1.Alleviation of Ulcerative Colitis by Shaoyaotang via Inhibiting Glycolysis Through SIRT6/HIF-1α Pathway
Yiling XIA ; Hui CAO ; Dongsheng WU ; Bo ZOU ; Erle LIU ; Yiwen WANG ; Shaijin JIANG ; Yiqian YU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(11):10-19
ObjectiveTo investigate the role of silent information regulatory protein (SIRT6)/hypoxia-inducible factor-1α (HIF-1α) pathway in regulating the reprogramming of glucose metabolism in ulcerative colitis (UC) and the mechanism of intervention of Shaoyaotang. MethodsForty-eight c57bL/6 mice were randomly divided into a blank group, a model group, a Mesalazine group (0.42 g·kg-1), a Shaoyaotang group (31.08 g·kg-1), an inhibitor group (OSS-128167, 50 mg·kg-1), and an inhibitor + Shaoyaotang group (50 mg·kg-1 OSS-128167 + 31.08 g·kg-1 Shaoyaotang). A UC model was established by the administration of 2.5% dextran sulfate sodium (DSS) solution for mice in other groups for 7 d, except for the blank group. The mice in each group were treated with saline, Mesalazine, Shaoyaotang, inhibitor, and inhibitor + Shaoyaotang, respectively, for 7 d. The mice were necropsied 24 h after the last administration of the drug. The blood was collected from the orbital region, and colon tissue was taken. Hematoxylin-eosin (HE) staining was used to observe the pathological changes in colon tissue. Enzyme-linked immunosorbent assay (ELISA) was employed to detect serum interleukin (IL)-10, IL-17, and IL-6 levels. A biochemical method was used to detect glucose and lactate dehydrogenase A (LDHA) levels. Immunohistochemistry (IHC) was employed to detect IL-22 and transforming growth factor-β1 (TGF-β1) levels in colon tissue, and Western blot and real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) were used to detect relative protein and mRNA expressions of SIRT6, HIF-1α, and LDHA. ResultsCompared with those of the blank group, disease activity index (DAI) scores of mice in the model group and inhibitor group were significantly increased (P<0.01). The length of colon tissue was significantly shortened, and colon tissue was congested and eroded. The pathohistological scores were significantly increased (P<0.01). The levels of serum inflammatory factors IL-17 and IL-6 were significantly elevated, and the levels of IL-10 were significantly decreased (P<0.01). The protein expressions of IL-22 and TGF-β1 were significantly reduced in colon tissue (P<0.01). The relative protein and mRNA expressions of SIRT6 were significantly decreased (P<0.01), and the relative protein and mRNA expressions of HIF-1α and LDHA and the contents of glucose and lactate were significantly elevated (P<0.01). The level of inflammation in the colon of the mice in the inhibitor group was more severe than that in the model group (P<0.01). Compared with the model group, the Mesalazine group, the Shaoyaotang group, and the inhibitor + Shaoyaotang group showed reduced colonic injury, significant decrease in serum IL-17 and IL-6, significant increase in IL-10 (P<0.01), significant increase in the protein expressions of IL-22 and TGF-β1 in colon tissue (P<0.01), significant increase in the protein expressions of SIRT6 and the relative mRNA expressions (P<0.01), and significant reduction in the protein expressions of HIF-1α and LDHA, the relative mRNA expressions, and the contents of glucose and lactate (P<0.01). Compared with those in the Shaoyaotang group, the serum IL-17 and IL-6 were significantly increased, and IL-10 was significantly decreased in the inhibitor + Shaoyaotang group (P<0.01). The protein expressions of IL-22 and TGF-β1 in colon tissue were significantly decreased (P<0.01). The expressions of SIRT6 protein and the relative mRNA expressions were significantly decreased (P<0.01). The protein expressions of HIF-1α and LDHA, the relative mRNA expressions, and the contents of glucose and lactate were significantly elevated (P<0.01). However, the difference between the Shaoyaotang group and the Mesalazine group was not significant. ConclusionShaoyaotang can effectively treat DSS-induced mice with UC through the SIRT6/HIF-1α pathway, and its mechanism of action may be related to the regulation of the SIRT6/HIF-1α pathway and glucose metabolism reprogramming and the inhibition of glycolysis.
