1.2,3,5,4′-tetrahydroxyldiphenylethylene-2-O-glucoside Attenuates Cerebral Ischemia-reperfusion Injury via PINK1/LETM1 Signaling Pathway
Hongyu ZENG ; Kaimei TAN ; Feng QIU ; Yun XIANG ; Ziyang ZHOU ; Dahua WU ; Chang LEI ; Hongqing ZHAO ; Yuhong WANG ; Xiuli ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(11):145-154
		                        		
		                        			
		                        			ObjectiveTo investigate the mechanism by which 2,3,5,4'-tetrahydroxyldiphenylethylene-2-O-glucoside (THSG) mitigates cerebral ischemia/reperfusion (CI/R) injury by regulating mitochondrial calcium overload and promoting mitophagy. MethodsSixty male SD rats were randomized into sham, model, SAS (40 mg·kg-1), and low-, medium- and high-dose (10, 20, 40 mg·kg-1, respectively) THSG groups, with 10 rats in each group. The middle cerebral artery occlusion/reperfusion (MCAO/R) model was established by the modified Longa suture method. An oxygen-glucose deprivation/reoxygenation (OGD/R) model was constructed in PC12 cells. Neurological deficits were assessed via Zea Longa scoring, and cerebral infarct volume was measured by 2,3,5-triphenyltetrazolium chloride (TTC) staining. Structural and functional changes of cortical neurons in MCAO/R rats were assessed by hematoxylin-eosin and Nissl staining. PC12 cell viability was detected by cell counting kit-8 (CCK-8) assay, and mitochondrial calcium levels were quantified by Rhod-2 AM. Immunofluorescence was used to detect co-localization of PTEN-induced kinase 1 (PINK1) and leucine zipper/EF-hand-containing transmembrane protein 1 (LETM1) in neurons. Transmission electron microscopy (TEM) was employed to observe mitochondrial morphology in neurons. Western blot was employed to analyze the expression of translocase of outer mitochondrial membrane 20 (TOMM20), autophagy-associated protein p62, microtubule-associated protein light chain 3 (LC3), cysteinyl aspartate-specific proteinase-9 (Caspase-9), B-cell lymphoma 2-associated protein X (Bax), and cytochrome C (Cyt C). ResultsCompared with the sham group, the model group exhibited increased infarct volume (P<0.01) and neurological deficit scores (P<0.01), neuronal structure was disrupted with reduced Nissl bodies. (P<0.01), mitochondrial swelling/fragmentation, decreased PINK1/LETM1 co-localization (P<0.01), upregulated protein levels of LC3Ⅱ/LC3Ⅰ, TOMM20, Caspase-9, Bax, and Cyt C (P<0.01), downregulated protein level of p62 (P<0.05), weakened PC12 viability (P<0.01), and elevated mitochondrial calcium level (P<0.01). Compared with the model group, THSG and SAS groups showed reduced infarct volumes (P<0.05,P<0.01) and neurological deficit scores (P<0.05,P<0.01), mitigated mitochondrial damage, and increased PINK1/LETM1 co-localization (P<0.01). Medium/high-dose THSG and SAS alleviated the neurological damage, increased Nissl bodies (P<0.05,P<0.01), downregulated the protein levels of p62, TOMM20, Caspase-9, Bax, and Cyt C (P<0.05,P<0.01), and elevated the LC3Ⅱ/LC3Ⅰ level (P<0.05,P<0.01). High-dose THSG enhanced PC12 cell viability (P<0.01), increased PINK1/LETM1 co-localization (P<0.01), and reduced mitochondrial calcium (P<0.01). ConclusionTHSG may exert the neuroprotective effect on CI/R injury by activating the PINK1-LETM1 signaling pathway, reducing the mitochondrial calcium overload, and promoting mitophagy. 
		                        		
