1.Danggui Shaoyaosan Inhibits cGAS-STING/IRF7/STAT3 Signaling Pathway to Ameliorate Neuroinflammation in Vascular Dementia
Chuyao HUANG ; Zhenwen WEI ; Ningxiang ZHENG ; Wei ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(17):1-10
ObjectiveTo investigate the anti-inflammatory effect and mechanism of Danggui Shaoyaosan (DSS) in ameliorating vascular dementia (VaD). MethodsSeventy 8-week-old C57BL/6J male mice were randomized into 7 groups: control, model, sham, positive control (donepezil, 10 mg·kg-1), and low-, medium-, and high-dose (12, 24, 36 g·kg-1, respectively) DSS groups (n=10). After drug administration, behavioral tests and cerebral blood flow detection were carried out. Enzyme-linked immunosorbent assay was used to assess the levels of interferon (IFN)-α, tumor necrosis factor (TNF)-α, interleukin (IL)-6, and CXC motif chemokine ligand 10 (CXCL10) in the brain tissue. Hematoxylin-eosin staining was employed to observe the changes in hippocampal morphology. Transcriptomics was used to predict the potential signaling mechanisms of DSS in ameliorating neuroinflammation, and Western blot and Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) were conducted to verify the expression changes of related genes. ResultsCompared with the control group and the sham group, the model group showed deceases in recognition index in the new object recognition test, movement distance and time in the target quadrant, and number of crossings in the Morris water maze test (P<0.01). In addition, the model group showed slow cerebral blood flow and down-regulated protein levels of postsynaptic density protein 95 (PSD95) and occludin (P<0.05, P<0.01), and there was no significant difference between the control group and the sham group. Compared with the model group, the DSS at each dose increased the new object recognition index, the distance and time in the target quadrant, the number of crossings, and cerebral blood flow (P<0.05, P<0.01). In terms of brain tissue injury-related protein expression and inflammatory factor content, the medium-dose DSS group had the best effect, with higher protein levels of PSD95 and occludin (P<0.05) and lower levels of IFN-α, TNF-α, and IL-6 (P<0.01) than the model group. The hippocampus of model mice showed pathological manifestations such as cell loss and disarrangement, which were alleviated after administration of DSS at each dose. Transcriptomic results indicated that interferon regulatory factor 7 (IRF7), signal transducer and activator of transcription 3 (STAT3), and bone marrow stromal cell antigen 2 (BST2) were differentially expressed genes (down-regulated) in control group and medium-dose DSS group compared with the model group. The KEGG pathway enrichment analysis showed that RIG-Ⅰ-like receptor signaling pathway, Toll-like receptor signaling pathway, cytosolic DNA-sensing pathway, and downstream stimulator of interferon genes (STING) were both upstream signaling pathways affecting IFN expression. Real-time PCR results indicated that the mRNA levels of cyclic good manufacturing practice(GMP)-adenosine monophosphate(AMP) synthase (cGAS), STING, IRF7, STAT3, and BST2 in the model group were higher than those in the sham group (P<0.05, P<0.01). The mRNA levels of all the genes in the medium-dose DSS group were down-regulated compared with those in the model group after administration (P<0.05, P<0.01). Western blot results indicated that the relative expression levels of cGAS, STING, p-IRF7, p-STAT3, and BST2 in the model group were higher than those in the sham group (P<0.05, P<0.01). The protein levels of cGAS, STING, p-IRF7, p-STAT3, and BST2 in the medium-dose DSS group were decreased compared with those in the model group (P<0.05, P<0.01). ConclusionDSS ameliorates cognitive impairment in the VaD model mice by inhibiting the cGAS-STING/IRF7/STAT3-mediated neuroinflammation.
