Table of Content

Volume 27 Issue 2
26 February 2022

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Prolactinic effects and molecular mechanisms of total sterone from Echinops latifolius Tausch on the milk deficient model rats

WANG Xiao, XUE Qiuyun, HUANG Yurong, CHENG Chenglong, HUANG Yuting, CHANG Jun, YIN Qun, DU Mingsong, MIAO Chenggui

2022, 27(2):  121-128.  doi:10.12092/j.issn.1009-2501.2022.02.001

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AIM: To investigate the effect of an effective component total sterone (TSR) of Echinops latifolius Tausch, the main component of a Chinese patent medicine Cuiru Keli (national drug standard WS3-413 (Z-085)-2003 (Z), on lactation and its possible mechanism. METHODS: After mating between male and female SD rats, 60 female rats were randomly divided into normal control group, model group, TSR low-dose and high-dose groups and prolactin granule positive control group, with 12 female rats in each group and 8 newborn rats in each nest. In addition to the normal control group, the rats in each group were intraperitoneally injected with levodopa 2 mg/kg once a day for 7 days from the second day of delivery. The rats in the normal control group were given normal saline by gavage once a day for 14 days. From the beginning of self-sufficiency, the single lactation of the female rats was measured every day until the 14th day, and then the female rats in each group were killed. Pathological HE staining was used to observe the morphological changes of mammary gland tissue in each group. ELISA was used to detect the levels of serum prolactin (PRL) and 5-hydroxytryptamine (5-HT). Immunohistochemistry was used to detect the distribution of PRL in mammary gland tissue of each group. Furthermore, Real-time qPCR was used to detect the expression of milk protein, milk fat related genes β-casein, FAS, ACC and the expression of canonical Wnt signaling pathway related genes β-catenin, c-Myc, CCND1, SFRP4, DNMT1, MeCP2 in mammary gland of each group. RESULTS: Both low and high dose TSR could significantly increase the single lactation volume, improve the pathological morphology of mammary gland, and increase the serum levels of PRL and 5-HT. TSR increased the distribution of PRL and up-regulated the expression of milk protein, milk fat related genes β-casein, FAS, ACC and canonical Wnt signaling pathway related genes β-catenin, c-Myc, CCND1, SFRP4, DNMT1, MeCP2.CONCLUSION: TSR can significantly promote lactation in lactation deficient rats, and its mechanism may be related to promoting the release of PRL and 5-HT in serum, increasing the distribution of PRL in mammary gland, up-regulating the milk protein and milk fat related genes and activating the canonical Wnt signal.

Effects of liquorice extract on cardiac fibroblasts fibrosis induced by TGF-β1 

NIU Pilian, FAN Yongxin, LU Furui, ZHOU Xuezhang, BAI Mingsheng

2022, 27(2):  129-135.  doi:10.12092/j.issn.1009-2501.2022.02.002

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AIM: To investigate the effect of liquorice extract on TGF-β1-induced myocardial fibroblast (CFs) fibrosis.  METHODS: 10 ng/mL TGF-β1 induced CFs to establish myocardial fibrosis cell model. Fibrotic cells were treated with liquorice extract and the cell activity was detected by MTT assay. CCK-8 was used to detect the effect of liquorice extract on CFs proliferation. The expression of smooth muscle actin (α-SMA) was detected by immunofluorescence. Western blot was used to detect TGF-β1/Smad signaling pathway related proteins and p-Smad2, p-Smad3 expression levels. The mRNA expression levels of Smad2, Smad3 and Smad4 were detected by RT-PCR. RESULTS: Compared with the control group, there were statistically significant differences in cell activity (P<0.05). The cell proliferation rate of glycyrrhiza uralensis extract groups was significantly lower than that of TGF-β1 group (P<0.05). The expression levels of α-SMA and TGF-β1/Smad signaling pathway related proteins in 100 μg/mL liquorice extract were significantly lower than those in TGF-β1 group (P<0.05). CONCLUSION: Glycyrrhiza extract can improve the occurrence and development of TGF-β1-induced myocardial fibrosis, and its mechanism maybe related to the inhibition of TGF-β1/Smad signaling pathway.

