1.Exploring Mechanism of Chaihu Jia Longgu Mulitang in Depressive-like Rats via AMPK/SIRT1/NF-κB/NLRP3 Signaling Pathway
Guang WANG ; Xinhua SONG ; Jie YANG ; Jinyao XU ; Junhua MEI ; Chao CHEN ; Guohua CHEN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(11):144-152
ObjectiveTo investigate the effects of Chaihu Jia Longgu Mulitang(CJLM) on depression-like behaviors and neuroinflammation in rats subjected to social isolation combined with chronic unpredictable mild stress(CUMS), and to explore the potential underlying mechanisms. MethodsSixty male SD rats were randomly divided into a normal group, a model group, and low-, medium-, and high-dose CJLM groups(2.89, 5.78, 11.56 g·kg-1), as well as a fluoxetine group(10 mg·kg-1). Except for the normal group, all other groups were subjected to social isolation combined with CUMS for 63 d. During the first 35 d, depression models were established only, and from day 36 onward, modeling and drug administration were conducted simultaneously for a total intervention period of 28 d. Depression-like behaviors were evaluated using the sucrose preference test, open-field test, and forced swimming test. Hematoxylin-eosin(HE) staining was performed to observe hippocampal histomorphology. Immunohistochemistry(IHC) was used to detect the expression levels of ionized calcium-binding adapter molecule 1(Iba1) and gasdermin D(GSDMD) proteins in the hippocampus. Western blot analysis was employed to determine the protein expression levels of adenosine 5′-monophosphate(AMP)-activated protein kinase(AMPK) and phosphorylated(p)-AMPK, silent information regulator 1(SIRT1), nuclear factor-κB(NF-κB) and p-NF-κB, NOD-like receptor protein 3(NLRP3), and Caspase-1 in the hippocampus. Real-time quantitative polymerase chain reaction(Real-time PCR) was used to detect the mRNA expression levels of tumor necrosis factor-α(TNF-α), interleukin(IL)-6, and IL-1β in the hippocampus. ResultsCompared with the normal group, the model group showed a decreased sucrose preference rate(P<0.01), reduced total movement distance(P<0.01), prolonged immobility time(P<0.01), and decreased central zone residence time(P<0.01) in the open-field test, and increased immobility time in the forced swimming test(P<0.01). Hippocampal neuronal structure was damaged. The contents of Iba1 and GSDMD in the hippocampus were significantly increased(P<0.01). The protein expression levels of p-AMPK and SIRT1 in the hippocampus were significantly decreased(P<0.01), whereas the protein expression levels of p-NF-κB, NLRP3, and Caspase-1 were significantly increased(P<0.01). The mRNA expression levels of IL-1β, IL-6, and TNF-α in the hippocampus were significantly upregulated(P<0.01). Compared with the model group, the low-, medium-, and high-dose CJLM groups and the fluoxetine group all were able to reverse depression-like behavioral changes, as evidenced by increased sucrose preference rate, increased total movement distance with shortened immobility time in the open-field test, prolonged central zone residence time, and reduced immobility time in the forced swimming test(P<0.05, P<0.01). Meanwhile, hippocampal neuronal structural damage was alleviated. In the hippocampus, the expression levels of Iba1 and GSDMD were downregulated, the expression levels of p-AMPK and SIRT1 were upregulated, and the abnormal elevations of p-NF-κB, NLRP3, Caspase-1, as well as IL-1β, IL-6, and TNF-α mRNA were suppressed(P<0.05, P<0.01). ConclusionCJLM can ameliorate depression-like behaviors in rats subjected to social isolation combined with CUMS and attenuate hippocampal neuroinflammation and pyroptosis, suggesting that its effects may be associated with the regulation of AMPK/SIRT1/NF-κB/NLRP3 signaling pathway.
