1.A case-control study on gut microbiota diversity and species composition in obese/overweight children aged 2-6 years in Shanghai
Ping LIAO ; Qin YAN ; Yi ZHANG ; Xin HE ; Peiyun ZHU ; Jian QI ; Chazhen LIU ; Tong LIU ; Yan SHI ; Wenjing WANG
Journal of Environmental and Occupational Medicine 2024;41(3):243-250
Background Multiple studies have shown a close relationship between changes in gut microbiota composition and obesity, and research results are influenced by factors such as race and geographical location, but there are few studies on children. Objective To analyze the diversity of gut microbiota related to obesity in a population of 2-6 years old, observe the distribution characteristics and species differences of gut microbiota between obese/overweight and normal weight groups, and explore the association betweenobese/overweight and gut microbiota diversity. Methods Fecal samples were collected from 74 children aged 2-6 years in Shanghai, including 18 obese/overweight individuals, 6 males and 12 females (male to female ratio of 1∶2), and 56 normal weight individuals, 18 males and 38 females (male to female ratio is nearly 1∶2). The 16S rDNA was extracted from bacteria in fecal samples, followed by PCR amplification, cDNA construction, and high-throughput sequencing. Naive Bayes algorithm was used to perform taxonomic analysis (phylum, class, order, family, genus, species) and community diversity analysis (Sobs index, Shannon index, Shannoneven index, Coverage index, PD index, and principal co-ordinates analysis) on representative sequences and abundance of amplicon sequence variants (ASV). Wilcoxon rank sum test, P-value multiple test correction, and analysis of similarities were used to test differences between the two groups to obtain information on the distribution characteristics and species differences of intestinal microbiota in children. Results Seventy-four fecal samples were sequenced, and the sequencing results were subjected to quality control and filtering. A total of 4905306 optimized sequences were obtained, resulting in 1860 ASVs. The diversity data analysis of ASVs generated 889 species annotation results at 8 taxonomic levels. The alpha diversity analysis showed that the richness (Sobs index), diversity (Shannon index), evenness (Shannoneven index), and phylogenetic diversity (PD index) of fecal community of the obese/overweight children were increased compared to those of the normal weight children, but there were no statistical differences between the two groups (P>0.05). The beta diversity analysis showed that there was little difference in the composition of microbial species between the two groups, and no significant clustering separation was observed. The results of species composition analysis at phylum, order, family, and genus levels of 74 samples showed a consistent core microbiota structure in the two groups of gut microbiota, but there were differences in microbiota composition. The differences in microbial community composition between the two groups were manifested at the taxonomic levels of order, family, and genus, among which phylum Firmicutes, order Erysipelotrichales, family Erysipelatocyclostridiaceae, genus Erysipelotrichaceae_ UCG-003 and genus Catenibacterium were significantly enriched in the obese/overweight group and contributed significantly to the phenotypic difference of obese/overweight [linear discriminant analysis (LDA)=3.72, P<0.01; LDA=3.29, P<0.05). Phylum Proteobacteria, order Enterobacterales, family Enterobacteriaceae, genus unclassified was significantly enriched in the normal weight group and contributed significantly to the phenotypic difference of normal body weight (LDA=3.93, P<0.05). Conclusion The richness and diversity of gut microbiota in obese/overweight children aged 2-6 years in Shanghai are increased, but there is no difference compared to normal weight children. There is a difference in the composition of gut microbiota between the obese/overweight group and the normal weight group.