Effects of Shenqi fuzheng injection on low-glucose-mediated immunosuppressive microenvironment and its mechanism of action

MA Wangbo, Ma Yue, FAN Fangtian, ZHANG Yuhan, CHANG Jingwen, ZHOU Zhihua

2022, 27(2):  136-143.  doi:10.12092/j.issn.1009-2501.2022.02.003

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AIM: To investigate the effect of Shenqi fuzheng injection (SFI) on tumor immunity and its preliminary molecular mechanism. METHODS: The animal model of low glucose tumor microenvironment was established by B16-PKM2-OE; the level of interleukin-2(IL-2) and interferon-γ(IFN-γ), CD40L and transforming growth factor-β1(TGF-β1) were detected by ELISA kit; the expressions of glucose transporter-1 (Glut-1) and key enzymes of glycolysis ( HK, PFK and PK ) in CD4+T cells were detected by Western blot. RESULTS: The results of the kit showed that SFI could increase IL-2, IFN-γ and CD40L, and decreased TGF-β1 in tumor tissues. Western blot assay showed that SFI could promote the expression of Glut-1 protein on the surface of CD4+T cell membrane, and up-regulated the expression of key enzymes of glycolysis (HK, PFK1 and PK) in a dose-dependent manner. CONCLUSION: SFI can increase the expression levels of IL-2, IFN-γ, CD40L and decrease the expression level of TGF-β1 in tumor tissues. Also, it increases the number of activated CD4+T cells in tumor tissue and promotes the glycolysis of CD4+T cells, indicating that SFI can improve the tumor immune microenvironment.

Effects of ABT-737 on the growth and angiogenesis of ovarian cancer cells in the co-culture of TAMs and SKOV3 cells

YAO Yao, XIA Min, HOU Cong, HE Airong

2022, 27(2):  144-153.  doi:10.12092/j.issn.1009-2501.2022.02.004

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AIM: To explore the effect of Bcl-2 small molecule inhibitor ABT-737 on the growth and angiogenesis mimicry of SKOV3 cells in a co-culture system of Tumour-associated macrophages (TAMs) and human ovarian cancer cells SKOV3. METHODS: PMA and IL-4 was used to induce THP-1 cells into TAMs cells in vitro; MTT method was used to detect the cell survival rate of SKOV3 cells after 24 hours of treatment with different concentrations of ABT-737 culture medium; a co-culture system of SKOV3 cells and TAMs cells was established; the experimental groups were divided into control group, SKOV3+ABT-737 group (containing 5.0 μmol/L ABT-737 cultured cells), TAMs+SKOV3 group (SKOV3 cells co-cultured with TAMs cells), TAMs+SKOV3+ABT-737 group (SKOV3 cells Co-cultured with TAMs cells, and added ABT-737 containing 5.0 μmol/L), cells after 24 h was collected, MTT method was used to detect cell survival rate, EdU staining for cell proliferation, ranswell chamber experiment for cell migration and invasion, Flowcytometry for cell apoptosis, the vascular mimicry experiment for the ability of cells to form blood vessels, Western blot for the expression of vascular endothelial growth factor (VEGF), matrix metalloproteinase-2 (MMP-2) and MMP-9 in cells. RESULTS: THP-1 cells were successfully induced for TAMs cells; the survival rate of SKOV3 cells decreased under the action of ABT-737 (P<0.01); compared with the control group, the survival rate of SKOV3 cells in the SKOV3+ABT-737 group decreased, the number of EdU-labeled positive cells decreased, the number of cell migration and invasion also decreased, the rate of apoptosis increased, and the duct branches decreased, The protein expression of VEGF, MMP-2, MMP-9 decreased (P<0.01); Compared with the TAMs+SKOV3 group, the cell survival rate of the TAMs+SKOV3+ABT-737 group decreased, the number of EdU-labeled positive cells and the number of cell migration and invasion also decreased, the apoptosis rate increased, and the duct branches decreased. At the same time, the protein expression of VEGF, MMP-2, MMP-9 decreased (P<0.01). CONCLUSION: ABT-737 can inhibit SKOV3 cell proliferation, metastasis, apoptosis and angiogenesis in a co-culture system, and affect tumor progression.

