1.In vitro cultured calculus bovis alleviates cerebral ischemia/reperfusion injury through regulating microglial polarization and inhibiting NLRP3.
Tanlu CHU ; Wei ZHANG ; Jingwen CHEN ; Zeyue PAN ; Lingfeng WANG ; Xiaoming ZHONG ; Fengmei QIU ; Zhen HUANG
Journal of Zhejiang University. Medical sciences 2025;54(3):360-371
OBJECTIVES:
To investigate the effect of in vitro cultured calculus bovis (ICCB) on cerebral ischemia/reperfusion injury (CIRI) and its mechanism.
METHODS:
A CIRI rat model and a cell model were induced by middle cerebral artery occlusion (MCAO) in Sprague Dawley rats and oxygen glucose deprivation/reperfusion (OGD/R) in BV2 cells, respectively. The CIRI rat model was evaluated using the modified neurological severity score (mNSS), brain water content, and cerebral infarction volume after 1.5 h of ischemia followed by 72 h of reperfusion. Histopathological changes in the cortex and hippocampal CA1 region were observed with hematoxylin and eosin staining. Microglial polarization and NOD-like receptor thermal protein domain associated protein (NLRP) 3 inflammasome expression in the cortex were examined by immunofluorescence. BV2 cell viability was measured via MTT assay after treatment with ICCB and Nigericin. The expressions of NLRP3, ASC, caspase-1 proteins and inflammatory cytokines were detected with Western blotting in OGD/R treated BV2 cells (0.5 h OGD+24 h reperfusion) and in cells pretreated with Nigericin for 24 h.
RESULTS:
ICCB treatment significantly improved neurological function, reduced cerebral infarct volume and brain water content, and mitigated pathological damage in the cortical and hippocampal CA1 regions of rats subjected to CIRI (all P<0.05). ICCB promoted the transition of cortical microglia from M1 to M2 phenotypes and suppressed NLRP3 activation in microglial cells (all P<0.01). ICCB significantly down-regulated the expression of NLRP3, ASC, and caspase-1 proteins, and reduced the secretion of IL-18 and IL-1β in BV2 cells of OGD/R model (all P<0.01). In addition, Nigericin significantly reversed the salvage effect of ICCB on model cells (both P<0.01) and the modulation of inflammatory cytokines (P<0.05).
CONCLUSIONS
ICCB exerts a protective effect against CIRI by mitigating neuroinflammation, through the reduction of M1 microglial polarization, promotion of M2 conversion, and suppression of the NLRP3/ASC/caspase-1 signaling pathway.
Animals
;
Rats, Sprague-Dawley
;
Reperfusion Injury/prevention & control*
;
Microglia/metabolism*
;
Rats
;
NLR Family, Pyrin Domain-Containing 3 Protein
;
Brain Ischemia/metabolism*
;
Male
2.Buqi-Tongluo Decoction inhibits osteoclastogenesis and alleviates bone loss in ovariectomized rats by attenuating NFATc1, MAPK, NF-κB signaling.