2.Alleviation of Ulcerative Colitis by Shaoyaotang via Inhibiting Glycolysis Through SIRT6/HIF-1α Pathway
Yiling XIA ; Hui CAO ; Dongsheng WU ; Bo ZOU ; Erle LIU ; Yiwen WANG ; Shaijin JIANG ; Yiqian YU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(11):10-19
ObjectiveTo investigate the role of silent information regulatory protein (SIRT6)/hypoxia-inducible factor-1α (HIF-1α) pathway in regulating the reprogramming of glucose metabolism in ulcerative colitis (UC) and the mechanism of intervention of Shaoyaotang. MethodsForty-eight c57bL/6 mice were randomly divided into a blank group, a model group, a Mesalazine group (0.42 g·kg-1), a Shaoyaotang group (31.08 g·kg-1), an inhibitor group (OSS-128167, 50 mg·kg-1), and an inhibitor + Shaoyaotang group (50 mg·kg-1 OSS-128167 + 31.08 g·kg-1 Shaoyaotang). A UC model was established by the administration of 2.5% dextran sulfate sodium (DSS) solution for mice in other groups for 7 d, except for the blank group. The mice in each group were treated with saline, Mesalazine, Shaoyaotang, inhibitor, and inhibitor + Shaoyaotang, respectively, for 7 d. The mice were necropsied 24 h after the last administration of the drug. The blood was collected from the orbital region, and colon tissue was taken. Hematoxylin-eosin (HE) staining was used to observe the pathological changes in colon tissue. Enzyme-linked immunosorbent assay (ELISA) was employed to detect serum interleukin (IL)-10, IL-17, and IL-6 levels. A biochemical method was used to detect glucose and lactate dehydrogenase A (LDHA) levels. Immunohistochemistry (IHC) was employed to detect IL-22 and transforming growth factor-β1 (TGF-β1) levels in colon tissue, and Western blot and real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) were used to detect relative protein and mRNA expressions of SIRT6, HIF-1α, and LDHA. ResultsCompared with those of the blank group, disease activity index (DAI) scores of mice in the model group and inhibitor group were significantly increased (P<0.01). The length of colon tissue was significantly shortened, and colon tissue was congested and eroded. The pathohistological scores were significantly increased (P<0.01). The levels of serum inflammatory factors IL-17 and IL-6 were significantly elevated, and the levels of IL-10 were significantly decreased (P<0.01). The protein expressions of IL-22 and TGF-β1 were significantly reduced in colon tissue (P<0.01). The relative protein and mRNA expressions of SIRT6 were significantly decreased (P<0.01), and the relative protein and mRNA expressions of HIF-1α and LDHA and the contents of glucose and lactate were significantly elevated (P<0.01). The level of inflammation in the colon of the mice in the inhibitor group was more severe than that in the model group (P<0.01). Compared with the model group, the Mesalazine group, the Shaoyaotang group, and the inhibitor + Shaoyaotang group showed reduced colonic injury, significant decrease in serum IL-17 and IL-6, significant increase in IL-10 (P<0.01), significant increase in the protein expressions of IL-22 and TGF-β1 in colon tissue (P<0.01), significant increase in the protein expressions of SIRT6 and the relative mRNA expressions (P<0.01), and significant reduction in the protein expressions of HIF-1α and LDHA, the relative mRNA expressions, and the contents of glucose and lactate (P<0.01). Compared with those in the Shaoyaotang group, the serum IL-17 and IL-6 were significantly increased, and IL-10 was significantly decreased in the inhibitor + Shaoyaotang group (P<0.01). The protein expressions of IL-22 and TGF-β1 in colon tissue were significantly decreased (P<0.01). The expressions of SIRT6 protein and the relative mRNA expressions were significantly decreased (P<0.01). The protein expressions of HIF-1α and LDHA, the relative mRNA expressions, and the contents of glucose and lactate were significantly elevated (P<0.01). However, the difference between the Shaoyaotang group and the Mesalazine group was not significant. ConclusionShaoyaotang can effectively treat DSS-induced mice with UC through the SIRT6/HIF-1α pathway, and its mechanism of action may be related to the regulation of the SIRT6/HIF-1α pathway and glucose metabolism reprogramming and the inhibition of glycolysis.