2.Danggui Shaoyaosan Inhibits cGAS-STING/IRF7/STAT3 Signaling Pathway to Ameliorate Neuroinflammation in Vascular Dementia
Chuyao HUANG ; Zhenwen WEI ; Ningxiang ZHENG ; Wei ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(17):1-10
ObjectiveTo investigate the anti-inflammatory effect and mechanism of Danggui Shaoyaosan (DSS) in ameliorating vascular dementia (VaD). MethodsSeventy 8-week-old C57BL/6J male mice were randomized into 7 groups: control, model, sham, positive control (donepezil, 10 mg·kg-1), and low-, medium-, and high-dose (12, 24, 36 g·kg-1, respectively) DSS groups (n=10). After drug administration, behavioral tests and cerebral blood flow detection were carried out. Enzyme-linked immunosorbent assay was used to assess the levels of interferon (IFN)-α, tumor necrosis factor (TNF)-α, interleukin (IL)-6, and CXC motif chemokine ligand 10 (CXCL10) in the brain tissue. Hematoxylin-eosin staining was employed to observe the changes in hippocampal morphology. Transcriptomics was used to predict the potential signaling mechanisms of DSS in ameliorating neuroinflammation, and Western blot and Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) were conducted to verify the expression changes of related genes. ResultsCompared with the control group and the sham group, the model group showed deceases in recognition index in the new object recognition test, movement distance and time in the target quadrant, and number of crossings in the Morris water maze test (P<0.01). In addition, the model group showed slow cerebral blood flow and down-regulated protein levels of postsynaptic density protein 95 (PSD95) and occludin (P<0.05, P<0.01), and there was no significant difference between the control group and the sham group. Compared with the model group, the DSS at each dose increased the new object recognition index, the distance and time in the target quadrant, the number of crossings, and cerebral blood flow (P<0.05, P<0.01). In terms of brain tissue injury-related protein expression and inflammatory factor content, the medium-dose DSS group had the best effect, with higher protein levels of PSD95 and occludin (P<0.05) and lower levels of IFN-α, TNF-α, and IL-6 (P<0.01) than the model group. The hippocampus of model mice showed pathological manifestations such as cell loss and disarrangement, which were alleviated after administration of DSS at each dose. Transcriptomic results indicated that interferon regulatory factor 7 (IRF7), signal transducer and activator of transcription 3 (STAT3), and bone marrow stromal cell antigen 2 (BST2) were differentially expressed genes (down-regulated) in control group and medium-dose DSS group compared with the model group. The KEGG pathway enrichment analysis showed that RIG-Ⅰ-like receptor signaling pathway, Toll-like receptor signaling pathway, cytosolic DNA-sensing pathway, and downstream stimulator of interferon genes (STING) were both upstream signaling pathways affecting IFN expression. Real-time PCR results indicated that the mRNA levels of cyclic good manufacturing practice(GMP)-adenosine monophosphate(AMP) synthase (cGAS), STING, IRF7, STAT3, and BST2 in the model group were higher than those in the sham group (P<0.05, P<0.01). The mRNA levels of all the genes in the medium-dose DSS group were down-regulated compared with those in the model group after administration (P<0.05, P<0.01). Western blot results indicated that the relative expression levels of cGAS, STING, p-IRF7, p-STAT3, and BST2 in the model group were higher than those in the sham group (P<0.05, P<0.01). The protein levels of cGAS, STING, p-IRF7, p-STAT3, and BST2 in the medium-dose DSS group were decreased compared with those in the model group (P<0.05, P<0.01). ConclusionDSS ameliorates cognitive impairment in the VaD model mice by inhibiting the cGAS-STING/IRF7/STAT3-mediated neuroinflammation.
3.Simultaneous Determination of Five Nucleoside Components in Periplaneta Americana by Quantitative Analysis of Multi-components by Single Marker Method
Muyuan CHEN ; Siqi FENG ; Zhenwen QIU ; Qian LUO ; Chunzhu XIE ; Shiyu QI ; Yuechun HUANG ; Huibiao LI
Traditional Chinese Drug Research & Clinical Pharmacology 2024;35(7):1055-1060
Objective A HPLC-quantitative analysis of multi-components by single marker(QAMS)was established to determine 5 ingredients including uracil,uridine,hypoxanthine,inosine and guanosine in Periplaneta americana.Methods Separation took place on a Agilent ZORBAX SB-Aq column(250 mm×4.6 mm,5 μm)by gradient elution of methanol-0.01 mol·L-1 potassium dihydrogen phosphate at 20℃with a flow rate of 1.0 mL·min-1.The detection wavelength was 260 nm and the injection amount was 10 μL.The relative correction factors(fa/b)was calculated for the other four components with uridine as an internal standard.The content of 5 ingredients in 10 batches of Periplaneta americana was determined by QAMS.Results were compared with those of external standard method(ESM).Results Five nucleosides showed good linear relationships in their own ranges(r>0.999 5),and the average recoveries ranged from 97.0%to 100.8%.The relative correction factors of uracil,hypoxanthine,inosine and guanosine were 0.908 0,1.005 3,1.969 5 and 1.303 4,respectively.Conclusion The established method is accurate and stable.It can provide theoretical reference for the quality control of Periplaneta americana.