		                        		
		                        		
		                        	
2.2,3,5,4′-tetrahydroxyldiphenylethylene-2-O-glucoside Attenuates Cerebral Ischemia-reperfusion Injury via PINK1/LETM1 Signaling Pathway
Hongyu ZENG ; Kaimei TAN ; Feng QIU ; Yun XIANG ; Ziyang ZHOU ; Dahua WU ; Chang LEI ; Hongqing ZHAO ; Yuhong WANG ; Xiuli ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(11):145-154
		                        		
		                        			
		                        			ObjectiveTo investigate the mechanism by which 2,3,5,4'-tetrahydroxyldiphenylethylene-2-O-glucoside (THSG) mitigates cerebral ischemia/reperfusion (CI/R) injury by regulating mitochondrial calcium overload and promoting mitophagy. MethodsSixty male SD rats were randomized into sham, model, SAS (40 mg·kg-1), and low-, medium- and high-dose (10, 20, 40 mg·kg-1, respectively) THSG groups, with 10 rats in each group. The middle cerebral artery occlusion/reperfusion (MCAO/R) model was established by the modified Longa suture method. An oxygen-glucose deprivation/reoxygenation (OGD/R) model was constructed in PC12 cells. Neurological deficits were assessed via Zea Longa scoring, and cerebral infarct volume was measured by 2,3,5-triphenyltetrazolium chloride (TTC) staining. Structural and functional changes of cortical neurons in MCAO/R rats were assessed by hematoxylin-eosin and Nissl staining. PC12 cell viability was detected by cell counting kit-8 (CCK-8) assay, and mitochondrial calcium levels were quantified by Rhod-2 AM. Immunofluorescence was used to detect co-localization of PTEN-induced kinase 1 (PINK1) and leucine zipper/EF-hand-containing transmembrane protein 1 (LETM1) in neurons. Transmission electron microscopy (TEM) was employed to observe mitochondrial morphology in neurons. Western blot was employed to analyze the expression of translocase of outer mitochondrial membrane 20 (TOMM20), autophagy-associated protein p62, microtubule-associated protein light chain 3 (LC3), cysteinyl aspartate-specific proteinase-9 (Caspase-9), B-cell lymphoma 2-associated protein X (Bax), and cytochrome C (Cyt C). ResultsCompared with the sham group, the model group exhibited increased infarct volume (P<0.01) and neurological deficit scores (P<0.01), neuronal structure was disrupted with reduced Nissl bodies. (P<0.01), mitochondrial swelling/fragmentation, decreased PINK1/LETM1 co-localization (P<0.01), upregulated protein levels of LC3Ⅱ/LC3Ⅰ, TOMM20, Caspase-9, Bax, and Cyt C (P<0.01), downregulated protein level of p62 (P<0.05), weakened PC12 viability (P<0.01), and elevated mitochondrial calcium level (P<0.01). Compared with the model group, THSG and SAS groups showed reduced infarct volumes (P<0.05,P<0.01) and neurological deficit scores (P<0.05,P<0.01), mitigated mitochondrial damage, and increased PINK1/LETM1 co-localization (P<0.01). Medium/high-dose THSG and SAS alleviated the neurological damage, increased Nissl bodies (P<0.05,P<0.01), downregulated the protein levels of p62, TOMM20, Caspase-9, Bax, and Cyt C (P<0.05,P<0.01), and elevated the LC3Ⅱ/LC3Ⅰ level (P<0.05,P<0.01). High-dose THSG enhanced PC12 cell viability (P<0.01), increased PINK1/LETM1 co-localization (P<0.01), and reduced mitochondrial calcium (P<0.01). ConclusionTHSG may exert the neuroprotective effect on CI/R injury by activating the PINK1-LETM1 signaling pathway, reducing the mitochondrial calcium overload, and promoting mitophagy. 
		                        		
		                        		
		                        		
		                        	
3.Modified Xiaoyaosan Alleviates Depression-like Behaviors by Regulating Activation of Hippocampal Microglia Cells in Rat Model of Juvenile Depression
Jiayi SHI ; Yun XIANG ; Ziyang ZHOU ; Dahua WU ; Feng QIU ; Chang LEI ; Hongyu ZENG ; Kaimei TAN ; Hongqing ZHAO ; Dong YANG ; Yuhong WANG ; Pengxiao GUO ; Xiuli ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(5):46-56
		                        		
