1.Analysis of Alleviating Effect of Calcium Cyanamide on Replanting Problems of Rehmannia glutinosa
Lianghua LIN ; Hengrui ZHANG ; Haoxiang YU ; Fan YANG ; Yufei WANG ; Caixia XIE ; Tao GUO ; Zhongyi ZHANG ; Liuji ZHANG ; Bao ZHANG ; Suiqing CHEN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(22):212-222
ObjectiveTo investigate the alleviating effect of calcium cyanamide (CaCN2) soil fumigation on replanting problems of Rehmannia glutinosa. MethodsNewly soil (NP) was used as the control group, while three treatment groups were established: replanted soil (RP), newly soil treated with CaCN2 (120 g·m², tillage depth 25 cm) (NPCC), and replanted soil treated with CaCN2 (RPCC). R. glutinosa was cultivated in all groups. At harvest, the tuber agronomic traits (number of enlarged roots, maximum root diameter, fresh weight, dry weight) were measured. The content of catalpol and rehmannioside D was quantified by ultra-high-performance liquid chromatography (UPLC) to evaluate medicinal quality. Rhizosphere soil available nutrients and enzyme activities were analyzed by assay kits. The community structure and composition of fungi and bacteria in rhizosphere soil were assessed via internal transcribed spacer 2 (ITS2) sequencing and 16S rDNA sequencing, respectively. ResultsCompared with NP, the RP group showed obviously reduced in tuber agronomic traits and quality indicators (P0.05). However, the RPCC group showed significant improvement in agronomic traits and a notable increase in rehmannioside D content compared to RP (P0.05). The contents of available phosphorus and potassium in RPCC and NP groups were obviously lower than those in RP (P0.05). The polyphenol oxidase soil (S-PPO) activity in RP was obviously lower than in NP (P0.05), while sucrose soil (S-SC), acid phosphatase soil (S-ACP), and S-PPO activities in RPCC were obviously higher than in RP (P0.05). Microbial richness and diversity in RP were obviously higher than in NP (P0.05), whereas no significant differences were observed between the RPCC and NP. The relative abundances of fungal genera Nectria, Myrothecium, Tomentella, and bacterial genus Skermanella were obviousl lower in RPCC and NP than in RP (P0.05). Correlation analysis that S-ACP activity was positively correlated with the content of rehmannioside D (P0.05). Fungal genera Engyodontium and Alternaria, and bacterial genera Pir4 lineage, Pirellula, Methyloversatilis, Brevundimonas, Ralstonia, and Acidibacter were obviously positively correlated with tuber dry weight (P0.05). Conversely, fungal genera Pseudaleuria, Nectria, Haematonectria, Ceratobasidium, and bacterial genera Streptomyces, Skermanella, RB41, Gemmatimonas, and Bacillus were obviously negatively correlated with dry weight (P0.05). The fungal genus Alternaria and bacterial genera Brevundimonas, Ralstonia, Acidibacter, and Dongia showed positive correlations with medicinal quality of R.glutinosa tuber, while fungal genera Pseudaleuria, Nectria, Stachybotrys, Fusarium, Gibberella, Ceratobasidium, and bacterial genera Sphingomonas, Skermanella, RB41, Gemmatimonas, and Bacillus were obviously negatively correlated (P0.05). ConclusionCaCN2 soil fumigation can significantly improve enzyme activities in replanted Rehmannia rhizosphere soil, enhance the utilization of available nutrients, reshape microbial community structure of replanted R.glutinosa at the family and genus level, and notably improve tuber agronomic traits and medicinal quality. This study provides a novel approach to alleviating replanting problems and offers insights for the integrated development of standardized cultivation techniques, including soil disinfection, nutrient-targeted regulation, and microbial inoculant application.