2.Analysis of Chemical Constituents and Blood-Absorbed Ingredients of Heihuang Chizhu Granules by UPLC-Q-TOF-MS/MS
Yiwei HUANG ; Lijiao HUANG ; Junhua HU ; Zhenzhong WANG ; Chenfeng ZHANG ; Zhongkun XU ; Yat-ing LI ; Juan FU ; Wei XIAO
Journal of Nanjing University of Traditional Chinese Medicine 2025;41(6):749-765
OBJECTIVE To analyze the chemical constituents of Heihuang Chizhu Granules and the blood composition of rats af-ter administration by UPLC-Q-TOF-MS/MS.METHODS A Waters ACQUITY UPLC HSS T3 column(3 mm×100 mm,1.8 μm)was eluted with acetonitrile-0.1%formic acid as mobile phase,and the data were collected in electrospray ion source positive and neg-ative ion mode and then identified with the reference retention time,precise molecular weight,secondary fragment ions,and references to relevant literature.RESULTS A total of 104 chemical components were identified from Heihuang Chizhu Granules,including 26 flavonoids,24 organic acids,14 triterpenoids,8 terpenoids,7 phenylpropanoids,11 monoglycosides,and 14 other components(phe-nols,alkaloids,etc.).On this basis,39 blood-entering components were identified in the plasma of rats administered via gavage,in-cluding 28 prototypes and 11 metabolites.CONCLUSION The chemical constituents of Heihuang Chizhu Granules and the compo-nents entering the blood of rats are analyzed and identified for the first time in this study,and the results provide a scientific basis for the basic research of Heihuang Chizhu Granules and the establishment of process quality control standards.
3.Efficacy of intravesical electrical stimulation and extracorporeal shock wave therapy for female non-obstructive detrusor underactivity: a reinforcement learning optimization approach
Zhen WANG ; Zhongle XU ; Junhua XI ; Yanbin ZHANG
Journal of Modern Urology 2025;30(12):1056-1063
Objective To evaluate the efficacy of intravesical electrical stimulation (IVES) and low-intensity extracorporeal shock wave therapy (Li-ESWT) in improving bladder emptying function in female patients with non-obstructive detrusor underactivity (NODU), and to further assess the clinical value of an individualized integrated treatment strategy guided by reinforcement learning (RL) optimization. Methods A total of 98 female patients diagnosed with NODU by urodynamic testing at the Department of Urology, the Second People's Hospital of Hefei, duirng Jun.2023 and Feb.2025 were prospectively enrolled. Patients were randomly assigned (1∶1∶1) to three groups:the IVES group (n=33), the Li-ESWT group (n=35), and the RL group (n=30). Clinical outcomes before and after the 4-week treatment were compared among the three groups, including peak detrusor pressure during urination (PdetQmax), maximum urinary flow rate (Qmax), post-void residual (PVR), bladder contractility index (BCI), patient perception of bladder condition-scale (PPBC-S), incontinence impact questionnaire-short form 7 (IIQ-7), urogenital distress inventory-short form 6 (UDI-6), total efficiency and satisfaction.A RL model was trained based on clinical data, with a model structure diagram and reward convergence curve plotted to validate the utility of the RL system in optimizing individualized treatment parameters. Results There were no statistically significant differences in baseline characteristics among the three groups (P>0.05). After 4 weeks of treatment, all groups demonstrated significant improvements in PdetQmax, Qmax, and BCI, along with significant reductions in PVR, PPBC-S, IIQ-7, and UDI-6 scores (all P<0.01). Notably, the RL group exhibited significantly greater improvements in PdetQmax, Qmax, and BCI, and more pronounced reductions in PVR, PPBC-S, IIQ-7, and UDI-6 scores than the IVES and Li-ESWT groups (all P<0.05). Specifically, the RL group showed the most substantial improvements in Qmax, PVR, and BCI than the other two groups (all P<0.01). The total effective rate in the RL group was 90.0% (27/30), which was higher than that of the IVES group (81.8%,27/33) and the Li-ESWT group (77.1%,27/35), but the differences were not statistically significant (χ
=2.63, P=0.27). The Li-ESWT group had a satisfaction rate of 51.4% (18/35), which was higher than that of the RL group (30.0%,9/30) and the IVES group (27.3%,9/33), but the differences were not statistically significant (χ
=6.76, P=0.34). No serious adverse events were observed in any group. After approximately 200 iterations, the reward value of the RL agent stabilized, and the individualized treatment parameters recommended further optimized bladder emptying efficiency. Conclusion Compared to IVES and Li-ESWT, the RL-optimized individualized comprehensive treatment strategy can significantly improve the bladder emptying function in women with NODU.
