achieved R0 cytoreduction at PDS. Univariate analysis showed that preoperative serum CA 125 level, preoperative lesion score, preoperative lesion (DS) score, ascites volume, platinum sensitivity,and SCS were significantly correlated with the R0 PDS (all P<0.05). Multivariate analysis showed that ascites volume >500 ml ( OR=5.199, 95% CI: 2.015-13.409, P=0.001), DS >2 ( OR=15.264, 95% CI: 5.843-39.874, P<0.01) and SCS ≥4 ( OR=4.176, 95% CI: 1.618-10.777, P=0.003) were independent factors associated with R0 cytoreduction. In patients with DS ≤2 or SCS <4, but not those with DS >2 or SCS ≥4, R0 cytoreduction was significantly associated with long-term survival. Conclusion:The intrinsic biology of tumor is the factor influencing long-term survival of advanced HGSOC patients, and those who present with wide intraperitoneal metastases and need to remove multiple organs may not benefit from R0 cytoreduction.
7.Performance evaluation of Sysmex XN-9000 automated hematology analyzer
Zhongbo XIONG ; Xiaoyan JIN ; Bo LU ; Lei WANG
International Journal of Laboratory Medicine 2015;(10):1373-1375
Objective To evaluate the performance of Sysmex XN‐9000 automated hematology analyzer .Methods According to international and domestic standards ,performance of analyzer was evaluated .Results The within‐batch and between‐batch preci‐sion ,carryover pollution rate ,linearity range and the accuracy of Sysmex XN‐9000 analyzer were all conform to related require‐ments .Leukocyte classification results compared with manual classification ,the correlation of neutrophil ,lymphocyte ,monocyte and eosinophil were fine ,but correlation of basophil was not very ideal .Conclusion The performance of Sysmex XN‐9000 analyzer could be satisfying ,could meet the needs of clinical inspection and diagnosis and treatment .
8.The characteristics of enzyme kinetics of CTX-M-14 type extended-spectrum β-lactamase with Pro167 residue substitution
Rong XU ; Zhongbo SHANG ; Junwei HUANG ; Dongqing HAN ; Zhen WANG ; Yongliang LOU ; Xiushu CHEN
Chinese Journal of Microbiology and Immunology 2011;(3):250-254
Objective To analyze and evaluate the characteristics of enzyme kinetics of CTX-M-14 type extended-spectrum β-lactamase(ESBL) with Pro167 residue substitution. Methods By molecular cloning and PCR techniques, CTX-M-14 gene was directionally cloned into plasmid pET28a( + ) from a clinical E. coli isolate and formed an expression vector to transform competent E. coli BL21 (DE3 ). Prol67 residue substitutions of P167G, P167Q, P167S and P167T were introduced to CTX-M-14 by site-directed muta-genesis based on overlap extension PCR with the former recombinant plasmid as PCR template, respectively.The wild-type CTX-M-14, recombinant CTX-M-14 protein and its variants were expressed and purified, then their steady-state kinetic parameters (Kcat, Km and Kcat/Km ) against β-lactam antibiotics were determined.Results The kinetic parameters of wild-type and recombinant CTX-M-14 had no statistically significant differences (P>0.1). The 1/Km, Kcat and Kcat/Km values of P167S variant against ceftazidime were 16-fold, 2.87-fold and 43.6-fold higher than those of recombinant CTX-M-14, respectively, and the Kcat/Km value of P167S variant against penicillin, ampicillin, cefazolin, cefuroxime, ceftriaxone, cefotaxime decreased( < 0.05). Compared with the kinetic parameters of recombinant CTX-M-14, the kinetic parameters of P167T variant against ceftazidime had no significant change, but the Kcat values of P167Q and P167G variants decreased dramatically(P<0.01). Conclusion There was no difference between the enzyme activities of wild-type and recombinant CTX-M-14. P167S variant could not only promote the enzyme affinity of CTX-M-14 to ceftazidime but also improve the conversion rate of enzyme-substrate complex in the ceftazidime hydrolysis. The comparison of the kinetic parameters of CTX-M-14 and its variants with Pro167 residue substitution showed that the increased activity of CTX-M ESBL variants against ceftazidime could not be simply explained with the enlarged cavity in active site that may be caused by the replacement of Pro167 residue by smaller amino acids.
9.Effect of exogenesis vascular endothelial growth factor gene on the apoptosis of brain cells in the HIBD of newborn rats
Hong JI ; Lixin XU ; Yunxia QU ; Zhongbo WANG
Chinese Pediatric Emergency Medicine 2010;17(3):-
Objective To explore the effect of exogenesis VEGF 120 gene on the apoptosis of brain cells in the HIBD of newborn rats. Methods VEGF eukaryotic expression plasmid (pCDNA 3.1/r VEGF 120) was constructed by cloning rat VEGF 120 cDNA into eukaryotic expression vector pCDNA 3.1. The HIBD model was established with seven days old SD rats,and all rats were diveded into two groups at random :contral group 18 rats( every rat model was injected pCDNA 3.1 100 μg immediately after hypoxia-is-chemic.then raised seven days) and therapy group 18 rats (every rat model was injected pCDNA 3.1/ rVEGF 120 100 μg immediately after hypoxia-ischemic). Flow cytometer( FCM) was used to detect the ratio of apoptosis of brain cell. Results There was a significant decrease in the ratio of apoptosis brain cells( control group 17.505 ± 0.949; therapy group 8.93 ± 0. 332). Conclusion The VEGF gene product can reduce apoptosis of brain cells.
10.Runx2 is Involved in Regulating Osterix Promoter Activity and Gene Expression
Dongmei SUN ; Zhongbo LIU ; Yan ZHAO ; Zhenwei GONG ; Dan LI ; Xiyuan WANG ; Xianlu ZENG ; Wenguang LIU
Progress in Biochemistry and Biophysics 2006;33(10):957-964
Though Runx2 and Osterix are both key transcription factors in the pathway of osteoblast differentiation, whether Runx2 positively regulates Osterix being unknown. It was showed that Runx2 induced the gene expression of Osterix both in the non-osteoblastic cell lines, either pluripotent or differentiated, and in the osteoblastic cell lines. At the same time, the results also indicated that Runx2 up-regulated the activity of the 3.2 kb human Osterix promoter. Further experiments identified a highly conserved and functional Runx2 binding site "AGTGGTT" within the promoter. Thus the results support the hypothesis that Runx2 is involved in the regulation of the Osterix gene expression. Moreover, the transient transfection and dual-luciferase assay showed Osterix up-regulated the activity of the 2.3 kb type Ⅰ collagen promoter in the non-osteoblastic cells, but Runx2 did not. This difference implies that Osterix, the down stream transcription factor of Runx2 during osteoblast differentiation, is needed to stimulate the osteoblast-specific gene expression of type Ⅰ collagen.


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