2.Structure and Function of Rhizomicrobes Recruited by Acteoside in Root Exudates of Rehmannia glutinosa
Yongxiang ZHANG ; Bao ZHANG ; Lianghua LIN ; Fan YANG ; Shujuan XUE ; Li GU ; Zhongyi ZHANG ; Liuji ZHANG ; Suiqing CHEN
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(9):156-165
ObjectiveTo reveal the correlation of Rehmannia glutinosa-soil feedback process with the formation of its continuous cropping obstacles through the identification of the root exudates of R. glutinosa and analysis of the specific rhizomicrobes recruited by the root exudate. MethodThe root exudates of R. glutinosa seedlings germinated under sterilized condition and those enriched in the rhizosphere of R. glutinosa cultivated in the field were collected and analyzed using the ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS). The highly abundant compounds identified in the root exudates were added into blank soil, and the soil microbial community was profiled using Illumina Miseq sequencing. The bacterial and fungal functions were predicted by PICRUSt and FUNGuild, respectively. ResultThe identification results showed that seven phenylethanoid glycosides were found in R. glutinosa root exudates, and acteoside possessed the highest abundance. In the soil enriched with acteoside, the bacterial genera such as Agromyces, Pseudomonas, Lysobacter, Sphingobium, Pseudoxanthomonas and Sphingomonas were enriched. For the fungi, the genera Neocosmospora, Plectosphaerella and Dactylonectria, and the species such as Neocosmospora rubicola, Plectosphaerella cucumerina, Dactylonectria alcacerensis and Fusarium solani showed higher abundance. The functional analysis indicated the above-mentioned bacterial genera may realize rapid proliferation by utilizing, biodegrading and transforming phenylethanoid glycosides, and some potential fungal pathogens were colonized. ConclusionThe R. glutinsoa-soil feedbacks were likely generated by the phenylethanoid glycosides in the root exudates together with the specific rhizomicrobes. The investigations of R. glutinsoa-soil feedbacks under continuous cropping system are critical to the further understanding of the underlying mechanisms related to its continuous cropping obstacles.
3.Study on the mutation characteristics of streptomycin genes of Multidrug-Resistant Mycobacterium Tuber-culosis in Jiangxi Province
Shengming YU ; Lianghua XIA ; Jiahuan ZHAN ; Siqi LIU ; Wei WANG ; Liang YAN ; Kaisen CHEN
The Journal of Practical Medicine 2024;40(1):91-96
Objective To understand the molecular characteristics of Streptomycin(SM)resistance in multidrug-resistant tuberculosis(MDR-TB)in Jiangxi Province,and to explore the relationship between SM resistant genes(rpsL,rrs and gidB)mutations and SM resistant phenotypes in Beijing genotype TB.Methods 106 non-replicated MDR-TB isolates were collected from Gaoxin Branch of The First Affiliated Hospital of Nanchang University and Jiangxi Provincial Chest Hospital from January to December 2021,and tested for drug-resistance phenotypes,whether they were Beijing genotype or not and the characteristics of rpsL,rrs and gidB gene mutations.Chi-square test was performed to determine whether rpsL,rrs and gidB mutations were related to genotypes and drug-resistance phenotypes.Results Among 106 cases of MDR-TB,76 cases were resistant to SM.A total of 58 cases had rpsL 43A>G mutation,8 cases had 88A>G mutation,5 cases had rrs mutation,and 3 cases had gidB mutation.Statistical analysis showed that the coincidence rate of gene mutation and phenotypic drug-resistance detection was 89.6%,and the specificity and sensitivity were 86.7%and 90.8%,respectively.The isolated rate of Beijing genotype TB was 88.7%,and the drug-resistant gene mutations were mainly concentrated in rpsL and rrs,while the drug-resistant mutations of non-Beijing genotype were mainly concentrated in gidB;in addition,Beijing genotype bacteria were more prone to gene mutations(P = 0.013),but there was no difference in phenotypic drug-resistance.Conclusions Mutations in rpsL,rrs,and gidB genes have a good coincidence rate with phenotypic drug-resistance,and molecular biology can be used to detect directly drug-resistance genes to predict bacterial resistance;TB genotypes are strongly associated with streptomycin resistance characteristics.