		                        			
		                        			ObjectiveTo investigate the mechanism of Baihuan Xiaoyao Decoction (Xiaoyaosan added with Lilii Bulbus and Albiziae Cortex) in alleviating depression-like behaviors of juvenile rats by regulating the polarization of microglia. MethodSixty juvenile SD rats were randomized into normal control, model, fluoxetine, and low-, medium-, and high-dose (5.36, 10.71, 21.42 g·kg-1, respectively) Baihuan Xiaoyao decoction groups. The rat model of juvenile depression was established by chronic unpredictable mild stress (CUMS). The sucrose preference test (SPT) was carried out to examine the sucrose preference of rats. Forced swimming test (FST) was carried out to measure the immobility time of rats. The open field test (OFT) was conducted to measure the total distance, the central distance, the number of horizontal crossings, and the frequency of rearing. Morris water maze (MWM) was used to measure the escape latency and the number of crossing the platform. The immunofluorescence assay was employed to detect the expression of inducible nitric oxide synthase (iNOS, the polarization marker of M1 microglia) and CD206 (the polarization marker of M2 microglia). Real-time polymerase chain reaction was employed to determine the mRNA levels of iNOS, CD206, pro-inflammatory cytokines [tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6] and anti-inflammatory cytokines (IL-4 and IL-10) in the hippocampus. Western blotting was employed to determine the protein levels of iNOS and CD206 in the hippocampus. The levels of IL-4 and IL-6 in the hippocampus were detected by enzyme-linked immunosorbent assay. ResultCompared with the normal control group, the model rats showed a reduction in sucrose preference (P<0.05), an increase in immobility time (P<0.05), decreased motor and exploratory behaviors (P<0.05), and weakened learning and spatial memory (P<0.05). In addition, the model rats showed up-regulated mRNA and protein levels of iNOS and mRNA levels of IL-1β, IL-6, and TNF-α (P<0.05). Compared with the model group, Baihuan Xiaoyao decoction increased the sucrose preference value (P<0.05), shortened the immobility time (P<0.01), increased the motor and exploratory behaviors (P<0.05), and improved the learning and spatial memory (P<0.01). Furthermore, the decoction down-regulated the positive expression and protein level of iNOS, lowered the levels of TNF-α, IL-1β, and IL-6 (P<0.01), promoted the positive expression of CD206, and elevated the levels of IL-4 and IL-10 (P<0.01) in the hippocampus of the high dose group. Moreover, the high-dose Baihuan Xiaoyao decoction group had higher sucrose preference value (P<0.01), shorter immobility time (P<0.01), longer central distance (P<0.01), stronger learning and spatial memory (P<0.01), higher positive expression and protein level of iNOS (P<0.01), lower levels of TNF-α, IL-1β, and IL-6 (P<0.05, P<0.01), lower positive expression and mRNA level of iNOS (P<0.05), and higher levels of IL-4 and IL-10 (P<0.05, P<0.01) than the fluoxetine group. ConclusionBaihuan Xiaoyao decoction can improve the depression-like behavior of juvenile rats by inhibiting the M1 polarization and promoting the M2 polarization of microglia in the hippocampus. 
		                        		
		                        		
		                        		
		                        	
4.Effect of dexmedetomidine on renal function in patients with septic-associated acute kidney injury:A cohort study
Yinbi ZHENG ; Yiming SHAO ; Zhaoji LI ; Shiting LI ; Mingdi CHEN ; Wenchi ZENG ; Hongyu DONG
The Journal of Practical Medicine 2024;40(10):1423-1428
		                        		