4.Practice of a hemodialysis alliance in the context of closed-loop hospital management
Jing QIAN ; Mengjing WANG ; Chuhan LU ; Ping CHENG ; Li NI ; Wei LIU ; Bihong HUANG ; Zhibin YE ; Zhenwen YAN ; Qianqiu CHENG ; Chen YU ; Aili WANG ; Ai PENG ; Wei XU ; Chunlai LU ; Dandan CHEN ; Xiuzhi YU ; Liyan FEI ; Jun MA ; Jialan SHEN ; Junhui LI ; Ying LI ; Lingyun CHEN ; Weifeng WU ; Rongqiang YU ; Lihua XU ; Jing CHEN
Chinese Journal of Hospital Administration 2022;38(8):595-599
Closed-loop hospital management can effectivly cope with the COVID-19 pandemic. In order to ensure the continuity of treatments for hemodialysis patients under closed-loop management and minimize possible medical and infection risks, Huashan Hospital affiliated to Fudan University and 9 hospitals in Shanghai established a hemodialysis alliance in January 2021.The alliance optimized hemodialysis resources within the region through overall planning by preparing sites, materials and personnel shifts in advance, and establishing management systems and work processes to ensure that patients could be quickly and orderly diverted to other blood dialysis centers for uninterrupted high-quality hemodialysis services, in case that some hemodialysis centers in the alliance under closed-loop management.From November 2021 to April 2022, 317 of 1 459 hemodialysis patients in the alliance were diverted to other centers for treatment, accumulating 1 215 times/cases of treatments without obvious adverse reactions. The practice could provide a reference for medical institutions to quickly establish mutual support mode under major public health events.
5.Mispronounced consonants and short-term memory among children with developmental speech sound disorder
Zhenping LIN ; Zhuoming CHEN ; Dan LI ; Yuchang LI ; Zhenwen LIANG ; Yu′en HUANG ; Wenji LUO
Chinese Journal of Physical Medicine and Rehabilitation 2020;42(5):429-433
Objective:To observe the pronunciation of consonants among children with developmental speech sound disorder and explore the correlation between mispronounced consonants and short-term memory so as to determine the pathogenesis of the disorder.Methods:Thirty-six children with developmental speech sound disorder and aged 4 to 13 years were evaluated. Their pronunciation of consonants at the phoneme and lexical levels was tested to record the error types and error rate. Twelve of the children were then randomly chosen to form a voice disorder group. Another 10 healthy counterparts constituted a control group. The short-term memory of both groups was assessed and any correlation between pronunciation and short-term memory was analyzed.Results:The children with a developmental speech sound disorder differed significantly from the controls in terms of the numbers of errors in articulating blade-alveolar, blade-palatal and velar consonants. On the phoneme level, the highest substitution error rate occurred when pronouncing lingua-palatal consonants (42.86%), followed by supradental consonants (32%). The highest distortion and non-acquisition error rates were with blade-palatal consonants (14%) and lingua-palatal consonants (9.5%). On the vocabulary level, the highest substitution, distortion, ellipsis and non-acquisition error rates appeared when pronouncing lingua-palatal and velar consonants, velar and blade-palatal consonants, supradental consonants as well as blade-palatal consonants. Significant differences were found between the phoneme and lexical levels in the substitution of supradental and blade-palatal consonants as well as in the ellipsis of blade-alveolar consonants. They were moderately associated with pronunciation level. There was, however, no significant difference in working memory span between the two groups, and no significant correlation was observed between working memory span and pronunciation level.Conclusion:The mispronunciation of consonants by children with developmental speech disorders is higher at the lexical than at the phoneme level. They mainly substitute lingua-palatal and velar consonants and elide supradental consonants, which may be related to short-term memory span.