3.Establishment and evaluation of a method for detection of ASFV antigen by doub-le-antibody sandwich ELISA
Qixuan LI ; Huixian YUE ; Yiqian JIANG ; Yanyan ZHANG ; Teng CHEN ; Shuchao WANG ; Shoufeng ZHANG ; Rongliang HU
Chinese Journal of Veterinary Science 2024;44(8):1579-1584,1592
African swine fever(ASF)is an acute and highly pathogenic hemorrhagic disease of pigs,causing huge economic losses to pig industry.In order to quantitatively detect clinical samples of ASF and inactivated ASFV antigens,the IgG of ASF positive serum was used as capture anti-body and the HRP-labeled p72 monoclonal antibody was used as detecting antibody.The standard curve was drawn with the cell-cultured ASFV,and a sandwich ELISA detection of antigen was es-tablished.The specificity,sensitivity and stability of the method were evaluated.The effects of dif-ferent inactivation methods and adjuvant addition on antigen detection were further evaluated.The results showed that the minimum detection limits of the recombinant protein and the ASFV were 0.1 mg/L and 103.7 TCID50/mL,respectively.There was no cross-reaction with five common porcine pathogenic viruses,and the coefficient variations between batches was less than 10%.The total co-incidence rate with real-time fluorescence quantitative PCR was 92%(23/25).The sensitivity of antigen detection was significantly reduced when antigen was treated by BEI inactivation,and the detection results were severely interfered by aluminum adjuvant and nano-adjuvant.In summary,the sandwich ELISA antigen detection method established is specific,sensitive,and repeatable,with a good consistency to the qPCR method,which provides an effective clinical diagnostic meth-od for ASFV antigen.
4.Research progress of PLGA microspheres in stomatology
Yuqian LI ; Jing JIANG ; Menghan CHEN ; Yiqian SHAO ; Jianying FENG
STOMATOLOGY 2024;44(11):876-880
Compared with other drug loading methods,PLGA drug-loaded microspheres have the advantages of sustained and con-trolled release ability,biodegradability,drug release stability,and targeted delivery function,which have become a research hotspot in pharmaceutics.With the improvement of the preparation technology and the quality of microspheres,they have been gradually promoted in clinical practice,and have achieved remarkable results in the treatment of tumors,osteoarthritis,diabetes,eye diseases and so on.However,there is still a lack of systematic review on the application of PLGA microspheres in stomatology.This article aims to intro-duce the clinical application characteristics of PLGA microspheres,and review their application prospect in oral and maxillofacial tumors,temporomandibular joint diseases,periodontitis,caries and alveolar bone defects,so as to provide reference for their applica-tion in stomatology.
5.Breath-by-breath measurement of exhaled ammonia by acetone-modifier positive photoionization ion mobility spectrometry via online dilution and purging sampling
Lu WANG ; Dandan JIANG ; Lei HUA ; Chuang CHEN ; Dongming LI ; Weiguo WANG ; Yiqian XU ; Qimu YANG ; Haiyang LI ; Song LENG
Journal of Pharmaceutical Analysis 2023;13(4):412-420
Exhaled ammonia(NH3)is an essential noninvasive biomarker for disease diagnosis.In this study,an acetone-modifier positive photoionization ion mobility spectrometry(AM-PIMS)method was developed for accurate qualitative and quantitative analysis of exhaled NH3 with high selectivity and sensitivity.Acetone was introduced into the drift tube along with the drift gas as a modifier,and the characteristic NH3 product ion peak of(C3H6O)4NH4+(K0=1.45 cm2/V·s)was obtained through the ion-molecule reaction with acetone reactant ions(C3H6O)2H+(K0=1.87 cm2/V·s),which significantly increased the peak-to-peak resolution and improved the accuracy of exhaled NH3 qualitative identification.Moreover,the interference of high humidity and the memory effect of NH3 molecules were significantly reduced via online dilution and purging sampling,thus realizing breath-by-breath measurement.As a result,a wide quantitative range of 5.87-140.92 μmol/L with a response time of 40 ms was achieved,and the exhaled NH3 profile could be synchronized with the concentration curve of exhaled CO2.Finally,the analytical capacity of AM-PIMS was demonstrated by measuring the exhaled NH3 of healthy subjects,demon-strating its great potential for clinical disease diagnosis.