		                        			
		                        			Objective To investigate the effect of dexmedetomidine on renal function in patients with septic-associated acute kidney injury(SAKI).Methods A prospective cohort study was conducted in 180 patients with sepsis admitted to ICU in the Second Affiliated Hospital of Guangdong Medical University from October 2021 to April 2023.According to the principle of randomized controlled trials,60 non-acute kidney injury(AKI)patients were divided into S-D group(n = 30,dexmedetomidine + conventional treatment)and S group(n = 30,conventional treatment),and the occurrence and disease score of AKI after treatment were compared between the two groups.A total of 120 AKI patients were divided into SA-D-RT group(n = 30,dexmedetomidine + blood purification + conven-tional treatment),SA-D group(n = 30,dexmedetomidine + conventional treatment),SA-RT group(n = 30,blood purification + conventional treatment)and SA group(n = 30,conventional treatment).Renal function,inflammatory factor level and disease change of the four groups were compared after treatment.Results After treatment,the inci-dence of AKI in S-D group was lower than that in S group,and the APACHEII score and SOFA score in S-D group were lower than those in S group on the 7th day after treatment(P<0.05).After 7 days of treatment,the level of Scr,BUN and CysC in the 4 groups was significantly lower than that on the 1st and 3rd day,and those in the SA-D-RT group were lower than those in the SA-D group,SA-RT group and SA group(P<0.05).After 7 days of treatment,the level of CRP,PCT,TNF-α,IL-6 and IL-1β in four groups was significantly decreased compared with on the 1st and 3rd day,and the level of CRP,PCT,TNF-α,IL-6 and IL-1β in SA-D-RT group was lower than that in SA-D group,SA-RT group and SA group(P<0.05).After 7 days of treatment,the APACHEII score and SOFA score of the four groups were significantly lower than on the 1st and 3rd day,and the scores of the SA-D group were lower than those of the SA-D group,the SA-RT group and the SA group(P<0.05).Conclusion Dexmedetomidine can effectively reduce the incidence of AKI,affect the expression level of renal function markers and inflammatory factors in serum of SAKI patients,and improve the condition of patients.
		                        		
		                        		
		                        		
		                        	
5.Changes in iron content, lipid peroxidation, and ferroptosis-related gene expression in vascular endothelial cells in vitro induced by vibration
Siheng WU ; Qingsong CHEN ; Yingshan ZENG ; Yun XIA ; Hongyu YANG
Journal of Environmental and Occupational Medicine 2024;41(5):560-566
		                        		
		                        			
		                        			Background Vascular endothelial injury is an important pathogenic step of vibration-induced hand arm vibration disease (HAVD), and long-term vibration exposure can lead to vascular endothelial cell dysfunction and cell damage. Cell ferroptosis may be one of the important mechanisms of vibration-induced vascular endothelial cell injury and HAVD. Objective To explore whether vibration can induce changes in ferroptosis-related indicators in vascular endothelial cells in vitro. Methods Human umbilical vein endothelial cells (HUVEC) were divided into four vibrationgroups and two control groups. The vibration groups were exposed to an vibration setting of 125 Hz, 6.5 m·s−2 frequency band and for different durations: 1 d 2 h (total 1 d, 2 h per day), 1 d 4 h (total 1 d, 4 h per day), 2 d 2 h (total 2 d, 2 h per day), and 2 d 4 h (total 2 d, 4 h per day), respectively. All control groups were treated the same as the experimental groups except no vibration exposure. When the cells were 80% confluent, the control groups and the corresponding experimental groups were harvested at the same time. The effects of subgroup treatments on iron, reduced glutathione (GSH), and malondialdehyde (MDA) in HUVEC were detected with a cell ferrous colorimetric test kit, a reduced GSH colorimetric test kit, and a trace MDA test kit, respectively. Real-time quantitative polymerase chain reaction (RT-PCR) was used to detect the mRNA expressions of ferroptosis-related genes acyl-CoA synthetase long chain family member 4 (ACSL4), tumor protein 53 (P53), recombinant human ferritin heavy chain (FTH1), and glutathione peroxidase 4 (GPX4). Western blot (WB) was used to detect the expression levels of ferroptosis-related proteins in HUVEC. Results Compared with the control groups, the vibration induced an increase in the iron content of HUVEC with a dose-response trend. Compared with the control groups, the reduced GSH content of HUVEC in the vibration group decreased with the increase of vibration time and frequency, and there was a dose-response trend. Compared with the control groups, the intracellular MDA content of HUVEC in the 1 d 2 h, 1 d 4 h, and 2 d 4 h vibration groups increased, and the MDA content in the 1 d 2 h and 1 d 4 h vibration group increased with time. The RT-PCR results showed that the mRNA expression levels of ACSL4 and P53 in the 1 d 4 h group increased compared with the 1 d 2 h group. Compared with the 2 d control group, the mRNA expression levels of ACSL4 in the 2 d 2 h vibration group and the 2 d 4 h vibration group increased, and the mRNA expression level of P53 in the 2 d 4 h vibration group increased. Compared with the 1 d control group, the mRNA expression levels of FTH1 and GPX4 in endothelial cells in the vibration 1 d 2 h group decreased. The WB results showed that compared with the control groups, the expression level of ferroptosis-related protein ACSL4 in endothelial cells increased in the vibration 1 d 2 h group; the expression levels of P53 in the 1 d 2 h and 2 d 4 h vibration groups increased; the expression levels of GPX4 decreased in the 1 d 4 h and 2 d 2 h vibration group, and the decrease was more obvious in the 2 d 2 h vibration group than in the 1 d 2 h vibration group; the above differences were statistically significant (P<0.05). Conclusion Vibration induces an increase in iron content, a decrease in GSH, and an increase in MDA in vascular endothelial cells in vitro, as well as mRNA and protein expressions of ferroptosis-related genes ACSL4, P53, FTH1, and GPX4.
		                        		