Yongxian LI ; Jinbo YUAN ; Wei DENG ; Haishan LI ; Yuewei LIN ; Jiamin YANG ; Kai CHEN ; Heng QIU ; Ziyi WANG ; Vincent KUEK ; Dongping WANG ; Zhen ZHANG ; Bin MAI ; Yang SHAO ; Pan KANG ; Qiuli QIN ; Jinglan LI ; Huizhi GUO ; Yanhuai MA ; Danqing GUO ; Guoye MO ; Yijing FANG ; Renxiang TAN ; Chenguang ZHAN ; Teng LIU ; Guoning GU ; Kai YUAN ; Yongchao TANG ; De LIANG ; Liangliang XU ; Jiake XU ; Shuncong ZHANG
Chinese Journal of Natural Medicines (English Ed.) 2025;23(1):90-101
Osteoporosis is a prevalent skeletal condition characterized by reduced bone mass and strength, leading to increased fragility. Buqi-Tongluo (BQTL) decoction, a traditional Chinese medicine (TCM) prescription, has yet to be fully evaluated for its potential in treating bone diseases such as osteoporosis. To investigate the mechanism by which BQTL decoction inhibits osteoclast differentiation in vitro and validate these findings through in vivo experiments. We employed MTS assays to assess the potential proliferative or toxic effects of BQTL on bone marrow macrophages (BMMs) at various concentrations. TRAcP experiments were conducted to examine BQTL's impact on osteoclast differentiation. RT-PCR and Western blot analyses were utilized to evaluate the relative expression levels of osteoclast-specific genes and proteins under BQTL stimulation. Finally, in vivo experiments were performed using an osteoporosis model to further validate the in vitro findings. This study revealed that BQTL suppressed receptor activator of NF-κB ligand (RANKL)-induced osteoclastogenesis and osteoclast resorption activity in vitro in a dose-dependent manner without observable cytotoxicity. The inhibitory effects of BQTL on osteoclast formation and function were attributed to the downregulation of NFATc1 and c-fos activity, primarily through attenuation of the MAPK, NF-κB, and Calcineurin signaling pathways. BQTL's inhibitory capacity was further examined in vivo using an ovariectomized (OVX) rat model, demonstrating a strong protective effect against bone loss. BQTL may serve as an effective therapeutic TCM for the treatment of postmenopausal osteoporosis and the alleviation of bone loss induced by estrogen deficiency and related conditions.
Animals
;
NFATC Transcription Factors/genetics*
;
Drugs, Chinese Herbal/pharmacology*
;
Ovariectomy
;
Osteoclasts/metabolism*
;
Female
;
Osteogenesis/drug effects*
;
Rats, Sprague-Dawley
;
Rats
;
NF-kappa B/genetics*
;
Osteoporosis/genetics*
;
Signal Transduction/drug effects*
;
Bone Resorption/genetics*
;
Cell Differentiation/drug effects*
;
Humans
;
RANK Ligand/metabolism*
;
Mitogen-Activated Protein Kinases/genetics*
;
Transcription Factors
3.Laboratory Diagnosis and Molecular Epidemiological Characterization of the First Imported Case of Lassa Fever in China.
Yu Liang FENG ; Wei LI ; Ming Feng JIANG ; Hong Rong ZHONG ; Wei WU ; Lyu Bo TIAN ; Guo CHEN ; Zhen Hua CHEN ; Can LUO ; Rong Mei YUAN ; Xing Yu ZHOU ; Jian Dong LI ; Xiao Rong YANG ; Ming PAN
Biomedical and Environmental Sciences 2025;38(3):279-289
OBJECTIVE:
This study reports the first imported case of Lassa fever (LF) in China. Laboratory detection and molecular epidemiological analysis of the Lassa virus (LASV) from this case offer valuable insights for the prevention and control of LF.
METHODS:
Samples of cerebrospinal fluid (CSF), blood, urine, saliva, and environmental materials were collected from the patient and their close contacts for LASV nucleotide detection. Whole-genome sequencing was performed on positive samples to analyze the genetic characteristics of the virus.
RESULTS:
LASV was detected in the patient's CSF, blood, and urine, while all samples from close contacts and the environment tested negative. The virus belongs to the lineage IV strain and shares the highest homology with strains from Sierra Leone. The variability in the glycoprotein complex (GPC) among different strains ranged from 3.9% to 15.1%, higher than previously reported for the seven known lineages. Amino acid mutation analysis revealed multiple mutations within the GPC immunogenic epitopes, increasing strain diversity and potentially impacting immune response.
CONCLUSION
The case was confirmed through nucleotide detection, with no evidence of secondary transmission or viral spread. The LASV strain identified belongs to lineage IV, with broader GPC variability than previously reported. Mutations in the immune-related sites of GPC may affect immune responses, necessitating heightened vigilance regarding the virus.
