1.Effect of survivin shRNA on chemotherapy resistance in GBC-SD cells
Hanbin SHEN ; Yaohui WU ; Haocheng LONG ; Qichang ZHENG ; Jianping GONG
Chinese Journal of Hepatobiliary Surgery 2010;16(1):43-47
Objective To investingate the effect of survivin shRNA on chemotherapy resistance in GBC-SD cells. Methods They were divided into three groups of GBC-SD, GBC-SD/EGFP and GBC-SD/survivin. MTT assay was used to detect cell viability in the 3groups. mRNA and protein of survivin were tested by RT-PCR and Western blot. Then the cells were treated with proper construction DDP. The cell survival rate and IC_(50) were determined with MTT, cell apoptosis detected by FACS and the alteration of nucleus observed by TUNEL. Meanwhile, caspase-3 activity was determined using the colorimetric method. Results Cell viability was reduced remarkably in GBC-SD/survivin and survivin expression was decreased obviously. After being treated with DDP, cell survival rate and IC_(50) were decreased obviously in GBC-SD/survivin, apoptotic rate elevated remarkably compared with other groups. There were brownly nucleuses in three groups. Caspase-3 activity increased first and then decreased, but it exceeded in GBC-SD/survivin than that in other two groups. Conclusion The survivin shRNA can down-regulate the expression of survivin in GBC-SD cells remarkably and improve the sensibility to chemotherapy.
2.Dynamic evolution mechanism and treatment ideas of colorectal"inflammation-mediated carcinogenesis"based on the"latent pathogens trapped in the intestinal collaterals"theory
Yunze LIU ; Haocheng ZHENG ; Yuan LI ; Xia DING
Journal of Beijing University of Traditional Chinese Medicine 2024;47(10):1343-1348
The risk of colorectal cancer is substantially higher in patients with long-term colorectal inflammation than in the general population.Prolonged inflammation is an essential factor that triggers colorectal cancer.The dynamic pathological evolution process of the classic"inflammation-mediated carcinogenesis"in the colorectum is proctocolitis→dysplasia→cancer.Traditional Chinese medicine lacks a systematic consensus on the pathogenesis of colorectal"inflammation-mediated carcinogenesis".This article proposes the theory of"pathogenic factors lurk intestinal collaterals"to explain the development law of pathogenesis in the dynamic evolution of colorectal"inflammation-mediated carcinogenesis".Internal and external factors can trigger the movement of the latent pathogenic factors,thereby damaging the intestinal tissues,when latent pathogenic factors are hidden in the intestinal collaterals,and the healthy qi is unable to expel them.The prolonged course of the disease further weakens the healthy qi,allowing the latent pathogenic factors to accumulate in the intestinal collaterals,intertwine with phlegm-dampness and blood stasis,accelerate accumulation,and lead to cancer."Latent pathogens trapped in the intestinal collaterals"is the core pathogenesis during inflammation-mediated carcinogenesis.Thus,to prevent colorectal inflammation-mediated carcinogenesis,consideration should be given to the principle of driving away pathogenic factors and dredging the collaterals in clinical practice.The theory regarding"latent pathogens trapped in the intestinal collaterals"can provide a theoretical reference for syndrome differentiation and treating colorectal inflammation-mediated carcinogenesis and offer novel ideas for clinical treatment.
3.Retrospective study on the impact of penile corpus cavernosum injection test on pe-nile vascular function
Yan CHEN ; Kuangmeng LI ; Kai HONG ; Shudong ZHANG ; Jianxing CHENG ; Zhongjie ZHENG ; Wenhao TANG ; Lianming ZHAO ; Haitao ZHANG ; Hui JIANG ; Haocheng LIN
Journal of Peking University(Health Sciences) 2024;56(4):680-686
Objective:To investigate the impact of age,various hormonal levels,and biochemical markers on penile cavernous body vascular function in patients with erectile dysfunction(ED).Me-thods:A retrospective analysis of clinical data from male patients with ED who underwent color duplex Doppler ultrasonography(CDDU)and intracavernosal injection test(ICI)at the Reproductive Medicine Center of Peking University Third Hospital from January 2020 to August 2023.Data were managed and processed using SPSS 29.0,and a multivariable Logistic regression analysis was conducted.Results:A total of 700 ED patients were included,with 380 showing negative ICI results and 320 positive.In the study,84 patients had a peak systolic velocity(PSV)<25 cm/s,while 616 had PSV ≥ 25 cm/s;202 patients had end-diastolic velocity(EDV)>5 cm/s,and 498 had EDV ≤5 cm/s.264 patients had ab-normal PSV and/or EDV results,and 436 had normal results for both.Patients with vascular ED had sig-nificantly lower estrogen levels(t=-3.546,P<0.001),lower testosterone levels(t=-2.089,P=0.037),and a higher rate of hyperglycemia(x2=12.772,P=0.002)compared with those with non-vascular ED.The patients with arterial ED were older(t=3.953,P<0.001),had a higher rate of hyperglycemia(x2=9.518,P=0.009),and a higher estrogen/testosterone ratio(t=2.330,P=0.020)compared with those with non-arterial ED.The patients with mixed arteriovenous ED had higher age(t=3.567,P<0.001),lower testosterone levels(t=-2.288,P=0.022),a higher rate of hyperglycemia(x2=12.877,P=0.002),and a larger estrogen/testosterone ratio(t=2.096,P=0.037)compared with those with normal findings.Multifactorial Logistic regression analysis indicated that higher levels of estrogen were a protective factor for vascular ED(OR=1.009,95%CI:1.004-1.014),and glucose 7.0 mmol/L was a risk factor(OR=0.381,95%CI:0.219-0.661).Older age was a risk factor for arte-rial ED(OR=0.960,95%CI:0.938-0.982).Additionally,older age(OR=0.976,95%CI:0.958-0.993)and glucose levels of 5.6-6.9 mmol/L(OR=0.591,95%CI:0.399-0.876)were also risk fac-tors for mixed arterio-venous ED.Conclusion:Hyperglycemia and aging may impair penile cavernous body vascular function,while higher levels of estrogen may have a protective effect on it.