Preventive and therapeutic effects of the Kiwi fruit essence (unsaturated fatty acid of actinidia chinesis planch seed oil) on radiation-induced lung injury rats 

HE Liyang, WEI Wenjie, YIN Qin, LIU Lijing, ZHONG Boying, LIU Cungen, ZHOU Meiling, HE Jianbin

2022, 27(2):  154-162.  doi:10.12092/j.issn.1009-2501.2022.02.005

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AIM: To observe the effects of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), myeloperoxidase (MPO) and transforming growth factor-β1 (TGF-β1) of radiation-induced lung injury rats by Kiwi fruit essence (unsaturated fatty acid of actinidia chinesis planch seed oil).  METHODS: According to random number table, 90 Sprague-Dawley rats were divided into the control group, model group, 60 mg/kg unsaturated fatty acid of actinidia chinesis planch seed oil intervention group, 120 mg/kg unsaturated fatty acid of actinidia chinesis planch seed oil intervention group, 240 mg/kg unsaturated fatty acid of actinidia chinesis planch seed oil intervention group, 18 animals were included in each group. Except for the control group, rats in the remaining groups were constructed by 6MV-X-ray 18Gy full chest radiation, one week before modeling, the rats in the last 3 groups were given (60, 120, 240) mg/kg unsaturated fatty acid of actinidia chinesis planch seed oil. The first two groups were given 0.9% sodium chloride solution by gavage, once a day in each rat. 14 days, 28 days, and 56 days after radiation, the rats were randomly sacrificed and their chests were cut, and ave lung tissue was saved. HE and Masson staining were performed to observe the pathological changes, and the content of SOD, GSH-Px, MPO was determined. The expression of TGF-β1 was analyzed by immunohistochemical staining. RESULTS: Compared with the model group, (60, 120, 240) mg/kg unsaturated fatty acid of actinidia chinesis planch seed oil significantly reduced the degree of lung alveolitis and radiation pulmonary fibrosis, reduced the content of hydroxyproline (HYP), MPO, increased the antioxidant enzymes SOD and GSH-Px content, down-regulated the expression of TGF-β1.There were significant differences in the above-mentioned indicators among the intervention groups of (60, 120, 240) mg/kg unsaturated fatty acid of actinidia chinesis planch seed oil group, and it was positively correlated with dosage. CONCLUSION: Unsaturated fatty acid of actinidia chinesis planch seed oil has a preventive effect on radiation-induced lung injury, and its mechanism may be related to the reduction of oxidative stress damage and down-regulation of TGF-β1 expression.

Effects of miR-195-5p on ox-LDL-induced vascular endothelial cell injury through regulating FBXW7

LIN Naping, WU Yutang, HUANG Defen, XU Chaoxiang

2022, 27(2):  163-170.  doi:10.12092/j.issn.1009-2501.2022.02.006

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AIM: To investigate the effect of miR-195-5p on ox-LDL-induced vascular endothelial cell injury and its mechanism.  METHODS: The expression of miR-195-5p in HUVECs cells induced by ox-LDL was detected by RT-PCR. Cell proliferation, the expression of oxidative stress associated factors (MDA and LDH) and apoptosis were detected by CCK-8, ELISA and flow cytometry, respectively. The potential targets of miR-195-5p were determined by dual luciferase reporter assay. The expression of target protein in ox-LDL-induced HUVECs cells and the relationship between miR-195-5p and FBXW7 expression were detected by Western blotting. RESULTS: The expression of miR-195-5p in ox-LDL-induced HUVECs cells was significantly higher than that in normal HUVECs cells. Subsequently, miR-195-5p silencing in ox-LDL-induced HUVECs cells effectively promoted the proliferation of HUVECs cells, whereas suppressed the expression of oxidative stress associated factors (MDA and LDH) and apoptosis level. Luciferase reporter assay and Western blot results showed that miR-195-5p could directly target FBXW7 protein gene and negatively regulate its expression. Finally, miR-195-5p modulated the proliferation, oxidative stress and apoptosis of HUVECs cells induced by ox-LDL by regulating the expression of FBXW7. CONCLUSION: miR-195-5p is highly expressed in HUVECs cells induced by ox-LDL. In addition, miR-195-5p can aggravate ox-LDL-induced cytotoxicity, oxidative stress and apoptosis of HUVECs cells by inhibiting the expression of FBXW7.