2. Effects of metabolites of eicosapentaenoic acid on promoting transdifferentiation of pancreatic OL cells into pancreatic β cells
Chao-Feng XING ; Min-Yi TANG ; Qi-Hua XU ; Shuai WANG ; Zong-Meng ZHANG ; Zi-Jian ZHAO ; Yun-Pin MU ; Fang-Hong LI
Chinese Pharmacological Bulletin 2024;40(1):31-38
Aim To investigate the role of metabolites of eicosapentaenoic acid (EPA) in promoting the transdifferentiation of pancreatic α cells to β cells. Methods Male C57BL/6J mice were injected intraperitoneally with 60 mg/kg streptozocin (STZ) for five consecutive days to establish a type 1 diabetes (T1DM) mouse model. After two weeks, they were randomly divided into model groups and 97% EPA diet intervention group, 75% fish oil (50% EPA +25% DHA) diet intervention group, and random blood glucose was detected every week; after the model expired, the regeneration of pancreatic β cells in mouse pancreas was observed by immunofluorescence staining. The islets of mice (obtained by crossing GCG
3. The molecular mechanism of spleen-strengthening and moisture-nourishing liver prescription in treatment of acute-on-chronic liver failure based on network pharmacology and experimental verification
Qi HUANG ; Wen-Feng MA ; Zhi-Yi HAN ; Jia-Ling SUN ; Wei ZHANG ; Xin-Feng SUN ; Jian -Ping CHEN ; Xiao-Zhou ZHOU ; Qi HUANG ; Wen-Feng MA ; Zhi-Yi HAN ; Jia-Ling SUN ; Wei ZHANG ; Xin-Feng SUN ; Xiao-Zhou ZHOU ; Jing LI ; Xiao-Zhou ZHOU ; Jian -Ping CHEN
Chinese Pharmacological Bulletin 2024;40(3):557-564
To explore the mechanism of spleen- were obtained for the treatment of acute-on-chronic livstrengthening and moisture-nourishing liver prescription er failure, and 244 intersecting target genes and 7 core (JPLSYGF) in the treatment of acute-on-chronic liver target genes were screened. Molecular docking showed failure using network pharmacology and the molecular that the core target genes AKT1, SRC, VEGFA, docking. Methods Relying on TCMSP and Gene- STAT3 , EGFR, MAPK3 , HRAS had good affinity with Cards and other databases, the relevant targets of JPL- quercetin, the main active component in the JPLSYGF in the treatment of acute-on-chronic liver failure SYGF, and had strong binding activity. In addition, in were obtained. String and Cytoscape were used to con- vivo tests verified that the JPLSYGF could reduce the struct PPI networks of targets, core targets were expression of HRAS, EGFR, STAT3 , SRC, and VEGscreened out, and DAVID was used for GO function FA, to delay the progression of acute-on-chronic liver annotation and KEGG pathway enrichment analysis. failure. Conclusions JPLSYGF may act on core tar- The main active ingredients of the traditional Chinese gets such as HRAS, EGFR, STAT3, SRC, VEGFA medicine compound formula for JPLSYGF were select- and so on, to achieve the effect of treating acute-oned with a bioavailability OB value of =Э 30% and a chronic liver failure. drug-like DL^O. 18 as the screening conditions, and.
4.Role of Guiqi Yiyuan ointment combined with cisplatin in the treatment of Lewis lung cancer based on PI3K/Akt/mTOR signal pathway
Chao YUAN ; Si-Qi KONG ; Jian-Qing LIANG ; Yi ZHANG ; Rong HU ; Yue ZHANG ; Yu LIU ; Jin-Tian LI
The Chinese Journal of Clinical Pharmacology 2024;40(10):1424-1428
Objective To observe the inhibitory effect of Guiqi Yiyuan ointment on tumor growth in mice with Lewis lung cancer,and to explore the molecular mechanism of Guiqi Yiyuan ointment combined with cisplatin through phosphoinositide 3-kinase/protein kinase B/mammalian rapamycin target protein(PI3K/Akt/mTOR)signal pathway.Methods Sixty C57BL/6 mice were randomly divided into 6 groups with 10 mice in each group.Except for the blank group(0.9%NaCl),Lewis lung cancer-bearing mice were randomly divided into model group(0.9%NaCl),control group(0.9%NaCl,cisplatin 5 mg·kg-1)and low,medium,high dose experimental groups(Guiqi Yiyuan ointment 1.6,3.3,6.6 g·kg-1,cisplatin 5 mg·kg-1).Flow cytometry was used to detect bone marrow-derived suppressor