4.Characteristics and related clinical factors of myocardial 68Ga-FAPI-04 uptake in patients treated with anti-tumor therapy
Xiaofeng YU ; Yaqian ZHOU ; Qianyun WU ; Yinyan ZHU ; Lian XU ; Lianghua LI ; Cheng WANG ; Jianjun LIU ; Yumei CHEN
Chinese Journal of Nuclear Medicine and Molecular Imaging 2024;44(11):650-654
Objective:To analyze the value of 68Ga-fibroblast activation protein inhibitor (FAPI)-04 PET/CT in the detection of myocardial injury in patients treated with anti-tumor therapy. Methods:A retrospective study was conducted on 164 patients who underwent 68Ga-FAPI-04 PET/CT to evaluate the efficacy of anti-tumor therapy in Renji Hospital, School of Medicine, Shanghai Jiao Tong University between August 2021 and March 2024. The patients were divided into 68Ga-FAPI-04-positive group ( n=63, 36 males, 27 females, age (66.7±9.6) years) and 68Ga-FAPI-04-negative group ( n=101, 42 males, 59 females, age (55.2±14.1) years) based on the uptake of left ventricular myocardium (LVM). Moreover, FAPI-04 uptake was analyzed based on different types and locations, and the corresponding SUV max differences were analyzed by Kruskal-Wallis rank sum test. The differences of SUV max between 68Ga-FAPI-04-positive group and 68Ga-FAPI-04-negative group were analyzed by Mann-Whitney U test. The clinical factors such as gender, age, body mass index (BMI), previous history of coronary heart disease, left ventricular ejection fraction (LVEF), smoking history, hypertension, diabetes, cancer types and immune checkpoint inhibitors (ICIs) treatment were collected, and their predictive values for LVM 68Ga-FAPI-04 uptake were investigated by the binary logistic regression analysis. Results:Fifty patients of the 68Ga-FAPI-04-positive group (79.4%, 50/63) showed focal uptake of LVM, 7 patients (11.1%, 7/63) showed multifocal myocardial uptake, and 6 patients (9.5%, 6/63) showed diffuse myocardial uptake. A total of 127 uptake lesions were found, and most of them were located in the septum (37.8%, 48/127). The SUV max of LVM in 68Ga-FAPI-04-positive group and 68Ga-FAPI-04-negative group were 4.00(3.10, 5.40) and 1.31(1.20, 1.40) respectively ( z=-10.82, P<0.001). Differences of the SUV max among focal uptake group, multifocal myocardial uptake group, and diffuse myocardial uptake group were not significantly different (4.00(3.00, 5.10) vs 7.60(3.60, 9.30) vs 3.95(3.05, 5.05); H=3.81, P=0.149). There is no statistically significant difference either in FAPI uptake among different sites of LVM ( H=1.51, P=0.825). Age, previous history of coronary heart disease, BMI, LVEF and ICIs treatment were independent predictive factors for positive 68Ga-FAPI-04 uptake in the LVM (odds ratio ( OR) values: 0.87-10.43, all P<0.05). Conclusion:68Ga-FAPI-04 PET/CT is a potential new imaging method for the visualization of myocardial injury in patients with anti-tumor therapy.