Humans
;
China/epidemiology*
;
Genome, Viral
;
Lassa Fever/virology*
;
Lassa virus/classification*
;
Molecular Epidemiology
;
Phylogeny
4.Clinical efficiency of lumbar hernia repair based on path planning
Lisheng WU ; Chen PAN ; Xiaohan WEI ; Zhen REN ; Hu LIU
Chinese Journal of Digestive Surgery 2024;23(9):1214-1219
Objective:To investigate the clinical efficiency of lumbar hernia repair based on path planning.Methods:The retrospective and descriptive study was conducted. The clinical data of 35 patients with lumbar hernia who were admitted to The First Affiliated Hospital of University of Science and Technology of China from November 2016 to March 2024 were collected. There were 14 males and 21 females, aged (61±8)years. According to preoperative computerized tomography examination of the hernia defect diameter, patients with a defect diameter <4 cm underwent enhan-ced field laparoscopic total extraperitoneal repair (eTEP), patients with a defect diameter of 4-8 cm underwent laparoscopic partial extraperitoneal repair (TAPE), and patients with a defect diameter >8 cm underwent open preperitoneal mesh repair (Sublay). Measurement data with normal distribu-tion were represented as Mean± SD, and comparison of three groups were conducted using the one-way ANOVA or Kruskal Wallis test, and Bonferroni correction was used for pariwise comparison. Measurement data with skewed distribution were represented as M(range). Count data were descri-bed as absolute numbers or percentages, and comparison between groups was conducted using the chi-square test or Fisher exact probability. Results:(1) Operation conditions. Of 35 patients, there were 15 cases undergoing eTEP, of 7 males and 8 females and 12 cases of left hernia and 3 cases of right hernia, with operation time of (92±44)minutes and the duration of postoperative hospital stay of (5.6±2.8)days. There were 17 cases undergoing TAPE, of 5 males and 12 females and 9 cases of left hernia, 7 cases of right hernia and 1 case of bilateral hernia, with operation time of (114±56)minutes and the duration of postoperative hospital stay of (6.4±3.0) days. There were 3 cases undergoing Sublay, of 2 males and 1 female and 1 case of left hernia and 2 cases of right hernia, with operation time of (150±55)minutes and the duration of postoperative hospital stay of (12.3±7.8)days. There were significant differences in the duration of postoperative hospital stay among the three groups ( F=4.83, P<0.05). (2) Follow-up. All 35 patients were followed up for 40.5(range, 3.0-91.0)days. None of patient underwent postoperative complications such as serous swelling, incision infection, intestinal fistula, intestinal obstruction, or puncture hematoma, and no recurrence of lumbar hernia occurred. One patient who underwent TAPE had postoperative abdominal distension, and was cured by symptomatic treatment. Cases with acute pain within postoperative 3 months were 0, 5, 2 in patients undergoing eTEP, TAPE, Sublay, respectively, showing significant differences among them ( χ2=8.69, P<0.05). Results of pariwise comparison showed that there was a significant difference in acute pain within postoperative 3 months between patients undergoing eTEP and Sublay ( P<0.05), and there was a significant difference in acute pain within postoperative 3 months between patients undergoing eTEP and TAPE ( P<0.05); Cases with chronic pain after postoperative 3 months were 0, 1, 1 in patients undergoing eTEP, TAPE, Sublay, respectively, showing no significant difference among them ( χ2=4.00, P>0.05). Conclusion:It is safe and feasible to formulate the surgical method according to the defect diameter of lumbar hernia.