cells(MDSCs);the expression of related proteins in tumor tissues was detected by Western blot.Results The tumor inhibition rates in control group and low,medium,high dose experimental groups were(39.87±4.45)%,(45.74±14.97)%,(57.78±4.70)%and(69.82±11.05)%.The proportion of MDSCs in bone marrow of in blank group,model group,control group and low,medium,high dose experimental groups were(36.13±1.08)%,(68.63±2.94)%,(58.93±2.02)%,(58.00±1.50)%,(50.93±5.06)%and(43.07±2.41)%.The protein expressions of p-PI3K/PI3K in model group,control group and low,medium and high experimental groups were 0.97±0.03,0.77±0.02,0.72±0.01,0.68±0.03 and 0.53±0.02;PTEN were 0.21±0.07,0.65±0.07,0.74±0.06,0.99±0.13,1.11±0.13;p-Akt/Akt were 1.01±0.02,0.82±0.02,0.77±0.00,0.72±0.03 and 0.52±0.04;p-mTOR/mTOR were 1.01±0.01,0.76±0.05,0.69±0.07,0.59±0.06 and 0.47±0.06.There were significant differences between low,medium,high experimental groups and control group(all P<0.05).Conclusion Guiqi Yiyuan ointment combined with cisplatin can significantly improve the quality of life and inhibit tumor growth in mice.The mechanism may be the inhibition of PI3K/Akt/mTOR signal pathway and the enhancement of tumor cell apoptosis and autophagy.
5.Effects of Platycodon grandiflorum Bai powder in the treatment non-small cell lung cancer rats
Chao YUAN ; Jin-Tian LI ; Jian-Qing LIANG ; Yi ZHANG ; Si-Qi KONG ; Rong HU ; Yue ZHANG ; Yu LIU
The Chinese Journal of Clinical Pharmacology 2024;40(11):1608-1612
Objective To observe the effects of traditional Chinese medicine compound Platycodon grandiflorum Bai powder on the growth of subcutaneously implanted tumor and the expression of B-cell lymphoma-2(Bcl-2),Bcl-2 associated X protein(Bax),cysteinyl aspartate specific proteinase(caspase)-3 and caspase-9 in subcutaneously implanted tumor of Lewis lung cancer mice.Methods The model of transplanted tumor of Lewis lung cancer in mice was established.Seventy SPF male C57BL/6 mice were randomly divided into blank group,model group,low dose experimental group,medium dose experimental group,high dose experimental group,control group and combined group.Blank group and model group were given 0.9%NaCl 0.2 mL by gavage;control group was given 0.9%NaCl by gavage and 25 mg·kg-1cisplatin intraperitoneally;high,medium,low dose experimental groups were given 193,96,48 mg·kg-1·d-1 Platycodon grandiflorum Bai powder 0.2 mL by gavage,respectively;combined group was given 96 mg·kg-1·d-1 Platycodon grandiflorum Bai powder 0.2 mL by gavage,and 25 mg·kg-1 cisplatin intraperitoneally,once every other day.The myelogenous suppressor cells(MDSCs)of mouse bone marrow were detected by flow cytometry,and the expressions of Bel-2,Bax,caspase-3 and caspase-9 in tumor cells were detected by immunofluorescence.Results The percentage of MDSCs in bone marrow of mice in blank group,model group,low dose experimental,medium,high dose experimental group,control group and combination group were(32.50±2.76)%,(63.13±3.14)%,(48.43±2.23)%,(42.53±1.28)%,(32.93±3.56)%,(51.30±4.25)%and(19.90±6.21)%,respectively.The fluorescence intensities of Bax in model group,low dose experimental group,medium dose experimental group,high dose experimental group,control group and combination group were 10.42±0.68,12.40±1.23,15.14±0.65,22.95±1.76,27.18±1.62 and 31.61±1.28;Bel-2 were 36.85±0.80,33.92±4.20,28.88±1.01,20.04±2.21,15.69±2.36 and 6.05±0.73;caspase-3 were 5.28±0.44,7.63±0.55,9.66±0.85,14.73±1.18,17.95±1.29 and 22.92±1.95;caspase-9 were 9.48±0.90,11.57±0.72,13.45±0.93,15.73±1.44,19.20±0.96 and 23.21±1.51.There were significant differences between medium,high dose experimental groups and model group(all P<0.05),and there were significant differences between combined group and control group(all P<0.05).Conclusion Platycodon grandiflorum Bai powder can up-regulating the expression of Bax,caspase-3 and caspase-9,down-regulating the expression of Bel-2,inhibiting MDSCs,promoting tumor cell apoptosis and inhibiting tumor growth.