Correlation between organic cation transporter gene polymorphisms and the toxicities and clinical response of oxaliplatin

CHEN Jiayin, WANG Li, WANG Lijun, TONG Gangling, MA Jie, CHEN Xijing, LU Yang

2022, 27(2):  171-177.  doi:10.12092/j.issn.1009-2501.2022.02.007

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AIM: To investigate the relationship between genotypes of rs628031, rs650284, rs683369 of SLC22A1 gene and the toxicities and clinical response of oxaliplatin in patients with colorectal cancer.  METHODS: A total of 72 patients diagnosed as colorectal cancer during January 2018 to June 2018 were selected and all patients received oxaliplatin treatment. Their peripheral venous blood was collected and genotyping was conducted by using SNaPshot. The toxicities including gastrointestinal toxicity, hematological toxicity and peripheral neurotoxicity were evaluated according to the Common Terminology Criteria Adverse Events (CTCAE) Version 5.0. Clinical response was evaluated according to the Response Evaluation Criteria in Solid Tumors (RECIST Version 1.1). RESULTS: The results of Chi-test showed that different genotypes of SLC22A1 SNP sites rs628031 and rs683369 may be related to the toxicities and clinical response of oxaliplatin significantly. Specifically, when compared with the patients with GG type of rs628031, the patients with the GA or AA type had a lower incidence of grade 3 nausea and vomiting (P=0.017) and may also be less responsive to efficacy (P=0.008). When compared with the patients with CC type of rs683369, the patients with the GC or GG type had a lower incidence of grade 3 nausea and vomiting (P=0.002) and may also be less responsive to efficacy (P=0.014).CONCLUSION: The polymorphisms of SLC22A1 gene are closely related to the toxicities and clinical response of oxaliplatin in patients with colorectal cancer, which may be helpful for improving clinical treatment.

Clinical trials of inhalation in pediatric population in China

ZHU Lili, ZHU Xiuxiu, WANG Wenjun, ZHANG Hailin, LIN Li

2022, 27(2):  178-183.  doi:10.12092/j.issn.1009-2501.2022.02.008

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AIM: To investigate the characteristics of clinical trials of inhalation in pediatric population in China. METHODS: The pediatric clinical trials of inhaled drugs in China registered on the www.Chinadrugtrials.org.cn and Clinical Trials in USA respectively until November 20, 2021 were reviewed. The characteristics of pediatric clinical trials of inhaled drugs including the clinical trial phases, drug indications and classificatio etc. were analyzed. RESULTS: There were 21 pediatric clinical trials of inhaled drugs registered on the www.Chinadrugtrials.org.cn, accounted for 8.9%(21/235) of inhalation clinical trials in all populations. 47.6% of them were generic drugs, mainly focusing on expectorants for Phlegm symptoms and inhaled preparations for asthma, which accounting for 71.4%(15/21) . There were 34 pediatric clinical trials of inhaled drugs registered on the Clinical Trials in USA, the drug indications of which were mainly asthma and anesthesia, accounting for 76.5%(26/34). CONCLUSION: The pediatric clinical trials of inhalations in China started later, and the total number is small compared to adults, mainly focusing on generic drugs. We should pay attention to the research and development of new inhalation drugs, standardizing and promoting the clinical trials of inhaled drugs in pediatric population actively.

Effects of alfentanil on tracheal intubation during tonsillectomy in children: A randomized double-blind study

LI Jiajia, CHEN Mengmeng, WANG Ruixian, HUANG Mengmeng, LI Jun, SHANGGUAN Wangning