5.Application of multi-disciplinary team model in the management of hospi-tal antimicrobial management
Lin-Li PAN ; Da-Wei WU ; Shan-Shan LIU ; Jie PANG ; Xiu-Zhen ZOU
Chinese Journal of Infection Control 2024;23(6):719-724
Objective To explore the application effect of multi-disciplinary team(MDT)model in hospital antimi-crobial management.Methods Relevant data on antimicrobial use in hospitalized patients in a hospital from January 2021 to December 2022 were analyzed retrospectively,January-December 2021 adopted conventional management mode and was as the control group,January-December 2022 adopted MDT management model and was as the inter-vention group.Antimicrobial therapy relevant indicators between two groups of patients were compared.Results After adopting the MDT management model,pathogen detection rate before the therapeutic antimicrobial use in the intervention group(73.62%)was higher than that in the control group(70.56%),difference was statistically sig-nificant(P<0.001).Pathogen detection rate related to healthcare-associated infection diagnosis was 87.98%in the control group and 88.89%in the intervention group,with no statistically significant difference between two groups(P>0.05).Pathogen detection rate before combined use of key antimicrobial agents in the intervention group(93.94%)was higher than that in the control group(92.00%),difference was statistically significant(P<0.05).Antimicro-bial use rate in hospitalized patients and use rate of prophylactic antimicrobial agents in class Ⅰ incision surgery de-creased from 38.03%and 21.03%to 32.78%and 10.30%,respectively,with statistically significant differences(both P<0.05).The amount and intensity of antimicrobial use in hospitalized patients in the intervention group de-creased.The implementation rate of bundled prevention and control measures for multidrug-resistant organism(MDRO)after intervention was significantly higher than that of the control group,with statistically significant differences(all P<0.05).MDRO detection rate decreased from 34.70%to 32.37%,difference was statistically significant(P=0.027).there was no significant change in the MDRO case infection rate.Conclusion The MDT model can effectively improve the standardized management of antimicrobial agents,promote the rational use of anti-microbial agents in clinical practice,and prevent bacterial resistance.
6.Establishment and evaluation of a rapid PCR-colloidal gold test strip method for the detection of Fritillaria ussuriensis
Yu-he MA ; Cong-hui SHANG ; Qiu-he MA ; Tao LI ; Yue LIU ; Bei-zhen PAN ; Li-jun GAO ; Ming-cheng LI ; Wei XIA ; Yong-mei QU
Acta Pharmaceutica Sinica 2024;59(6):1773-1778
This study design of specific identification primers for the ITS2 sequence of
7.Experimental study on anti-fatigue effect of Polysaccharides of Panax notoginseng
Pan-Pan WEI ; Zi-Jun YAN ; Meng-Yue DENG ; Die XIA ; Yu-Zhen DING ; Lei ZHANG ; Tong CHEN
The Chinese Journal of Clinical Pharmacology 2024;40(1):87-91
Objective To explore the effect of Polysaccharides of Panax notoginseng(PPN)on anti-exercise fatigue in mice.Methods One hundred male KM mice were randomly divided into negative control group,positive control group and experimental-L,-M,-H groups,with 20 cases per group.Experimental-L,-M,-H groups was given 100,200,400 mg·kg-1 PPN,respectively;positive control group was given 200 mg·kg-1 vitamin C;negative control group was given 0.1 mL·10 g-1 0.9%NaCl.Five groups were gavaged once a day for 28 days.After the last administration,the loaded swimming time was measured;after 90 minutes of the unloaded swimming test,the mice were allowed to rest for 30 minutes,the levels of lactic acid(LD),blood urea nitrogen(BUN),glycogen,and malondialdehyde(MDA)were measured,the safety of PPN with organ indices and histopathology.Results LD levels in negative control group,positive control group and experimental-L,-M,-Hgroupswere(4.76±0.84),(2.86±0.34),(3.00±0.69),(2.35±0.65)and(1.39±0.48)mg·kg-1;BUN contents were(13.65±1.25),(12.55±0.91),(12.12±1.24),(11.06±1.30)and(9.85±1.05)mmol·L-1;liver glycogen contents were(3.24±0.56),(11.11±2.16),(5.61±1.41),(6.60±1.49)and(12.05±2.25)mg·g-1;MDA levels were(2.36±0.21),(1.23±0.41),(1.93±0.23),(1.73±0.21)and(1.04±0.18)mg prot·mL-1.Compared with negative control group,the differences of above indexes in the positive control group and experimental-L,-M,-H groups were statistically significant(P<0.05,P<0.01,P<0.001).Conclusion PPN can increase exercise endurance in mice and has an anti-fatigue effect.This study provides a theoretical basis for the application of PPN in the field of anti-fatigue research.