4.Dynamic changes of HBsAb and its predictive value in patients with chronic hepatitis B receiving antiviral therapy for clinical cure
Haiyan YANG ; Kunyan HAO ; Xieer LIANG ; Zhihong LIU ; Chunxiu ZHONG ; Junhua YIN ; Ya XU ; Leyuan WU ; Yuecheng YU ; Jinlin HOU ; Rong FAN
Chinese Journal of Hepatology 2025;33(6):551-559
Objective:To explore the predictive value of hepatitis B surface antibody (HBsAb) quantitative level for achieving hepatitis B surface antigen (HBsAg) seroclearance and serological conversion in patients with chronic hepatitis B (CHB) treated with nucleos(t)ide analogs (NAs) or interferon (IFN).Methods:A two-center prospective cohort study was conducted, including CHB patients from Nanfang Hospital Southern Medical University and Eastern Theater General Hospital treated with NAs and IFN. All patients were followed up once every three to six months. Basic clinical information and test results were collected at each follow-up. The presence or absence of HBsAg seroclearance and serological conversion rate was evaluated. HBsAg serological conversion was defined as HBsAg quantification continuously below the detection limit (<0.05 IU/mL) at two detection time points at least six months apart. HBsAg serological conversion was defined as HBsAb positivity (≥10 IU/L) at the same time as the first HBsAg seroclearance. The Kruskal-Wallis test was used to compare the quantitative data of multiple groups, and the Wilcoxon rank-sum test was used to compare the data between groups. The chi-square test was used for the count data, and the Fisher exact test was used when the chi-square test was not met. Univariate and multivariate Cox analysis was used to determine the predictors of the study endpoints, and stepwise regression was used for variable screening.Results:A total of 2 266 CHB cases were included, of which 86.5% (1 959/2 266) were NA antiviral-received population. The median treatment duration before baseline was 10.5 (2.5, 37.6) months, and the baseline HBsAg quantification was 3.1 (2.6, 3.5) log 10 IU/mL. A total of 68 cases (3.0%) had HBsAg seroclearance, and 44 cases (1.9%) achieved serological conversion after 85.0 (62.7, 97.3) months of prospective follow-up. The level and positivity rate of HBsAb showed a progressive increase 36 months before and significantly after HBsAg seroclearance. Cox regression analysis results showed that baseline HBsAb level was an independent predictor of HBsAg serological conversion ( HR=2.26, P=0.002) in the overall population, especially in the subgroup with HBsAg between 100 and 1 000 IU/mL, suggesting HBsAb level had important predictive value. In addition, the serological conversion development rate was significantly higher in the GOLDEN model favourable patients than in the unfavourable patients (11.5% vs. 0, P<0.001). Conclusion:The baseline HBsAb quantitative level can predict HBsAg seroclearance and serological conversion for patients with CHB receiving antiviral treatment, which is of significant value in long-term treatment monitoring.
5.A method for determining fluoride in water using a fully automated biochemical analyzer
Tingting LIU ; Kaiqian WANG ; Junhua ZHANG ; Yue HU ; Ji'an XIE ; Gang DING ; Weidong LI ; Shudong XU
Chinese Journal of Endemiology 2025;44(2):142-145
Objective:To establish a method for determination of fluoride in water using a fully automated biochemical analyzer (abbreviated as this method).Methods:Based on the parameters of the fully automatic biochemical analyzer, appropriate reagent volumes and reaction time were optimized to determine fluoride ions in water within the standard curve range (0.0 - 2.0 mg/L). The method was validated through evaluation of linear range, detection limit, precision, accuracy, and comparative analysis with the manual measurement results of fluoride reagent spectrophotometric method specified in the "Standard Examination Methods for Drinking Water - Part 5: Inorganic Nonmetallic indices" (GB/T 5750.5-2023).Results:Within the range of 0.0 - 2.0 mg/L fluoride mass concentration, the absolute values of the correlation coefficients were > 0.999 0 and the detection limit of this method was 0.082 mg/L. The relative standard deviation for the determination of fluoride level in water samples with low, medium and high fluoride mass concentrations was 4.03%, 2.16% and 1.68%, respectively. The spiked recovery rates were 98.1%, 99.5%, and 100.2%, respectively. There was no statistically significant difference between the results obtained by this method and manual measurement ( t = 1.07, P = 0.295). Conclusion:This method exhibits high sensitivity, accuracy, and efficiency in detecting fluoride, enabling rapid sample detection through instrumental automation instead of manual operation.