6.Effects of sodium acetate on lowering uric acid and renal protection in mice with hyperuricemic nephropathy
Xue-Man LIN ; Shi-Qi ZHONG ; Yong-Mei LI ; Xiao-Yi QIN ; He-Yang JIANG ; Jia-Xin ZHOU ; Jian-Xin PANG ; Ting WU
The Chinese Journal of Clinical Pharmacology 2024;40(15):2222-2226
Objective To investigate the renal protective effect and mechanism of sodium acetate(Ace)on hyperuricemic nephropathy(HN)in mice.Methods Uric acid nephropathy mice model was prepared by intraperitoneal injection of potassium oxonate combined with adenine gavage.Mice were divided into blank control group(0.9%NaCl+0.5%carboxymethyl cellulose sodium),Ace group(200 mmol·L-1 Ace+0.5%carboxymethyl cellulose sodium),model group(0.9%NaCl+350 mg·kg-1 potassium oxonate+70 mg·kg-1 adenine),and experimental group(based on model group with additional 200 mmol·L-1 Ace).Serum and urine uric acid(UA)and serum creatinine(SCr)levels were observed in each group.Real-time fluorescence quantitative reverse transcription-polymerase chain reaction(qRT-PCR)was used to detect the expression levels of kidney injury molecule-1(Kim-1)and anti-aging gene Klotho,renal fibrosis markers Collagen Ⅰ and Fibronectin,intestinal inflammation-related factors interleukin-1 β(IL-1 β),and mRNA expression levels of tight junction proteins Zo-1.Results The serum UA levels of blank control group,Ace group,model group,and experimental group mice were(259.52±24.40),(227.71±35.91),(604.06±73.55),and(496.24±30.16)μmol·L-1,respectively;SCr levels were(16.85±0.40),(16.18±0.94),(22.38±1.56),and(19.78±1.43)μmol·L-1;Kim-1 mRNA relative expression levels were 1.04±0.25,1.17±0.28,13.00±2.87,and 4.24±3.92;Klotho mRNA relative expression levels were 1.04±0.15,1.02±0.18,0.43±0.12,and 0.69±0.12;Collagen Ⅰ mRNA relative expression levels were 1.05±0.15,1.02±0.18,3.19±1.09,and 1.61±0.55;Fibronectin mRNA relative expression levels were 1.07±0.18,1.02±0.25,7.86±2.40,and 3.34±2.10;intestinal IL-1β mRNA relative expression levels were 1.00±0.01,1.01±0.03,2.55±0.63,and 1.21±0.28;intestinal Zo-1 mRNA relative expression levels were 1.00±0.07,1.07±0.09,0.54±0.20,and 0.92±0.17.The above indicators in blank control group compared with model group,and experimental group compared with model group,all showed statistically significant differences(P<0.05,P<0.01,P<0.001).Conclusion Sodium acetate can effectively reduce UA levels in HN mice,significantly improve renal injury and fibrosis,and its mechanism may be related to the improvement of intestinal inflammatory response and up-regulation of intestinal Zo-1/Occuludin pathway to reduce intestinal mucosal permeability.
7.Naphthoquinone and carboxylic acid derivatives from the fungus Pleosporales sp.