8.Antioxidant activity and organ protection of Panax notoginseng polysaccharide on oxidative damage and aging model mice
Meng-Yue DENG ; Pan-Pan WEI ; Ming LI ; Zi-Jun YAN ; Die XIA ; Yu-Zhen DING ; Lei ZHANG ; Tong CHEN
The Chinese Journal of Clinical Pharmacology 2024;40(6):889-893
Objective To study the antioxidant activity and organ protection of different components of Panax notoginseng polysaccharide(PNPS)in D-galactose-induced oxidative damage aging model mice.Methods KM mice were randomly divided into normal group,model group,vitamin C(VC)group(given 200 mg·kg-1 VC),crude polysaccharide from Panax notoginseng(CPPN)group,neutral polysaccharide from Panax notoginseng(NPPN)group and acidic polysaccharide from Panax notoginseng(APPN-Ⅰ,APPN-Ⅱ,APPN-Ⅲ)group(given 400 mg·kg-1 CPPN,NPPN,APPN-Ⅰ,APPN-Ⅱ,APPN-Ⅲ,respectively).Except for the normal group,oxidative injury aging mouse models were established by intraperitoneal injection of 1 g·kg-1 D-galactose.The mice were sacrificed after continuous administration for 42 days,and serum and liver homogenate were prepared.Malondialdehyde(MDA)was determined by thiobarbituric acid method;superoxide dismutase(SOD)was determined by tetrazole salt method;glutathione peroxidase(GSH-Px)was determined by double antibody sandwich method.Results Serum SOD in the normal group,model group,VC group,CPPN group,NPPN group and APPN-Ⅰ,APPN-Ⅱ,APPN-Ⅲ groups were(15.07±0.69),(12.79±1.51),(15.56±1.01),(13.69±0.96),(14.27±0.64),(14.31±0.99),(14.18±0.79)and(15.85±0.89)U·mL-1;serum GSH-Px were(105.35±4.97),(90.36±4.31),(111.51±7.00),(113.03±8.06),(118.77±5.19),(123.60±8.08),(131.65±3.60)and(149.22±13.32)ng·L-1;serum MDA were(1.72±0.26),(4.16±0.92),(2.26±0.59),(2.82±0.47),(2.46±0.50),(1.98±0.41),(2.39±0.39)and(2.07±0.24)nmol·mL-1;the liver SOD were(234.22±3.84),(205.04±7.28),(234.63±6.37),(214.99±17.66),(234.13±3.63),(234.63±3.44),(233.87±5.63)and(235.42±2.33)U·mgprot-1;liver GSH-Px were(274.27±23.72),(207.00±15.22),(257.68±16.39),(249.79±18.78),(252.62±10.92),(256.25±21.83),(261.20±17.52)and(263.16±17.98)ng·L-1;liver MDA were(35.70±3.52),(49.65±6.32),(36.15±2.48),(39.17±4.29),(37.40±6.19),(35.34±4.06)and(35.90±5.36),(33.31±7.64)nmol·mgprot-1.Compared with the normal group,SOD,GSH-Px in serum and liver of mice in the model group were significantly reduced,and the content of MDA was significantly increased(all P<0.01).After treatment with different components of Panax notoginseng polysaccharide,the oxidative indicators in mice were significantly improved,among which APPN-Ⅲ have the best antioxidant activity,which could significantly increase the activities of SOD,GSH-Px in serum and liver,and reduce the content of MDA(all P<0.01).Conclusion Different components of Panax notoginseng polysaccharide have antioxidant activity and organ protection in vivo,among which APPN-Ⅲ has the best antioxidant activity and has a good organ protection effect.
9.Study on Herbal Textual Research and Identification of Macleaya Cordata
Wei ZHANG ; Zeyue PAN ; Lei ZHANG ; Shujie DONG ; Fengmei Qiu ; Zhen HUANG
Chinese Journal of Modern Applied Pharmacy 2024;41(6):750-759
OBJECTIVE
To study on the identification of traditional Chinese medicine of Macleaya cordata(M. cordata) and its similar varieties.