6.A method for determining fluoride in water using a fully automated biochemical analyzer
Tingting LIU ; Kaiqian WANG ; Junhua ZHANG ; Yue HU ; Ji'an XIE ; Gang DING ; Weidong LI ; Shudong XU
Chinese Journal of Endemiology 2025;44(2):142-145
Objective:To establish a method for determination of fluoride in water using a fully automated biochemical analyzer (abbreviated as this method).Methods:Based on the parameters of the fully automatic biochemical analyzer, appropriate reagent volumes and reaction time were optimized to determine fluoride ions in water within the standard curve range (0.0 - 2.0 mg/L). The method was validated through evaluation of linear range, detection limit, precision, accuracy, and comparative analysis with the manual measurement results of fluoride reagent spectrophotometric method specified in the "Standard Examination Methods for Drinking Water - Part 5: Inorganic Nonmetallic indices" (GB/T 5750.5-2023).Results:Within the range of 0.0 - 2.0 mg/L fluoride mass concentration, the absolute values of the correlation coefficients were > 0.999 0 and the detection limit of this method was 0.082 mg/L. The relative standard deviation for the determination of fluoride level in water samples with low, medium and high fluoride mass concentrations was 4.03%, 2.16% and 1.68%, respectively. The spiked recovery rates were 98.1%, 99.5%, and 100.2%, respectively. There was no statistically significant difference between the results obtained by this method and manual measurement ( t = 1.07, P = 0.295). Conclusion:This method exhibits high sensitivity, accuracy, and efficiency in detecting fluoride, enabling rapid sample detection through instrumental automation instead of manual operation.
7.Analysis of Chemical Constituents and Blood-Absorbed Ingredients of Heihuang Chizhu Granules by UPLC-Q-TOF-MS/MS
Yiwei HUANG ; Lijiao HUANG ; Junhua HU ; Zhenzhong WANG ; Chenfeng ZHANG ; Zhongkun XU ; Yat-ing LI ; Juan FU ; Wei XIAO
Journal of Nanjing University of Traditional Chinese Medicine 2025;41(6):749-765
OBJECTIVE To analyze the chemical constituents of Heihuang Chizhu Granules and the blood composition of rats af-ter administration by UPLC-Q-TOF-MS/MS.METHODS A Waters ACQUITY UPLC HSS T3 column(3 mm×100 mm,1.8 μm)was eluted with acetonitrile-0.1%formic acid as mobile phase,and the data were collected in electrospray ion source positive and neg-ative ion mode and then identified with the reference retention time,precise molecular weight,secondary fragment ions,and references to relevant literature.RESULTS A total of 104 chemical components were identified from Heihuang Chizhu Granules,including 26 flavonoids,24 organic acids,14 triterpenoids,8 terpenoids,7 phenylpropanoids,11 monoglycosides,and 14 other components(phe-nols,alkaloids,etc.).On this basis,39 blood-entering components were identified in the plasma of rats administered via gavage,in-cluding 28 prototypes and 11 metabolites.CONCLUSION The chemical constituents of Heihuang Chizhu Granules and the compo-nents entering the blood of rats are analyzed and identified for the first time in this study,and the results provide a scientific basis for the basic research of Heihuang Chizhu Granules and the establishment of process quality control standards.
8.Dynamic changes of HBsAb and its predictive value in patients with chronic hepatitis B receiving antiviral therapy for clinical cure
Haiyan YANG ; Kunyan HAO ; Xieer LIANG ; Zhihong LIU ; Chunxiu ZHONG ; Junhua YIN ; Ya XU ; Leyuan WU ; Yuecheng YU ; Jinlin HOU ; Rong FAN
Chinese Journal of Hepatology 2025;33(6):551-559
Objective:To explore the predictive value of hepatitis B surface antibody (HBsAb) quantitative level for achieving hepatitis B surface antigen (HBsAg) seroclearance and serological conversion in patients with chronic hepatitis B (CHB) treated with nucleos(t)ide analogs (NAs) or interferon (IFN).Methods:A two-center prospective cohort study was conducted, including CHB patients from Nanfang Hospital Southern Medical University and Eastern Theater General Hospital treated with NAs and IFN. All patients were followed up once every three to six months. Basic clinical information and test results were collected at each follow-up. The presence or absence of HBsAg seroclearance and serological conversion rate was evaluated. HBsAg serological conversion was defined as HBsAg quantification continuously below the detection limit (<0.05 IU/mL) at two detection time points at least six months apart. HBsAg serological conversion was defined as HBsAb positivity (≥10 IU/L) at the same time as the first HBsAg seroclearance. The Kruskal-Wallis test was used to compare the quantitative data of multiple groups, and the Wilcoxon rank-sum test was used to compare the data between groups. The chi-square test was used for the count data, and the Fisher exact test was used when the chi-square test was not met. Univariate and multivariate Cox analysis was used to determine the predictors of the study endpoints, and stepwise regression was used for variable screening.Results:A total of 2 266 CHB cases were included, of which 86.5% (1 959/2 266) were NA antiviral-received population. The median treatment duration before baseline was 10.5 (2.5, 37.6) months, and the baseline HBsAg quantification was 3.1 (2.6, 3.5) log 10 IU/mL. A total of 68 cases (3.0%) had HBsAg seroclearance, and 44 cases (1.9%) achieved serological conversion after 85.0 (62.7, 97.3) months of prospective follow-up. The level and positivity rate of HBsAb showed a progressive increase 36 months before and significantly after HBsAg seroclearance. Cox regression analysis results showed that baseline HBsAb level was an independent predictor of HBsAg serological conversion ( HR=2.26, P=0.002) in the overall population, especially in the subgroup with HBsAg between 100 and 1 000 IU/mL, suggesting HBsAb level had important predictive value. In addition, the serological conversion development rate was significantly higher in the GOLDEN model favourable patients than in the unfavourable patients (11.5% vs. 0, P<0.001). Conclusion:The baseline HBsAb quantitative level can predict HBsAg seroclearance and serological conversion for patients with CHB receiving antiviral treatment, which is of significant value in long-term treatment monitoring.