		                        		
		                        		
		                        	
6.Professor XU Xian's Experience in Treating Flat Warts Based on the Theory of "Bai Qi (白气) Micro-discharge"
Hongyu ZHANG ; Wanting LUO ; Yiping LI ; Qian KONG ; Xue ZENG
Journal of Traditional Chinese Medicine 2024;65(22):2294-2298
		                        		
		                        			
		                        			This paper summarized the experience in treating flat warts by Professor XU Xian based on the theory of Bai Qi (白气) micro-discharge. Flat warts is considered as a minor skin disease. Wind-damp attacking the exterior and contention between the muscles are the prerequisites for its occurrence, and failure to diffuse due to wei (卫) constraint, and qi discharge to striae and interstices are the core pathogenesis. For treatment, the core principle is to treat minor diseases with minor adjustment, and therefore, the method of dispelling wind and overcoming dampness is used to produce slight sweating and relieve the symptoms, and by taking advantage of the situation, the constraint is unblocked with diffusion and dispersion. Simultaneously, it is paid attention to not over-damaging blood vessels during treatment to avoid causing other symptoms. Self-made Modified Maxing Yigan Decoction (麻杏薏甘汤) is recommended as the basic formula for flat warts, together with self-made Xi You Formula (洗疣方) in external administration to restore defense qi, eliminate pathogenic qi and remove the warts. 
		                        		
		                        		
		                        		
		                        	
7.Experience of Zhu Renkang in Treating Chronic Eczema Based on Ziyin Chushi Decoction
Yiping LI ; Dong SHEN ; Wanting LUO ; Hongyu ZHANG ; Xue ZENG
Chinese Journal of Information on Traditional Chinese Medicine 2024;31(6):171-174
		                        		
		                        			
		                        			This article summarized Professor Zhu Renkang's experience in treating chronic eczema with Ziyin Chushi Decoction.Professor Zhu Renkang believes that eczema's onset is inseparable from pathogenic dampness.Pathogenic dampness runs through all stages of the disease.Stage of attack is dominated by excessive pathogen.Pathogenic wind attacks the exterior,while the muscular interstices is not dense,which makes nutritive yin leak out and stagnate as pathogenic dampness,then wind and dampness combined with each other.Chronic stage is dominated by yin deficiency.Long-term illness damages healthy qi,and pathogenic dampness conceals in the body,which makes deficiency of nutritive yin inside and dryness outside.Professor Zhu Renkang focuses on the characteristics of chronic eczema characterized by deficiency of yin and retention of dampness pathogens.He establishes Ziyin Chushi Decoction,and emphasizes that nutritive yin and pathogenic dampness have the same origin but different paths and can exist simultaneously.The combined utilization of nourishing yin and eliminating dampness can restore yin without helping dampness,remove dampness without harming yin,which have achieved satisfactory efficacy.
		                        		