6.Investigation and analysis of 3021 cases of occupational hand injury in Xiaoshan District, Hangzhou
Xin LI ; Yiqian JIANG ; Wei DING
Chinese Journal of Industrial Hygiene and Occupational Diseases 2021;39(8):623-625
Objective:To investigate the causes and characteristics of occupational hand trauma in Xiaoshan District of Hangzhou, and to provide basis for formulating preventive measures and treatment.Methods:In July 2020, 3021 patients with occupational hand injury treated in Xiaoshan District from January 2017 to December 2019 were selected as the research object. The data of gender, age, injury month and time period of patients with occupational hand injury were collected, and their relationship with the causes of injury was analyzed.Results:Among 3021 patients with occupational hand trauma in Xiaoshan District, most of them were men (male to female ratio 2.05∶1) , and the proportion of injuries from 18 to 30 years old was relatively high (1508 cases, 49.92%) . The proportion of patients with cutting injury was high (1208 cases, 39.99%) , most of the injuries were at the distal end of metacarpophalangeal joint (2118 cases, 70.11%) , the proportion of injuries in summer was relatively high (1225 cases, 40.55%) . Most of them were injured from 8:00 to 16:00 every day (1684 cases, 55.74%) , of which 11:00-12:00 accounted for 29.46% (890 cases) . The excellent and good rate of functional recovery in all patients was 85.10% (2571/3021) . There were significant differences in the distribution of injury causes among patients with occupational hand trauma in different gender, age and seasons ( P<0.01) . Conclusion:In Xiaoshan District of Hangzhou, there are many cases of cutting injuries in patients with occupational hand injuries, with high incidence in summer and 11:00-12:00. Enterprises should strengthen the supervision and management of relevant time periods, especially to prevent the occurrence of cutting injuries.
7.Investigation and analysis of 3021 cases of occupational hand injury in Xiaoshan District, Hangzhou
Xin LI ; Yiqian JIANG ; Wei DING
Chinese Journal of Industrial Hygiene and Occupational Diseases 2021;39(8):623-625
Objective:To investigate the causes and characteristics of occupational hand trauma in Xiaoshan District of Hangzhou, and to provide basis for formulating preventive measures and treatment.Methods:In July 2020, 3021 patients with occupational hand injury treated in Xiaoshan District from January 2017 to December 2019 were selected as the research object. The data of gender, age, injury month and time period of patients with occupational hand injury were collected, and their relationship with the causes of injury was analyzed.Results:Among 3021 patients with occupational hand trauma in Xiaoshan District, most of them were men (male to female ratio 2.05∶1) , and the proportion of injuries from 18 to 30 years old was relatively high (1508 cases, 49.92%) . The proportion of patients with cutting injury was high (1208 cases, 39.99%) , most of the injuries were at the distal end of metacarpophalangeal joint (2118 cases, 70.11%) , the proportion of injuries in summer was relatively high (1225 cases, 40.55%) . Most of them were injured from 8:00 to 16:00 every day (1684 cases, 55.74%) , of which 11:00-12:00 accounted for 29.46% (890 cases) . The excellent and good rate of functional recovery in all patients was 85.10% (2571/3021) . There were significant differences in the distribution of injury causes among patients with occupational hand trauma in different gender, age and seasons ( P<0.01) . Conclusion:In Xiaoshan District of Hangzhou, there are many cases of cutting injuries in patients with occupational hand injuries, with high incidence in summer and 11:00-12:00. Enterprises should strengthen the supervision and management of relevant time periods, especially to prevent the occurrence of cutting injuries.
8.Homology and drug resistance gene analysis of Group B Streptococcus resistance induced by clindamycin
Jing GAO ; Yaping WANG ; Nanyan JIANG ; Yiqian PENG ; Lu LIU ; Ying ZHANG ; Yisheng CHEN ; Chunmei YING
Chinese Journal of Laboratory Medicine 2018;41(1):24-28
Objectives To investigate the homology and drug resistance gene of Group B Streptococcus ( GBS) Resistance induced by Clindamycin and provide basic data for clinical prevention and treatment of GBS Resistance infection induced by Clindamycin .Methods 921 strains of GBS were isolated at Obstetrics&Gynecology Hospital of Fudan University from January , 2014 to December , 2015.VITEK2-compact automatic bacterial susceptibility instrument was used to test their sensitivity to 7 antibacterial drugs.63 positive strains were chosen through D-inhibition zone trial which were drug resistant to Erythromycin and susceptible or intermediary to Clindamycin .The strain′s sequence type was identified by the method of multilocus sequence typing ( MLST typing ) .The drug resistance genes mefA & ermB to Erythromycin were detected by using PCR method .The analysis was carried out to reveal the relevance to drug resistance , multilocus sequence typing and drug resistance gene .Results Among 921 strains of GBS , the drug resistance rate was respectively 53.4% ( 492/921 strains ) to Erythromycin , 50.2% ( 462/921 strains) to Clindamycin, 34.7% ( 320/921 strains ) to Levofloxacin and 7.5% ( 69/921 strains ) to Nitrofurantoin.The drug resistance rate of Levofloxacin for 63 GBS strains was 27.0%(17/63 strains) and no drug resistant strain was found to Penicillin , Vancomycin & Nitrofurantoin.12 different ST types were involved in total, including a new ST type:ST1072.The most common ones were ST12 (30.1%) (20/63 strains) &ST19 (25.4%) (16/63 strains).The drug resistance rate of Levofloxacin with ST 19 (75.0%) (12/16 strains) was much higher than that of other ST types .The relevance ratio of mefA and ermB among 63 GBS strains was respectively 27.0%(17/63 strains) and 41.3%(26/63 strains).Conclusions The genetic diversity existed in Group B Streptococcus resistance induced by Clindamycin detected in this study . There was significant difference on drug resistance and relevant drug resistant genes among different ST types.