6.Effects of Metformin on the Malignant Phenotype of Pancreatic Cancer BxPC- 3 Cells
Zhiquan HUANG ; Zhenwen WANG ; Liang ZHU
China Pharmacy 2020;31(2):202-207
OBJECTIVE:To investigate the effects of metformin on malignant phenotype of pancreatic cancer BxPC- 3 cells. METHODS:Using human pancreatic cancer BxPC- 3 cells with natural deletion of Smad4 gene as reaserch objects ,CCK-8 assay and flow cytometry were used to detect the proliferation and apoptosis of BxPC- 3 cells after treated with different doses of metformin(5,10,20 mmol/L)for 24 h. The cell survival rate and apoptosis rate were calculated. Transwell assay was used to test the migration of cells after treated with different doses of metformin (10,20 mmol/L)for 24 h. The number of migrating cells was recorded. qRT-PCR and Western blotting assay were performed to determine mRNA and protein expression of E-cadherin ,Vimentin and RGC- 32 in cells. RESULTS :Compared with control group and 5 mmol/L metformin group ,survival rate of cells were decreased significantly in 10,20 mmol/L metformin groups ,while apoptosis rate was increased significantly ;the apoptosis rate in 20 mmol/L metformin group was significantly higher than 10 mmol/L metformin group (P<0.05). Compared with control group , the number of migrating cells was decreased significantly in 10,20 mmol/L metformin groups ,and the 20 mmol/L metformin group was significantly lower than 10 mmol/L metformin group (P<0.05). Relative mRNA and protein expression of E-cadherin were increased significantly in 10,20 mmol/L metformin groups ,and relative mRNA expression of E-cadherin in 20 mmol/L metformin group was significantly higher than 10 mmol/L metformin group. Relative mRNA expression of Vimentin in 10 mmol/L metformin group ,relative mRNA and protein expression of Vimentin in 20 mmol/L metformin group ,relative mRNA and protein expression of RGC- 32 in 10,20 mmol/L metformin groups were decreased significantly ;relative mRNA and protein expression of Vimentin as well as mRNA expression of RGC- 32 in 20 mmol/L metformin group were significantly lower than 10 mmol/L metformin group (P<0.05 or P<0.01). CONCLUSIONS :Metformin can inhibit the proliferation and migration of pancreatic cancer cells through smael-independent pathways in a dose- dependent manner ,and promote their apoptosis ,which is associated with the inhibition epithelial- mesenchymal transition and the expression of RGC- 32 of pancreatic cancer.
7.CT guided percutaneous transhepatic microwave ablation for primary liver cancer in segment 9
Zaiguo WANG ; Weibiao ZHANG ; Zhenwei YE ; Yan HUANG ; Zhenwen HOU ; Yanxia MO ; Jingzhu JIANG ; Dehui HUANG ; Xiaohong HUANG ; Zhiqiang LIN ; Ailing ZHANG
Chinese Journal of Hepatobiliary Surgery 2020;26(11):825-828
Objective:To study the efficacy and safety of CT guided percutaneous transhepatic microwave ablation (PTPMWA) for primary liver cancer (PLC) in liver segment 9.Methods:A retrospective study was conducted on PLC patients between October 2013 and March 2019 at Dongguan People’s Hospital, Southern Medical University. Of 41 patients who entered into the study, there were 36 males and 5 females, with an average age of 59.1 years. These patients were diagnosed to have PLC in segment 9. The surgical related data and follow-up results were collected and analyzed.Results:All patients enrolled in the study completed the treatment procedure. CT scan was performed immediately after ablation which showed that the tumor areas to be completely covered by ablation. The duration of operation ranged from 45 to 260 (mean 91) min. The amount of bleeding during treatment was 1.0 to 5.0 (mean 1.4) ml. The complete response rate was 97.6% (40 patients) and the partial response rate was 2.4% (1 patient). The cumulative survival rates at 1, 2, 3, 4 and 5 years were 95.1%, 85.4%, 75.3%, 45.2% and 45.2%, respectively. Only 4 patients (9.8%) developed recurrence after treatment. The timings of recurrence were 1, 6, 13 and 67 months after treatment, respectively. The recurrent lesions were ablated again and complete response was obtained in all patients. There were no serious problems related to complications from ablation. The rate of postoperative complication was 7.3% (3 patients).Conclusion:PTPMWA is a novel treatment for patients with PLC in liver segment 9, the advantages of this treatment include good safety, high efficacy, low complications and local recurrence. The treatment is worthy of further future studies.
8.Clinical distribution and antibiotic resistance of Streptococcus pneumonia isolated from pediatric patients in Guangzhou
Lianfen HUANG ; Ruili XIE ; Lilan PENG ; Huamin ZHONG ; Yongqiang XIE ; Xiaoshan GUAN ; Xiaocong LI ; Xiaoguang CHEN ; Hua LI ; Zhenwen ZHOU
International Journal of Laboratory Medicine 2018;39(9):1044-1047
Objective To explore the sample type and drug resistance characteristics of Streptococcus pneu-monia(Spn)isolated from pediatric patients in Guangzhou district,and their age distribution to offer instruc-tions for prevention and clinical treatment.Methods Spn isolates were cultured and identified according to the national standard procedure for clinical laboratory operation,followed by analysis of sample type and age dis-tribution of pediatric patients with positive isolates of Spn in Guangzhou Women and Children′s Medical Cen-ter from 2013 Jan 1st to 2015 Dec 31st,drug resistance status was determined by MIC test.Results Totally, 1 243 strains of Spn were isolated,which were mainly from pediatric patients under 1 year old(42.80%).Spn isolates were mainly isolated from respiratory tract(72.81%),ear secretions(15.37%),blood(5.63%),cere-brospinal fluid(3.06%)and hydrothorax(2.01%).For all Spn isolates,the resistance rate to erythromycin, tetracycline and sulfamethoxazole was especially high as 94.93%,85.76%,73.53% respectively,with relative high resistance to penicillin G(24.70%),amoxicillin(39.59%),ceftriaxone(24.05%),meropenem(22.85%) and cefotaxime(19.89%),low resistance to quinolone antibiotics(<10.00%),and no resistance to vancomycin and linezolid.Conclusion The major age group of children with Spn infection is infants under one year old in Guangzhou,clinicians should be serious about the high resistant rate of Spn to erythromycin,tetracycline and sulfamethoxazole,the significantly increased resistant rate to penicillin,amoxicillin and ceftriaxone.Clinicians should choose antibiotics rationally according to the characteristics of drug sensitivity for better treatment.