		                        		
		                        		
		                        	
8.Anti-fatigue effect of Wujia Shengmai Yin in mice and its mechanism
Jianan HAN ; Zhuorui LIU ; Peiyong ZENG ; Shuang JIANG ; Hongyu LI
Journal of Jilin University(Medicine Edition) 2024;50(3):689-696
		                        		
		                        			
		                        			Objective:To discuss the anti-fatigue effect of Wujia Shengmai Yin,and to clarify its mechanism.Methods:Thirty-six male ICR mice were randomly devided into control group(equivalent volume of distilled water),Shengmai Yin group(500 mg·kg?1 of Shengmai Yin),and Wujia Shengmai Yin group(600 mg·kg?1 of Wujia Shengmai Yin).The body weights of the mice in various groups were detected every 7 d,and the mental states were observed.The rotating rod test and exhaustive swimming test were used to detect the duration on the rod and the swimming time to exhaustion of the mice in various groups,respectively;the levels of urea nitrogen(BUN)and lactate(LA),and the activities of lactate dehydrogenase(LDH)in serum,the levels of liver glycogen(LG),muscle glycogen(MG),and malondialdehyde(MDA),and activities of glutathione peroxidase(GSH-Px)and superoxide dismutase(SOD)in muscle tissue of the mice in various groups were detected by kits;the expression levels of glucose metabolism-related proteins in liver tissue of the mice in various groups were detected by Western blotting method.Results:Compared with before experiment,the body weights after experiment of the mice in various groups showed a increasing trend but the differences were not statistically significant(P>0.05).The rotating rod test results showed that compared with control group,the duration on the rod of the mice in Wujia Shengmai Yin group was significantly increased(P<0.01).The exhaustive swimming test results showed that compared with control group,the swimming time to exhaustion of the mice in Shengmai Yin group and Wujia Shengmai Yin group was significantly increased(P<0.01).Compared with control group,the levels of BUN in serum of the mice in Shengmai Yin group and Wujia Shengmai Yin group were significantly decreased(P<0.01),and the activities of LDH were significantly increased(P<0.01);the level of LA of the mice in Wujia Shengmai Yin group was significantly decreased(P<0.01).Compared with Shengmai Yin group,the levels of BUN and LA in the serum and LDH activity of the mice in Wujia Shengmai Yin group were significantly decreased(P<0.01).Compared with control group,the levels of LG in liver tissue and the levels of MG in muscle tissue of the mice in Shengmai Yin group and Wujia Shengmai Yin group were significantly increased(P<0.01);compared with Shengmai Yin group,the level of LG in liver tissue and the level of MG in muscle tissue of the mice in Wujia Shengmai Yin group were increased(P<0.01).Compared with control group,the activities of GSH-Px and SOD in muscle tissue of the mice in Shengmai Yin group and Wujia Shengmai Yin group were significantly increased,and the levels of MDA in Shengmai Yin group and Wujia Shengmai Yin group was significantly decreased(P<0.01);compared with Shengmai Yin group,the activities of GSH-Px and SOD in muscle tissue of the mice in Wujia Shengmai Yin group were significantly increased and the level of MDA was decreased(P<0.01).The Western blotting results showed that compared with control group,the expression levels of phosphorylated phosphoinositide 3-kinase(p-PI3K),phosphorylated protein kinase B(p-AKT),phosphorylated glycogen synthase kinase 3 beta(p-GSK3β),and glycogen synthase(GS)proteins in liver tissue of the mice in Shengmai Yin group and Wujia Shengmai Yin group were significantly increased(P<0.05 or P<0.01);compared with Shengmai Yin group,the expression levels of p-PI3K,p-AKT,p-GSK3β,and GS proteins in liver tissue of the mice in Wujia Shengmai Yin group were significantly increased(P<0.01).Conclusion:Wujia Shengmai Yin enhances the anti-fatigue effect by activating the phosphoinositide 3-kinase(PI3K)/protein kinase B(AKT)/glycogen synthase kinase 3 beta(GSK3β)signaling pathways,and improves the body's antioxidant capacity,and increases the glycogen synthesis.
		                        		