Si-yuan QI ; Li-hua ZHANG ; Yi-nan HAO ; You-cai HU ; Jian BAI
Acta Pharmaceutica Sinica 2024;59(5):1327-1333
Three new compounds, including a naphthoquinone, a reduced naphthoquinone derivative naphthalenone, and a tricarboxylic acid, along with five known naphthalenone derivatives were isolated from ethyl acetate extract of rice fermentation products of the fungus
8.EIT-based study of lung ventilation and perfusion regional distribution and ventilation/perfusion matching in different body positions
Ti-Xin HAN ; Yi-Fan LIU ; Meng DAI ; Pu WANG ; Jian-An YE ; Zhi-Bo ZHAO ; Zhan-Qi ZHAO ; Feng FU
Chinese Medical Equipment Journal 2024;45(5):1-7
Objective To compare regional pulmonary ventilation and perfusion and ventilation/perfusion(V/Q)matching in different body positions using electrical impedance tomography(EIT).Methods Ten healthy experimental pigs were selected to collect their EIT lung ventilation and perfusion data in supine,prone,left lateral and right lateral positions.The EIT data underwent analysis and image reconstruction using MATLAB R2022b and EIDORS v3.9.The effective regions with ventilation and perfusion were determined and the V/Q matching regions were computed with the maximum pixel value 20%as the threshold.Comparisons were carried out over the V/Q matching indexes including V/Q match%,dead space%and shunt%and ventilation and perfusion distribution in regions of interest(ROIs)including ROI1,ROI2,ROI3 and ROI4 in different body positions.Results The differences in V/Q match%,dead space%and shunt%of the experimental animals in varied positions were not statistically significant(P>0.05).The regional distribution of pulmonary ventilation and perfusion changed in different positions,and the regional distributions differed in ROIl,ROI2 and ROI3 for ventilation(P<0.05)and in ROI1 and ROI2 for perfusion(P<0.05).The ventilation and perfusion regions were distributed consistently with the gravity-dependent areas in supine and prone positions whereas conversely in the right and left lateral positions.Conclusion The V/Q matching indexes of one subject have high test consistency in different body positions;gravity-dependent areas varied with the changes of the body positions,which affected the distribution of pulmonary ventilation and perfusion regions;EIT can be used for measuring the changed pulmonary ventilation and perfusion due to different positions and determining the influences of position changes on pulmonary ventilation and perfusion and V/Q matching.[Chinese Medical Equipment Journal,2024,45(5):1-7]
9.Detection of o-Phthalaldehyde by Two-dimensional Photonic Crystal Hydrogel Sensor
Jian-Wei XIN ; Yi-Fei WANG ; Zi-Hui MENG ; Yu-Qi ZHANG ; Peng-Fei LIU
Chinese Journal of Analytical Chemistry 2024;52(4):523-530
o-Phthalaldehyde(OPA)is a new type of chemical disinfectant widely used in medical institutions.The development of new efficient and convenient detection platforms or methods for OPA is of great significance.In this work,in two-dimensional photonic crystal(2DPC)hydrogel,a responsive 2DPC hydrogel was prepared by functionalizing the hydrogel with ethylenediamine(EDA)and embedding amino groups.The amino group on the polymer chain of 2DPC hydrogel reacted with OPA,and with the increase of OPA concentration,the crosslinking density of the hydrogel also increased,resulting in the volume phase transition of the hydrogel,e.g.,shrinkage phenomenon.In the meantime,the spacing of 2DPC microspheres gradually decreased,while the diameter of Debye diffraction ring gradually increased.The results showed that the change of particle size spacing had a good linear relationship with logarithm of concentration of OPA in the range of 101?106 nmol/L,with the detection limit of 0.21 nmol/L(3σ/k).Therefore,the amino functionalized photonic crystal hydrogel sensor could realize the quantitative detection of OPA.The method was simple with low cost,ease to operate and use.Then the practicability of this hydrogel sensor for real sample was verified in the diluted clinical disinfectant.The recoveries of OPA in the diluted disinfectant were 100%?103%,with a relative standard deviations of 1.8%?5.5%.The results proved that 2DPC hydrogel sensor could be used for detection of OPA in disinfectant used for clinical endoscopes and other instruments.