2022, 27(2):  184-189.  doi:10.12092/j.issn.1009-2501.2022.02.009

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AIM: To compare the effects of different doses of alfentanil on tracheal intubation conditions, hemodynamic parameters and recovery quality in children undergoing tonsillectomy.  METHODS: Ninety children undergoing tonsillectomy were randomly divided into 3 groups, and received alfentanil 20 μg/kg (A20 group), 40 μg/kg (A40 group) and 60 μg/kg (A60 group) for anesthesiainduction respectively, 30 cases in each group. The remaining anesthesia induction and maintenance protocols were the same. The Helbo-Hansen scores of the three groups were evaluated, and the MAP and HR before anesthesia induction (T0), before tracheal intubation (T1), immediately after tracheal intubation (T2), and 1 min after intubation (T3) as well as the recovery time of spontaneous breathing, eye opening time, extubated time, agitation score in PACU, and adverse drug reactions were recorded. RESULTS: Compared with A20 group, the total values of Helbo-Hansen score and cough scores in group A40 and A60 were lower (P<0.05). Compared with T0, the MAP at T1-T3 were decreased in group A40 and A60, and HR increased at T2 and T3 in group A20 while HR slowed down at T1 in group A40, and at T1-T3 in group A60 (P<0.05). Compared with A20 group, children in group A40 had lower MAP and slower HR at T1-T3, while those in group A60 had lower MAP and slower HR at T1-T3 (P<0.05). The recovery time of spontaneous breathe and extubated time were prolonged in group A60 (P<0.05). CONCLUSION: During the anesthesia induction period of tonsillectomy in children, both afentanil 40 μg/kg or 60 μg/kg combined with propofol 3 mg/kg and rocuronium 0.3 mg/kg can provide satisfactory intubation condition, while the vital signs are more stable during anesthesia induction in afentanil 40 μg/kg group and rapid extubation after operation can be achieved.

Design and evaluation of clinical trials of COVID-19 vaccine and monoclonal neutralizing antibody

LI Shanshan, GU Jingwen, ZHANG Jing, YANG Haijing, LIU Wei, YU Yiqi, ZHANG Wenhong

2022, 27(2):  190-197.  doi:10.12092/j.issn.1009-2501.2022.02.010

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COVID-19 pandemic has put a huge burden on public health and global economy. Vaccines play an important role in controlling virus transmission and reducing mortality. While monoclonal virus neutralizing antibodies can reduce the viral load, improve symptoms, and prevent the aggravation of the disease from hospitalization. Now hundreds of clinical trials of COVID-19 vaccine and monoclonal neutralizing antibody are in progress. The vaccine focuses on disease prevention, while the neutralizing antibody focuses on disease treatment. There are quite many differences between the two kinds of clinical trials by following different technical guidelines, research purpose, trial design, implementation and outcome assessment. Therefore, it is necessary to summarize the similarities and differences between the clinical trials for the reference of new drug research and development as well as clinical researchers.

Recent progress of mitophagy in hepatic insulin resistance

QUAN Haiyan, JIANG Xing, HE Lu

2022, 27(2):  198-204.  doi:10.12092/j.issn.1009-2501.2022.02.011

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Metabolic syndrome, characterized by centralobesity, hypertension, bycentralobesity, hypertension, and hyperlipidemia, increases the incidence and mortality of cardiovascular disease, type 2 diabetes, nonalcoholic fatty liver disease, and other metabolic diseases. It is well known that insulin resistance, especially hepatic insulin resistance, is a risk factor for metabolic syndrome. Current research has shown that the accumulation of hepatic fatty acid can cause hepatic insulin resistance through increased gluconeogenesis, lipogenesis, chronic inflammation, oxidative stress and endoplasmic reticulum stress, and impaired insulin signal pathway. Mitochondria are the major sites of fatty acid β-oxidation, which is the major degradation mechanism of fatty acids. Mitochondrial dysfunction has been shown to be involved in the development of hepatic fatty acid-induced hepatic insulin resistance. Mitochondrial autophagy (mitophagy), a catabolic process, selectively degrades damaged mitochondria to reverse mitochondrial dysfunction and preserve mitochondrial dynamics and function. Therefore, mitophagy can promote mitochondrial fatty acid oxidation to inhibit hepatic fatty acid accumulation and improve hepatic insulin resistance. Here, we review advances in our understanding of the relationship between mitophagy and hepatic insulin resistance. Additionally, we also highlight the potential value of mitophagy in the treatment of hepatic insulin resistance and metabolic syndrome.