		                        		
		                        		
		                        	
9.Serum hepatitis B virus pregenomic RNA profiles in patients with chronic hepatitis B on long-term antiviral therapy
Jiali PAN ; Hao LUO ; Xiaxia ZHANG ; Yifan HAN ; Hongyu CHEN ; Zhan ZENG ; Xiaoyuan XU
Chinese Journal of Hepatology 2024;32(1):16-21
		                        		
		                        			
		                        			Objective:To explore the clinical changes in levels of the new clinical marker serum hepatitis B virus (HBV) pregenomic RNA (pgRNA) in patients with chronic hepatitis B (CHB) with long-term antiviral therapy.Methods:100 CHB cases who were initially treated with nucleos(t)ide analogues (NAs) at Peking University First Hospital were included. The levels of alanine aminotransferase (ALT), HBV DNA, hepatitis B e-antigen (HBeAg), and hepatitis B surface antigen (HBsAg) during the follow-up period were measured. The TaqMan-based real-time quantitative PCR method was used to detect serum HBV pgRNA levels. The independent sample t-test and Mann-Whitney U test were used to compare continuous variables between groups, while Pearson's χ2 test and Fisher's exact test were used to compare categorical variables. Results:HBV pgRNA levels decreased significantly in patients who developed virological responses at 48 weeks ( n = 54) during subsequent treatment compared to those who did not ( n = 46). The HBV pgRNA level was lower in HBeAg-positive patients than in HBeAg-negative patients ( P < 0.05 or P < 0.01). Patients with higher HBV DNA and HBeAg-positivity levels at baseline had a higher HBV pgRNA level following antiviral therapy. There was no statistically significant difference in HBV pgRNA levels in patients with different HBV pgRNA levels at baseline after antiviral therapy. There was no correlation between serum HBV pgRNA and HBsAg at baseline, but there was a correlation after long-term antiviral therapy, while there was a weak correlation between HBV pgRNA and HBsAg at the fifth and ninth years of antiviral therapy ( r = 0.262, P = 0.031; r = 0.288, P = 0.008). Conclusion:HBV pgRNA levels were higher with higher HBV activity in CHB patients with long-term antiviral therapy.
		                        		
		                        		
		                        		
		                        	
10.Effect of Different Cultivation Modes on Microbial Diversity and Secondary Metabolites of Rhizosphere Soil of Gentiana crassicaulis
Wuni JIADUO ; Ami JIASHI ; Hongyu MA ; Gengsheng YE ; Chen CHEN ; Kaijun GOU ; Yan REN ; Mujia LA ; Rui ZENG
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(10):192-201
		                        		
		                        			
		                        			ObjectiveTo investigate the effects of different cultivation modes on the yield of Gentiana crassicaulis and its microbial diversity and secondary metabolite content in the rhizosphere soil. MethodWith G. crassicaulis of different cultivation modes and its rhizosphere soil as the research objects, the composition of bacterial and fungal communities, dominant bacteria, and differential microorganisms in the rhizosphere soil were analyzed by high-throughput sequencing technology. HPLC was used to determine the content of iridoids in G. crassicaulis with different cultivation modes. ResultCompared with plastic film mulching, planting without mulch and intercropping of peony, white kidney bean, potato, and corn increased the yield of fresh products by 16.11%-17.68%, 22.48%-26.34%, 29.37%-32.19%, 34.82%-36.57%, and 35.34%-39.71%, respectively, and increased the yield of dry products by 19.75%-23.17%, 25.86%-29.32%, 30.18%-34.94%, 35.22%-39.87%, and 39.72%-43.73%. The total content of four iridoids, including gentiopicrin, loganic acid, sweroside, and swertiamarin, increased by 10.17%-37.83%, 5.93%-47.44%, 9.09%-28.84%, and 10.71%-28.57%, respectively. The diversity of bacterial and fungal communities in the rhizosphere soil increased significantly (P<0.05). The relative abundance of pathogenic bacteria such as Sordariomycetes, Leotiomycetes, Tremellomycetes, Eurotiomycetes, Fusarium, and Cladophialophora decreased, and the proportions of beneficial bacteria such as Proteobacteria, Acidobacteria, and Actinobacteriota increased and they gradually became the dominant bacteria. ConclusionDifferent cultivation modes can affect the yield of G. crassicaulis and its microbial diversity and iridoid content in the rhizosphere soil. Cultivation without mulch and intercropping patterns have certain advantages, which can provide theoretical references for the planting of G. crassicaulis. 
		                        		
		                        		
		                        		
		                        	
            
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