9.Penicillin-binding proteins genotyping of penicillin resistance Streptococcus pneumonia isolated from children in Guangzhou area
Yanmei HUANG ; Xiaomin LIN ; Jialiang MAI ; Bingshao LIANG ; Yongqiang XIE ; Huamin ZHONG ; Qiulian DENG ; Zhenwen ZHOU
International Journal of Laboratory Medicine 2017;38(7):873-875,879
Objective To understand the molecular epidemiology of penicillin resistance Streptococcus pneumonia (PNSP) isolated from children in Guangzhou area to provide the experimental basis for clinical prevention and control of Streptococcus pneumonia infectious diseases.Methods Specific primers were designed according to Genebank,penicillin binding protein(PBP) genes PBP1A,PBP1B,PBP2A,PBP2B,PBP2X,PBP3 were amplified by PCR.The sequencing analysis was performed.The PCR products were digested by Hinf I,and the restriction fragment length polymorphism (RFLP) was analyzed.Results DNA of PNSP was successfully extracted,the PCR results showed that in 50 strains of PNSP,the positive rates of bacterial strains containing PBP1A,PBP1B,PBP2A,PBP2B,PBP2X and PBP3 were 48.9%,64.4%,71.1%,31.1%,40.0% and 31.1% respectively.The sequencing showed that their homologies with known sequences in GenBank were 99%,98%,100%,97%,95% and 100% respectively.Using RFLP in Hinf I showed that PBP1A,PBP1B,PBP2A and PBP3 only had one kind of genotype,PBP2B and PBP2X had two kinds of genotypes,the positive rates were 71.4%,28.6%,66.7% and 33.3% respectively.Conclusion The gene distribution of PNSP strains among children in Guangzhou is dominated by PBP2A,PBP1B and PBP1A,there are two subtypes in PBP2B,PBP2X when digested by Hinf I,in which the predominant subtype >65%.
10.Efficacy of vitamin K1 in the adjuvant treatment of pertussis syndrome
Guohong WU ; Zhenwen GAO ; Jie MA ; Dibin WANG ; Xiuming HUANG
Chinese Journal of Primary Medicine and Pharmacy 2016;23(17):2706-2708
Objective To investigate the effects of vitamin K1 in the adjuvant treatment of spasmodic cough in pertussis syndrome children,and its impacts on length of hospital stay.Methods 87 pertussis syndrome children were randomly divided into two groups,among which 41 cases in the control group were treated with the therapies such as anti -infection,respiratory support,antispasmodic,expectorant et al,while 46 cases in the treatment group were treated additionally with vitamin K1 on the basis of the control group.Then compared the duration and hospital stay of the two groups with spasmodic cough.Results The disappeared time of spasmodic cough in the treatment group was (7.22 ±1.33)d and hospital stay was (9.52 ±1.84)d,which in the control group were (15.51 ±2.73)d,(18.71 ± 3.30)d respectively,and there were statistically significant differences(t =-18.310,-16.269,all P <0.01).In the treatment group,the total effective rate was 93.5%,there were 18 cases with markedly effective,25 cases with effective and 3 cases with ineffective.Correspondingly,there were 7 cases markedly effective,23 cases effective, 11 cases ineffective,and total efficiency rate was only 73.1% in the control group,the difference was statistically significant(χ2 =4.285,P <0.05 ).Conclusion Vitamin K1 to alleviate spasmodic cough in children has a significant effect,which can reduce the duration of the cough time and hospital stay in pertussis syndrome children, and also can improve the clinical effect.

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