5.Kinetic metrics of 68Ga-FAPI-04 in patients with pancreatic cancer identified by total-body PET/CT dynamic imaging
Ruohua CHEN ; Xinlan YANG ; Yanmiao HUO ; Yining WANG ; Xiaofeng YU ; Ling Yee NG ; Chaojie ZHENG ; Haitao ZHAO ; Lianghua LI ; Yun ZHOU ; Gang HUANG ; Jianjun LIU
Chinese Journal of Nuclear Medicine and Molecular Imaging 2022;42(12):729-733
Objective:To investigate the kinetic metrics of 68Ga-fibroblast activation protein inhibitor (FAPI)-04 in pancreatic cancers and normal organs by using total-body PET dynamic imaging. Methods:From December 2020 to December 2021, 68Ga-FAPI-04 total-body PET/CT dynamic imaging were performed on 6 pancreatic cancer patients (3 males, 3 females, median age 55.5 years) in Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University. Images were respectively analyzed. Manual delineations of volume of interests (VOIs) on multiple normal organs and pathological lesions were performed and time-to-activity curves (TACs) were generated. A reversible two-tissue compartment model (2TCM) was fitted for each tissue TAC. Rate constants including K1, k2, k3 and k4, and the total volume of distribution ( Vt) were obtained and compared by tissue types. Wilcoxon rank sum test and Spearman correlation analysis were used for data analysis. Results:Kinetic metrics varied significantly among normal organs and pancreatic cancer lesions ( z values: 2.00-1 240.00, all P<0.05). The highest K1 among lesions was observed in primary tumor (0.30 min -1), which was observed in the spleen (1.42 min -1) among normal organs. The highest k2 among lesions was observed in peritoneal metastases (0.24 min -1), which was observed in the spleen (2.59 min -1) among normal organs. Primary tumor showed the highest k3 of 0.17 min -1 among lesions, and the pancreas had the highest k3 of 0.16 min -1 among normal organs. Primary tumor had the highest k4 of 0.03 min -1 among lesions, and the heart, lungs, parotid glands had high k4(0.06 min -1) among normal organs. Vt were higher in pathological lesions compared to normal organs, with the highest in primary tumor (13.78 ml/cm 3). There were correlations between Vt in lesions and SUV mean( rs=0.86, P<0.001) or SUV max ( rs=0.77, P<0.001). Conclusion:The rate constants including K1, k2, k3 and k4, and Vt of 68Ga-FAPI-04 vary among normal organs and lesions.
6.Preclinical evaluation of 89Zr labeled Daratumumab for imaging diagnosis of multiple myeloma
Haitao ZHAO ; Lianghua LI ; Weijun WEI ; Yumei CHEN ; Chun LYU ; Cheng WANG ; Jianjun LIU
Chinese Journal of Nuclear Medicine and Molecular Imaging 2022;42(2):68-73
Objective:To prepare 89Zr labeled Daratumumab and evaluate its feasibility in the imaging diagnosis of multiple myeloma (MM). Methods:According to the principle of 89Y (p, n) 89Zr nuclear reaction, 89Zr was produced by cyclotron solid target system (30 μA, 1.5 h) and automatic purification module. The radionuclide purity, half-life and impurity metal ion concentration were detected. Desferrioxamine (DFO) was coupled with Daratumumab and then chelated with 89Zr to prepare 89Zr-DFO-Daratumumab. The quality control analyses of three consecutive batches were carried out. Pharmacokinetic evaluation and 89Zr-DFO-Daratumumab microPET/CT imaging were performed in normal rabbits and orthotopic myeloma mouse models, respectively. The SUV in situ myeloma and that in normal bone were compared by independent-sample t test. Results:About 560 MBq of 89Zr was obtained, and there were only two characteristic energy peaks of 89Zr (909 keV and 511 keV) by γ spectrometer. The half-life of 89Zr was 78.2 h, and the content of metal impurities was small. 89Zr-DFO-Daratumumab was prepared with pH of 7.2, radiochemical purity of more than 99%, good stability in vitro, and sterility and endotoxin tests were passed. Pharmacokinetic studies in rabbits showed that 89Zr-DFO-Daratumumab was gradually distributed from blood to liver, spleen, kidney and bone joints over time and metabolism. The results of microPET/CT imaging in orthotopic myeloma mouse models showed that the SUVs of 89Zr-DFO-daratumumab in situ myeloma were significantly higher than those in normal bone (2 h: 0.22±0.02 vs 0.06±0.00; 1 d: 0.38±0.01 vs 0.08±0.00; t values: 8.89, 21.90, both P=0.001). Conclusion:89Zr and 89Zr-DFO-daratumumab are successfully prepared, and relevant quality control and biological evaluation in vivo and in vitro are completed, which verify the feasibility of 89Zr-DFO-Daratumumab in the imaging diagnosis of MM, thus laying a foundation for clinical transformation.
