Table of Content

Volume 25 Issue 4
26 April 2020

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Expression of long non-coding RNA LINC00844 in hepatocellular carcinoma and its inhibiting effect on cell proliferation and migration

HUANG Kang, LI Ling, YE Qifa, PENG Guizhu

2020, 25(4):  366-372.  doi:10.12092/j.issn.1009-2501.2020.04.003

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AIM: To explore the expression level of long non-coding RNA(lncRNA) LINC00844 and its effect on hepatocellular carcinoma (HCC). METHODS: The expression of LINC00844 was detected by real-time quantitative PCR (qRT-PCR) in 12 pairs of HCC tissues and paired paracancerous tissues. The LINC00844 group (LV-LINC00844) and the control group (LV-NC) in HepG2 and HCCLM9 cell lines were constructed by lentivirus carrying LINC00844 gene and lentivirus without LINC00844 gene. The effects of over-expressed LINC00844 on proliferation and migration of HepG2 and HCCLM9 cell lines were detected by cell counting kit-8 (CCK-8) assay and transwell assay, respectively.RESULTS:Among 12 pairs of clinical samples, the expression of LINC00844 in 11 hepatocellular carcinoma tissues was significantly lower than that in paired paracancerous tissues detected by qRT-PCR (P<0.01). Over-expressed HepG2 and HCCLM9 cell lines were successfully constructed. Furthermore, in vitro experiments demonstrated that enhanced expression LINC00844 inhibted the proliferation and migration of HepG2 and HCCLM9 (P<0.01). CONCLUSION: LINC00844 can inhibit the proliferation and migration in HepG2 and HCCLM9 cell lines. LINC008444 may be a potential promising prognostic biomarker and therapeutic target for HCC.

Research of peripheral mechanism of opioid receptor on testosterone secretion

NAN Haihan, ZHOU Jiangping, HUANG Jianjian, HAO Xinrui, CUI Yanhua, CHEN Xingchi, LIN Han

2020, 25(4):  373-379.  doi:10.12092/j.issn.1009-2501.2020.04.004

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AIM: To investigate the effect of opioid μ receptor on testosterone and its peripheral mechanism.  METHODS: Levels of testosterone and testosterone synthetase mRNA in testis plasma and testis homogenate, IGF1 protein and mRNA expression in testis homogenate were detected by intratesticular morphine injection in vivo animal model and treatment of specific μ receptor antagonists and agonists in vitro cultured testis. RESULTS:Serum testosterone level was decreased after testis was injected with morphine ［(722.11±121.02) vs. (346.91±75.31) pg/mL; t=7.898, P=0.001］. Testis homogenate testosterone level ［(133.61±16.82) vs. (66.56±14.96) pg/mg protein; t=8.941, P=0.001］ and expression of testosterone synthetase were decreased. Testis IGF1 level ［(7.93±2.13) vs. (2.54±0.74) ng/mg protein, t=7.155, P=0.001］ and IGF1 mRNA ［(3.22±1.12) vs. (1.66±0.55), t=3.751, P=0.001］ expression were decreased in intratesticular morphine injection model. Testosterone level in culture media, IGF1 and expression of testosterone synthetase were decreased after testis was injected with DAMGO in vitro model. And testosterone level, IGF1 and expression of testosterone synthetase were increased after testis was injected with CTOP.CONCLUSION: Morphine acts on opioid μ receptor on the surface of testicular Sertoli cells and inhibits the synthesis of IGF1. It decreases expression of testosterone synthetase Scarb1, Star, 3βHsd, Cyp17α1 and 17βHsd from testicular Leydig cells. which leads to a decrease in testosterone synthesis.

Inhibiting RNA methylase NSUN2 reduce the invasion and migration of gastric cancer cell