Association of Lin28 with tumor, cardiovascular disease and diabetes

LV Xiaohan, LIN Rong

2022, 27(2):  205-211.  doi:10.12092/j.issn.1009-2501.2022.02.012

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Lin28 is a highly conserved RNA-binding protein, including Lin28a and Lin28b. Lin28 is involved in many physiological processes such as cell differentiation, growth and metabolism. In recent years, it has been found that Lin28 is closely related to the pathogenesis and development of numerous major diseases. The relationships between Lin28 and related diseases such as tumor, cardiovascular disease and diabetes are reviewed in this article.

Pharmacology and clinical evaluation of vericiguat in the treatment of heart failure

LIU Ping, QIU Bo, WU Huizhen

2022, 27(2):  212-218.  doi:10.12092/j.issn.1009-2501.2022.02.013

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Vericiguat is a soluble guanylate cyclase stimulator, acting on nitric oxide-soluble guanylate cyclase-cyclic guanosine monophosphate pathway. Vericiguat can improve the sensitivity soluble guanylate cyclase sensitivity to nitric oxide, stimulate soluble guanylate cyclase without relying on nitric oxide, leading to increased formation of cyclic guanosine monophosphate , which results in multi-dimensional protection effects for the heart. It provides a new therapeutic approach for patients with heart failure. This review provides an overview of mechanism, preclinical studies, pharmacokinetics, clinical efficacy, drug-drug interactions and limitations of vericiguat.

Research progress of short-chain fatty acids in alleviating organ ischemia-reperfusion injury

LV Xingjiao, LENG Yufang, HAN Xiaoxia, HOU Xiaoyu, CAO Xuefen, Janvier NIBARUTA, LIU Yongqiang

2022, 27(2):  219-226.  doi:10.12092/j.issn.1009-2501.2022.02.014

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Short-chain fatty acids (SCFAs) are organic acids with no more than 6 carbon atoms produced during the anaerobic fermentation of dietary fiber in the intestinal tract, which can regulate intestinal flora, repair intestinal mucosal barrier, and reduce intestinal injury.Ischemia-reperfusion injury (IRI) is the main cause of various diseases, and the pathological mechanisms involved are intricate, among which inflammation, oxidative stress, apoptosis and autophagy are the most common. According to current studies, SCFAs can affect the occurrence and development of IRI in various organs by regulating different cell signal transduction. In this paper, the role and mechanism of SCFAs in alleviating tissue and organ ischemia-reperfusion injury were preliminarily summarized, providing theoretical reference for clinical prevention and treatment of IRI.

Ferroptosis regulatory signaling pathway and its research progress in related diseases

ZHANG Liang, LIAO Yongqun, XIA Qinchuan, ZHOU Shitong, LI Xiaoli

2022, 27(2):  227-234.  doi:10.12092/j.issn.1009-2501.2022.02.015

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Ferroptosis is an iron-dependent novel type of programmed cell death. The main features of ferroptosis include lipid reactive oxygen accumulation, iron accumulation and lipid peroxidation. The main mechanisms and signal pathways of ferroptosis are complex and closely related to cystine/glutamate antiporter system, glutathione peroxidase 4, ferroptosis suppressor protein 1, and dihydroorotate dehydrogenase. This review summarizes the current regulatory mechanisms of ferroptosis and discusses the research progress of ferroptosis in tumors, non-alcoholic fatty liver disease, Parkinson's disease, and congestive heart failure.

Research progress on hydrogen sulfide in tissue repair and regeneration

WU Junchi, YU Nannan, ZHAO Chengcheng, LIU Wenhui, LUO Zhiying, CAI Hualin, LIU Yiping

2022, 27(2):  235-240.  doi:10.12092/j.issn.1009-2501.2022.02.016

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Hydrogen sulfide (H2S), a colorless gas with a strong odor of rotten eggs, is an important biological signaling molecule, which is produced in vivo through enzyme-catalyzed and non-enzymatic pathways. Physiological levels of H2S have systemic antioxidant, anti-inflammatory, regulating apoptosis, dilating blood vessels, lowering blood pressure, neuroprotective effects, and prevent bone damage. H2S plays an important role in tissue repair, which is of great value for clinical treatments. In this paper, the common repair and regeneration functions and mechanisms of hydrogen sulfide in tissue damage will be reviewed, which will be helpful to understand the role of H2S in vivo and the development of new drugs.

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2022, 27(2):  241. 

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