METHODS
By consulting the ancient herbal books and modern literature, this paper systematically combs and studies the M. cordata. The morphological identification, microscopic identification, physiochemical identification, molecular identification were used to identify M. cordata and its similar varieties.
RESULTS
Obtained M. cordata herbal textual research data. There were some differences between M. cordata and Macleaya microcarpa(M. microcarpa) and other similar varieties in traits, microscopic, physicochemical and molecular characteristics. Molecular identification results showed that the length of the rbcL gene of M. cordata were 600 bp to 603 bp, with the average GC content ranging from 43.95% to 44.28%. There were significant differences in the variation sites between M. cordata and other similar varieties, and the variation sites with M. microcarpa were the least. The interspecific genetic distance between M. cordata and its similar varieties was greater than its maximum intraspecific genetic distance. NJ analysis results of rbcL could effectively distinguish M. cordata from other similar varieties accurately and quickly. There were significant differences in the secondary structure of rbcL between M. cordata and its similar varieties.
CONCLUSION
The traditional Chinese medicine identification methods of M. cordata, M. microcarpa and other similar varieties are constructed, which provides experimental basis for the variety identification of M. cordata and the subsequent development of traditional Chinese medicine resources.
10.Effect of Cinobufacini on HepG2 cells based on CXCL5/FOXD1/VEGF pathway
Xiao-Ke RAN ; Xu-Dong LIU ; Hua-Zhen PANG ; Wei-Qiang TAN ; Tie-Xiong WU ; Zhao-Quan PAN ; Yuan YUAN ; Xin-Feng LOU
Chinese Pharmacological Bulletin 2024;40(12):2361-2368
Aim To investigate the impact of Cinobu-facini on the proliferation,invasion,and apoptosis of HepG2 cells and the underlying mechanism.Methods The proliferation of HepG2 cells was assessed using the CCK-8 method following treatment with Cinobufaci-ni.The invasion capability of HepG2 cells was evalua-ted through Transwell assay after exposure to Cinobufa-cini.The apoptosis rates of HepG2 cells post Cinobufa-cini intervention were measured using flow cytometry,and the expression levels of VEGF in the culture medi-um of HepG2 cells were determined using enzyme-linked immunoassay.Furthermore,qRT-PCR and Western blot analyses were conducted to assess the im-pact of Cinobufacini on mRNA and protein expression levels related to the CXCL5/FOXD1/VEGF pathway.The interaction between CXCL5 and FOXD1 was inves-tigated via co-immunoprecipitation.Results Cinobufa-cini treatment led to a gradual decrease in HepG2 cell viability in a dose-dependent manner compared to the control group(P<0.05).Moreover,Cinobufacini sig-nificantly suppressed HepG2 cell invasion(P<0.05)while enhancing cell apoptosis(P<0.05).Notably,Cinobufacini exhibited inhibitory effects on the CX-CL5/FOXD1/VEGF pathway,as evidenced by re-duced expression of related mRNA and proteins(P<0.05).FOXD1 was identified as the binding site of CXCL5.Overexpression of CXCL5 resulted in in-creased proliferation and VEGF secretion by HepG2 cells(P<0.05),and increased expression of FOXD1 and VEGF(P<0.05).However,Cinobufacini inter-vention effectively inhibited liver cancer cell prolifera-tion and invasion(P<0.05),promoted apoptosis(P<0.05),reduced VEGF secretion by HepG2 cells(P<0.05),and downregulated the expression of CXCL5 and FOXD1 in HepG2 cells(P<0.05);but com-pared with the unexpressed group of Cinobufacini,its ability to inhibit cell activity was weakened(P<0.05),and its ability to inhibit the expression of CX-CL5,FOXD1,and VEGF was weakened(P<0.05).Conclusion Cinobufacini may inhibit HepG2 cell pro-liferation and invasion and promote HepG2 cell apopto-sis by regulating the CXCL5/FOXD1/VEGF pathway.


Result Analysis
Print
Save
E-mail