4.Traditional Chinese Medicine Treats Chronic Atrophic Gastritis via Multiple Pathways
Sici WANG ; Yuan LI ; Ping LI ; Haocheng ZHENG ; Weiyu CHEN ; Xia DING
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(19):193-202
ObjectiveTo mine the compatibility rules of patented traditional Chinese medicine (TCM) compound prescriptions for treating chronic atrophic gastritis (CAG) by systems pharmacology and molecular docking methods, and predict the targets and molecular mechanisms of Chinese medicinals with different efficacy in the treatment of CAG. MethodThe TCM compound prescriptions for treating CAG were extracted from the patent system of the China National Intellectual Property Administration. The active components and targets of the prescriptions were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP), Traditional Chinese Medicine Integrative Database (TCMID), and UniProt. The candidate targets and pathways of CAG were obtained from GeneCards, DisGeNet, Online Mendelian Inheritance in Man (OMIM), MalaCards, Kyoto Encyclopedia of Genes and Genomes (KEGG) and Reactome. The gene ontology (GO) functional annotation and KEGG pathway enrichment were realized by R Studio 4.1.2. STRING11.0 was employed to build the protein-protein interaction (PPI) network, and AutoDock Vina 4.2.6 was used for the docking between key targets and components. ResultA total of 228 TCM compound prescriptions for treating CAG were extracted. The medicinals used in these prescriptions mainly had warm or cold nature, bitter or sweet taste, tropism to the spleen, stomach, and liver meridians, and the efficacy of tonifying Qi, regulating Qi movement, clearing heat, and activating and toniying blood. The prescriptions mainly treated CAG via p53, mitogen-activated protein kinase (MAPK), phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt), forkhead box protein O (FoxO), vascular endothelial growth factor (VEGF), and hypoxia-inducible factor-1 (HIF-1) signaling pathways. Molecular docking results confirmed that the active components in the prescriptions had docking activities with key receptor proteins. ConclusionThis study preliminarily analyzed the compatibility rules of TCM compound prescriptions in the treatment of CAG. The medicinals with different efficacy treat CAG by regulating cell proliferation, apoptosis, and oxidative stress response, preventing carcinogen production, promoting gastric acid secretion, and improving local microcirculation in a multi-target, multi-pathway, multi-link manner. The findings facilitate the research on the TCM treatment of CAG.
5.Computational fluid dynamics analysis of influence of different pipe structures on gas mixing uniformity
Jin’ge ZHENG ; Weiya CHENG ; Chenxiao WANG ; Guizhen HAO ; Weifu LIU ; Haocheng GUO ; Yulun ZHAO ; Ling CHEN
Chinese Journal of Radiological Health 2022;31(2):172-180
Objective Tostudy the influence of pipe structures on the mixing uniformity of airborne effluents from nuclear power plant chimneys. Methods We used the computational fluid dynamics (CFD) method to simulate the velocity distribution and gas mixing in long straight pipes (I type) with square section and circular section, 90° single-bend pipes (L type) with square section and circular section, and 90° double-bend pipes (S type and U type) with square section and circular section. Results For the long straight pipe, due to the lack of flow disturbance caused by structural changes, the mixing effect was not good; when the pipe section was circular, it might take mixing distance 20 times the hydraulic diameter to achieve the uniformity index required by the relevant standard; for the square pipe, the distance might be longer. In the single bend pipe with square section, the velocity uniformity was improved more greatly after the bend, and the tracer gas met the mixing uniformity at a shorter distance (11 times the hydraulic diameter), as compared with the single bend pipe with circular section. For the S-type double-bend pipe, the tracer gas appeared uniformly mixed after a distance 6 times the hydraulic diameter in the square pipe, and 7 times the hydraulic diameter in the circular pipe. For the U-type double-bend pipe, the gas in the square pipe also achieved uniform mixing ata shorter distance downstream, and the airflow showed greater disturbance when passing through the bend. Conclusion The CFD method can make an accurate prediction for the change patterns of gas mixing uniformity in pipes with different structures, and can partially replace physical experiments to study the factors affecting the mixing uniformity of airborne effluents from the chimney of nuclear power plants.