9.Epidemiological characteristics of pesticide poisoning in daily life in Tiantai County, Zhejiang Province from 2015 to 2023
Chenyi XU ; Junhua GE ; Xia CHEN ; Jiani MAO
Shanghai Journal of Preventive Medicine 2024;36(10):963-968
ObjectiveTo analyze the epidemiological characteristics and related factors of pesticide poisoning occurred in daily life in Tiantai County, Zhejiang Province from 2015 to 2023, and to provide a basis for the prevention and control of pesticide poisoning. MethodsThe information of pesticide poisoning cases was obtained from the occupational disease and occupational health information monitoring system of the China Disease Prevention and Control Information System, death data was obtained from the Zhejiang Province Chronic Disease Surveillance Information Management System, so that the poisoning incidence rate and the fatality rate were analyzed. ResultsA total of 439 cases of pesticide poisoning occurred in life were reported in Tiantai County from 2015 to 2023, of which mainly concentrated in the second quarter of each year, with an average annual incidence rate of 10.62/105 and fatality rate of 14.81%.The main cause of poisoning in < 15 years old group was accidental ingestion, while the main cause in ≥15 years old group was self-poisoning, and the difference was statistically significant (χ2=103.518, P<0.001).The average annual incidence of pesticide poisoning occurred in life in the urban area was significantly lower than that in the rural area, and the difference was statistically significant (χ2=30.871, P<0.001).The main types of pesticide poisoning were insecticides (65.15%), and paraquat had the highest fatality rate (47.73%). The results of binary logistic regression analysis indicated that age and types of pesticide poisoning may be the independent risk factors affecting the fatality of pesticide poisoning in daily life. ConclusionPesticide poisoning occurred frequently in Tiantai County over the years, and mainly occurred in summer, rural areas. Except for <15 years old group who were accidental ingestion,the other cases were mainly self-poisoning. The elderly and herbicide users should be the focus of key intervention targets. It is suggested to take personalized preventive and curative measures to intervene by age, seasons, regions and types to reduce the incidence and fatality rate of pesticide poisoning in daily life, and strengthen psychological health interventions to the cases of attempting to suicide.
10.Establishment of the fingerprints of soaking Euodiae fructus with water decoction of Radix glycyrrhizae by Zhangbang method and content determination of alkaloid
Junhua JIANG ; Jianlan CHEN ; Yan XU ; Xuping LIU
China Pharmacist 2024;27(2):220-226
Objective To establish HPLC fingerprints of soaking Euodiae fructus with water decoction of Radix glycyrrhizae by Zhangbang method(hereinafter referred to as"soaking Euodiae fructus"),and to determine the content of evodiamine,rutaecarpine and evocarpine,so as to provide the basis for the quality control and standard improvement of soaking Euodiae fructus.Methods The fingerprint of soaking Euodiae fructus was established based on wavelength switching technology,and the similarity evaluation was conducted.Taking the rutaecarpine as the internal reference,the relative correction factors of evodiamine and evocarpine were calculated by slope correction method,and the differences in measurement results between quantitative analysis of multi-components by single-marker(QAMS)method and external standard method were compared.Results The established fingerprint of soaking Euodiae fructus had a total of 20 common peaks,the similarity between it and the control fingerprint spectrum was 0.970 to 0.998,and 7 common peaks of them were identified.The contents of evodiamine and evocarpine determined by QAMS method were 1.754-7.542 mg/g and 1.281-2.455 mg/g in 10 batches of samples,and the results obtained by QAMS method and external standard method were similar.Conclusion The established HPLC method is simple,reliable,with good separation and repeatability,and can be used for quality evaluation of processed Euodiae fructus.

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