7.Analysis of prescription medications in elderly patients with cardiovascular diseases
Chinese Journal of Geriatrics 2018;37(10):1082-1084
Objective To analyze the clinical medications in elderly patients with cardiovascular diseases.Methods The medical records were extracted and statistically analyzed in 386 elderly patients with cardiovascular diseases in Department of Vasculocardiology of our hospital.Results The primary hypertension,coronary heart disease,and arrhythmias were the most common cardiovascular diseases in elderly patients.The most frequently prescribed medications were calcium channel blockers,angiotensin-converting enzyme inhibitors,and traditional Chinese medicine compound preparations,which accounted for 28.2%,21.5% and 19.4%,respectively.The drug combination therapy is often used in clinical practice,and the proportion of four,five,≥ six drugs combination therapy was 16.6%,21.2% and 29.0%,respectively.Conclusions It is important to pay more attention to reducing adverse reactions and improving the rationality and safety of the drugs combination therapy in the treatment of senile cardiovascular diseases.
8. Effects of allogeneic bone marrow mesenchymal stem cells on polarization of peritoneal macrophages in rats with sepsis
Yuanhua ZHENG ; Bing XIONG ; Yiyu DENG ; Wen LAI ; Shaoyi ZHENG ; Huining BIAN ; Zu′an LIU ; Zhifeng HUANG ; Chuanwei SUN ; Hanhua LI ; Hongmin LUO ; Lianghua MA ; Hanxi CHEN
Chinese Journal of Burns 2017;33(4):217-223
Objective:
To explore the effects of allogeneic bone marrow mesenchymal stem cells (BMSCs) on polarization of peritoneal macrophages isolated from rats with sepsis induced by endotoxin/lipopolysaccharide (LPS).
Methods:
(1) BMSCs were isolated, cultured and purified from 5 SD rats with whole bone marrow adherent method. The third passage of cells were collected for morphologic observation, detection of expressions of stem cell surface markers CD29, CD44, CD45, and CD90 with flow cytometer, and identification of osteogenic and adipogenic differentiation. (2) Another 45 SD rats were divided into sham injury group (SI,
9.Protective effects of valproic acid on gut barrier function after major burn injury and its mechanism
Hongmin LUO ; Sen HU ; Huining BIAN ; Shaoyi ZHENG ; Bing XIONG ; Zhifeng HUANG ; Zu'an LIU ; Chuanwei SUN ; Lianghua MA ; Hanhua LI ; Wen YU ; Minghua DU ; Huada CHEN ; Wen LAI
Chinese Critical Care Medicine 2017;29(3):221-227
Objective To investigate the potential protective effects of valproic acid (VPA) on gut barrier function after major burn injury in rats and its mechanism.Methods Forty male Sprague-Dawley (SD) rats were divided into sham + normal saline (NS),sham + VPA,scald + NS,and scald + VPA groups,with 10 rats in each group.Rat with 55% total body surface area (TBSA) third-degree severe-bums model was reproduced by immersing into 80 ℃ water,and the rats in sham groups were given sham-bums by immersing into 37 ℃ water.The rats after severebums were immediately treated with 0.25 mL of 300 mg/kg VPA or NS by subcutaneous injection.Rats were sacrificed at 2 hours and 6 hours after injury,and abdominal aortic blood and ileal tissue were harvested.The levels of vascular endothelial growth factor (VEGF) were determined by enzyme-linked immunosorbent assay (ELISA).The intestinal permeability was evaluated by fluorescein isothiocyanate-dextran (FITC-dextran) determination.The histomorphological changes in gut barrier were evaluated by Chiu grading system.Levels of acetylated lysine at the ninth position of histone 3 protein (Ac-H3K9),hypoxia-inducible factor 1α (HIF-1α),zona occludens 1 (ZO-1) and myosin