6.Dayuanyin Regulates TLR/MAPK/NF-κB Pathway for Preventing and Treating Acute Lung Injury Induced by H1N1 Infection
Chengze LI ; Fuhao CHU ; Yuan LI ; Yunze LIU ; Haocheng ZHENG ; Sici WANG ; Yixiao GU ; Wanhong ZHU ; Ruoshi ZHANG ; Xingjian SONG ; Cong GAI ; Xia DING
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(18):52-60
ObjectiveTo investigate the therapeutic effect of Dayuanyin on acute lung injury induced by H1N1 infection and decipher the potential mechanism. MethodThe constituents in Dayuanyin were analyzed by ultra-high performance liquid chromatography-quadrupole-exactive orbitrap mass spectrometry (UHPLC-Q-Exactive Orbitrap MS). Forty-eight female BALB/c mice were randomized into normal, model, oseltamivir (19.5 mg·kg-1), and low-, medium-, and high-dose (2.73, 5.46, 10.92 g·kg-1) Dayuanyin groups. The normal and model groups were administrated with deionized water by gavage, and the other groups were administrated with the corresponding drugs by gavage. On day 3 of drug administration, the normal group received nasal inhalation of normal saline, and the other groups were inoculated intranasally with A/RP/8/34 (H1N1) for the modeling of influenza virus infection. Mice were administrated with drugs continuously for 7 days and weighed daily. Sampling was performed 12 h after the last administration, and the lung tissue was weighed to calculate the lung index. Hematoxylin-eosin staining was performed to observe the pathological and morphological changes of the lung tissue and bronchi. The cytometric bead array (CBA) was used to measure the serum levels of interferon-gamma (IFN-γ), C-X-C motif ligand 1 (CXCL1), tumor necrosis factor-alpha (TNF-α), chemokine ligand 2 (CCL2), interleukin-12p70 (IL-12p70), chemokine ligand 5 (CCL5), interleukin-1β (IL-1β), chemokine (C-X-C motif) ligand 10 (CXCL10), granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin-10 (IL-10), interferon-beta (IFN-β), interferon-alpha (IFN-α), and interleukin-6 (IL-6). According to the results of mass spectrometry and network pharmacology, we analyzed the mechanism of Dayuanyin in treating acute lung injury caused by H1N1. The protein levels of extracellular signal-regulated kinase 1/2 (ERK1/2), p38 mitogen-activated protein kinase (p38 MAPK), nuclear factor-kappa B (NF-κB), and their phosphorylated forms were determined by Western blot. The mRNA levels of myeloid differentiation factor 88 (MyD88), Toll-like receptor 3 (TLR3), Toll-like receptor 7 (TLR7), and Toll-like receptor 8 (TLR8) in the lung tissue were measured by Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR). ResultA total of 57 compounds, including paeoniflorin and baicalein, were detected in Dayuanyin. Compared with the normal group, the model group showed decreased body weight (P<0.01), lung edema and hemorrhage, increased lung index (P<0.01), and elevated levels of IFN-γ, IL-12p70, CCL5, IL-1β, CXCL10, GM-CSF, IFN-β, and IL-6 (P<0.01). Compared with the model group, Dayuanyin attenuated alveolar wall thickening, capillary congestion, and immune cell infiltration, reduced the alterations in body weight and lung index (P<0.01), and down-regulated the protein levels of IFN-γ, IL-12p70, CCL5, IL-1β, CXCL10, GM-CSF, IFN-β, and IL-6 (P<0.01). A total of 57 key genes were predicted by network pharmacological analysis, of which the MAPK signaling pathway was the main target signaling pathway. Compared with the normal group, the model group showed up-regulation in the protein levels of phosphorylation (p)-ERK1/2, p-p38 MAPK, and p-NF-κB (P<0.01) and the mRNA levels of TLR7, TLR8, MyD88, and TLR3 (P<0.05, P<0.01). Compared with the model group, Dayuanyin lowered the phosphorylation levels of ERK1/2, p38 MAPK, and NF-κB p65 in a dose-dependent manner (P<0.01) and down-regulated the mRNA levels of TLR3, TLR7, TLR8, and MyD88 (P<0.01). ConclusionDayuanyin can prevent and control H1N1 infection-induced acute lung injury by inhibiting the TLR/MAPK/NF-κB signaling pathway.