9.Effect of microRNA-29b on proliferation and migration of breast cancer cells and its molecular mechanism.
Yiqian JIANG ; Qingmin GUO ; Jianzhong GU ; Xiaoping XU ; Suhong AN ; Fang SU ; Yanhong BAO ; Changxin HUANG ; Xiaoxiang GUAN
Journal of Zhejiang University. Medical sciences 2017;46(4):349-356
OBJECTIVETo investigate the effects of microRNA(miRNA)-29b on the proliferation and migration of breast cancer cells and its molecular mechanism.
METHODSThe recombinant lentiviral expression vector (lenti-miRNA-29b) was constructed and transfected into 293T cells to obtain lentivirus particles that were used to infect breast cancer MCF-7 cells. Transfection efficiency of lenti-miRNA-29b in MCF-7 cells was identified by the expression of green fluorescent protein (GFP). The expression of miRNA-29b was detected by real-time PCR. The cell proliferation and migration were detected by CCK8 assay and Transwell assay, respectively. The bioinformatics softwares were used to predict and screen the downstream target genes regulated by miRNA-29b, which were verified by double luciferase reporter gene assay, RT-PCR and Western blot. The effects of screened target gene RTKN on the growth and migration of MCF-7 cells were verified by RTKN siRNA.
RESULTSRecombinant lentiviral expression vector of miRNA-29b were successfully constructed. About 90% and 60% of the breast cancer cells showed green fluorescence in lenti-miRNA-29b and lenti-miRNA-NC groups, respectively. The expression of miRNA-29b in lenti-miRNA-29b group increased significantly compared with the lenti-miRNA-NC group and blank control group (all<0.05); the proliferation and migration ability of MCF-7 cells significantly reduced compared with the control group (all<0.05). The screening with bioinformatics softwares found that the 3'UTR coding region RTKN had the binding site to miRNA-29b; the dual luciferase reporter gene assay showed that the luciferase activity decreased significantly after the MCF-7 cells were co-transfected with wild type RTKN-WT-3'UTR and miRNA-29b mimics report gene vector (<0.05). The RTKN proteins in MCF-7 cells were significantly decreased after transfection with siRNA-RTKN, and the proliferation and migration ability of MCF-7 cells were significantly reduced (all<0.05).
CONCLUSIONSMiRNA-29b can inhibit the proliferation, invasion and metastasis of breast cancer cells by inhibiting the expression of RTKN.
10. Preparation of chaperone-antigen peptide vaccine derived from human gastric cancer stem cells and its immune function
Yiqian JIANG ; Qingmin GUO ; Xiaoping XU ; Juncai LIANG ; Yiyang HE ; Suhong AN ; Fang SU ; Chaoyang LI ; Changxin HUANG
Chinese Journal of Oncology 2017;39(2):109-114
Objective:
To explore the method of extracting chaperone antigen peptide complexes from gastric cancer stem cells and its immune function.
Methods:
Gastric cancer stem cells and gastric cancer cells were screened by low temperature ultrasonic lysis. After salting out and dialysis, the lysate supernatant was processed with SDS-PAGE to analyze the expression of chaperone antigen peptide complexes, and then was separated and purified with CNBr-activated SepharoseTM 4B. Reverse high pressure liquid chromatography (HPLC), SDS-PAGE and Western blotting were used to analyze the purity and nature of the acquired albumen. Lymphocyte proliferation assay and lymphocytotoxicity assay were used to ditermine the immunological activity of the chaperone-antigen peptide complexes.
Results:
The chaperone antigen peptide complexes of gastric cancer stem cells were prepared and identified successfully, of which the main components were the antigen peptides of HSP60, HSP70, HSP90 and HSP110. 0.75 μg and 1.00 μg HSP70-antigen peptide and 1.00 μg HSP90-antigen peptide activated lymphocytes significantly. Their

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