light chain kinase (MLCK) were determined by immunofluorescence staining and Western Blot.Results Compared with sham + NS group,rats in scald + NS group showed intestinal mucosal damage 2 hours after bum injury,as well as increased mucosal permeability,protein expression levels of HIF-1 α,VEGF,MLCK,and lowered levels of AC-H3K9 and ZO-1.These changes were much more prominent at 6 hours after injury.VPA treatment significantly attenuated the bum-induced intestinal damage.Compared with scald + NS group,the protective effects in scald + VPA group was not evident at 2 hours after injury;however,intestinal damage was much less severe at 6 hours after injury (Chiu score:2.03 ± 0.27 vs.3.12 ± 0.15),intestinal permeability was significantly decreased [FITC-dextran (μg/L):709 ± 76 vs.1138 ± 75],histone acetylation was enhanced [Ac-H3K9 (gray value):1.55 ± 0.12 vs.0.48±0.12],ZO-1 degradation was significantly inhibited (gray value:0.69 ± 0.12 vs.0.43 ± 0.16),the protein expression levels of VEGF and MLCK were significantly down-regulated [VEGF (ng/mg):51.7±3.7 vs.71.2±4.3,MLCK (gray value):1.98±0.20 vs.2.80±0.24],while the HIF-1 α protein expression levels were significantly reduced at both 2 hours and 6 hours after injury (gray value:2.50±0.39 vs.3.88±0.42 at 2 hours,1.83±0.42 vs.4.42±0.41 at 6 hours,all P < 0.05).Conclusions Severe bum injury can induce histone deacetylation,ZO-1 degradation and intestinal barrier dysfunction.VPA can improve the levels of histone acetylation and ZO-1,and protect intestinal epithelial barrier function.These may probably be mediated through inhibiting HIF-1α and its downstream gene VEGF and MLCK.
10.Effects of different-states-squatting on surface electromyography of thigh muscles and lumbar multifidus muscle
Dandan LIU ; Lianghua CHEN ; Jingjie CHEN ; Yucong ZOU ; Junqi CHEN ; Haihong LI ; Gang LIU
The Journal of Practical Medicine 2017;33(24):4069-4073
Objective This work aimed to observe averaged electromyogram(AEMG)changes in surface electromyography(sEMG)of thigh muscles and lumbar multifidus muscles in different states of squat training. Then,to find out which is the most effective for squat training. Methods A total of 46 healthy volunteers were selected and divided into two groups,namely,with and without leaning against the wall.sEMG was used to record vastus lateralis muscle,vastus medialis muscle,biceps femoris muscle,and lumbar multifidus muscles at differ-ent knee flexion angles(30°,45°,60°,and 75°). AEMG was analyzed by repeated-measures analysis of vari-ance. Results Different knee flexion angles exerted significant effects on AEMG of all tested muscles(P <0.001).Whether leaning against the wall to squat had a significant impact on AEMG of biceps femoris muscle,but not on vastus lateralis muscle,vastus medialis muscle and lumbar multifidus muscles. Significant difference was detected in interactions of knee flexion angle,and the same to the factor of with and without leaning against the wall on examined AEMG of the thigh muscle,but not for lumbar multifidus muscles. Multiple comparisons showed,the AEMG level of the tested muscles was significantly increased when the knee flexion angles was 75°. Conclusions With increased knee flexion angle,muscle activation has enheanced for vastus lateralis muscle, vastus medialis muscle,biceps femoris muscle and lumbar multifidus muscles. Squatting without leaning against the wall exerts a more marked influence on activation of biceps femoris muscle,but no activation for vastus lateralis muscle,vastus medialis muscle and lumbar multifidus muscles.

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