10.Influences and mechanism of XIST on proliferation and extracellular matrix synthesis of nucleus pulposus cells in rats with intervertebral disc degeneration
Gao-Chen WU ; Jin-Peng CHEN ; Fan-Jian MENG ; Lu-Lu WANG ; Yi-Qi MIAO
Medical Journal of Chinese People's Liberation Army 2024;49(7):823-831
Objective To investigate the influences and mechanism of X chromosome inactivation specific transcript(XIST)on the proliferation and extracellular matrix synthesis of nucleus pulposus cells in rats with intervertebral disc degeneration(IDD).Methods SD rat intervertebral disc nucleus pulposus cells were isolated and cultured in vitro,and then randomly divided cells into control group,model group,XIST knockdown group,empty plasmid group,and XIST knockdown+miR-132-3p knockdown group.Except for control group,the cells in other groups were induced by interleukin(IL)-1β to establish IDD models.After nucleus pulposus cells were grouped and treated,the expressions of XIST and miR-132-3p in the nucleus pulposus cells of rats were detected by qRT-PCR;the proliferation of intervertebral disc nucleus pulposus cells was detected by MTS method and EdU staining;ELISA was used to measure the levels of inflammatory factors IL-6 and IL-18 in the intervertebral disc nucleus pulposus cells;Immunoblotting was used to detect the expression of extracellular matrix labeled proteins Collagen Ⅱ and Aggrecan in rat nucleus pulposus cells.The targeted regulation of miR-132-3p by XIST in rat nucleus pulposus cells was detected by dual-luciferase reporter gene assay.IDD rat models were established by intramuscular injection of IL-1β and divided into sham operation group,model group,XIST knockdown group,no-load plasmid group,XIST knockdown+miR-132-3p knockdown group,with 12 rats in each group.After treatment in each group,the expressions of XIST and miR-132-3p in intervertebral disc tissues were detected by qRT-PCR;TUNEL staining was used to detect apoptosis of nucleus pulposus cells in intervertebral disc tissue of rats;the morphology of intervertebral disc cartilage was observed by saffron O staining;serum levels of inflammatory factors IL-6 and IL-18 were determined by ELISA;the expressions of Collagen Ⅱ and Aggrecan in intervertebral disc tissues were detected by Western blotting.Results Compared with control group(cells)/sham operation group(rats),XIST expression in intervertebral disc tissue and nucleus pulposus cells,apoptosis rate of intervertebral disc nucleus pulposus cells,levels of inflammatory factors IL-6 and IL-18 in intervertebral disc nucleus pulposus cells culture medium and serum were increased in model group(P<0.05),the activity and proliferation rate of nucleus pulposus cells and the expressions of miR-132-3p,Collagen Ⅱ and Aggrecan protein in nucleus pulposus cells and intervertebral disc tissues decreased(P<0.05);compared with model group,the apoptosis rate of nucleus pulposus cells,the levels of inflammatory factors IL-6 and IL-18 in intervertebral disc nucleus pulposus cells culture medium and serum,and the expression of XIST in nucleus pulposus cells and intervertebral disc tissues of rats in XIST knockdown group decreased(P<0.05),the activity and proliferation rate of nucleus pulposus cells and the expressions of miR-132-3p,Collagen Ⅱ and Aggrecan protein in nucleus pulposus cells and intervertebral disc tissues increased(P<0.05).Down-regulation of miR-132-3p attenuates the ameliorative effect of knockdown XIST on IL-1β-induced intervertebral disc injury and cartilage matrix loss.XIST was able to target and down-regulate the expression of miR-132-3p in rat nucleus pulposus cells.Conclusion Knockdown of XIST can inhibit inflammation by up-regulating miR-132-3p,thereby inhibiting the apoptosis of IDD nucleus pulposus cells and promoting their proliferation and extracellular matrix synthesis.

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