SHEN Cheng, YANG Nianzhao, WANG Minghai, WU Lisheng, ZHU Zhiqiang

2020, 25(4):  380-386.  doi:10.12092/j.issn.1009-2501.2020.04.005

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AIM: To investigate the effect of RNA methylase NSUN2 on the invasion and migration ability of gastric cancer cells, and to explore its mechanism. METHODS: The slow-transfected virus that inhibited the NSUN2 gene and the no-load virus that was negative control were transfected into gastric cancer SGC7901 cell lines and MGC803 cell lines, respectively. Real-time quantitative PCR and Western blot were used to detect the mRNA and protein expression of NSUN2 in two groups, TRANSWELL and scratch experiments were used to study the effects of NSUN2 on cell migration and invasion, Western blot was used to detect the Epithelial-Mesenchymal Transition (EMT) molecules in the two groups of cells Marker (E-Cadherin, N-Cadherin) protein expression. RESULTS:Compared with the control group, the expression of RNA methylase NSUN2 mRNA and protein in the interference group decreased, the number of transmembrane cells decreased, the migration distance decreased, the expression of N-Cadherin protein decreased, and the expression of E-Cadherin protein increased. All differences have statistical significance.CONCLUSION: Inhibition of NSUN2 gene expression can reduce the invasion and migration of gastric cancer cell lines SGC7901 and MGC803, and the mechanism is to inhibit the EMT process.

Forsythiaside B inhibits the proliferation activity of HeLa cells though transcription factor NF-κB

ZHANG Zhe, LYU Yu, HUANG Panhao

2020, 25(4):  387-392.  doi:10.12092/j.issn.1009-2501.2020.04.006

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AIM: To study the effects of forsythiaside B on HeLa cells proliferation by inhibiting NF-κB through p21/Cyclin E/CDK2 signaling pathway. METHODS: The cell activity of HeLa cells was observed by MTT assay after 24 h, 48 h and 72 h treatment with different concentration (1, 2, 4 μmol/L) of forsythiaside B. The clone formation of HeLa cells was evaluated with the condition of different concentrations of forsythiaside B. The expression of transcription factor NF-κB (p-p65 and p65) was determined, and the expression of p21, Cyclin E and CDK2 were detected as well.RESULTS:Forsythioside B inhibited the clone formation of HeLa cells in concentration dependent manner, and the inhibitory effect on HeLa cells' proliferation by forsythioside B was in both time and concentration dependent manner. Forsythioside B up-regulated p21 protein expression and down-regulated p-p65, Cyclin E and CDK2 protein expression. CONCLUSION:Forsythioside B inhibits the transcription factor NF-κB and affects the p21/Cyclin E/CDK2 signaling pathway, thus inhibiting the proliferation of HeLa cells.

Preparation of genistein combined polysaccharide liposomes and its inhibitory effects on prostate cancer

XU Shenghua, GAO Zhuo, WANG Jianfeng

2020, 25(4):  393-400.  doi:10.12092/j.issn.1009-2501.2020.04.007

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AIM: To prepare genistein combined polysaccharide (GCP) liposomes and to evaluate the potential value of GCP liposomes for the treatment of prostate cancer by in vivo and in vitro experiments, and ultimately to promote the application of nanotechnology in prostate cancer chemotherapy. METHODS: GCP liposomes were prepared by ethanol injection. The anti-tumor effect of GCP liposomes was verified in vitro and in vivo by cell proliferation assay and cell cycle analysis, as well as establishing a tumor model of prostate cancer.RESULTS:GCP liposomes enhanced the inhibitory effect of GCP on androgen-sensitive LNCaP cell proliferation and the apoptosis of androgen-sensitive LNCaP cells induced by GCP. Also, GCP liposomes improved the inhibitory effect of GCP on tumor growth in tumor-bearing mice. CONCLUSION: GCP liposomes have better anti-tumor effect on androgen-sensitive human prostate cancer cells than GCP.

Mulberry polysaccharides improves the chemotherapy of liver cancer ascites tumor-bearing mice by regulating the PI3K/AKT/mTOR pathway

ZHAO Miaohui, ZHOU Jiani, QI Yong

2020, 25(4):  401-407.  doi:10.12092/j.issn.1009-2501.2020.04.008

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AIM: To study the synergistic and attenuating effects of mulberry polysaccharides on the chemotherapy of liver cancer ascites tumor-bearing mice by regulating the PI3K/AKT/mTOR pathway. METHODS: Ninety SPF male Kunming mice were randomly divided into normal group, model group, cyclophosphamide group, mulberry polysaccharide group and mulberry polysaccharide + cyclophosphamide group. Liver cancer ascites tumor-bearing mice were prepared, and each administration group was given 30 mg/kg of cyclophosphamide or (and) 200 mg/kg of mulberry polysaccharide and administered orally. The normal group and the model group were administrated with 10 mL/kg saline. The tumor suppression rate, liver, spleen and other indexes, tumor tissue VEGF and inflammatory factor content, and PI3K/AKT/mTOR pathway-related protein expression were observed in mice.RESULTS:Cyclophosphamide group, mulberry polysaccharide group and mulberry polysaccharide + cyclophosphamide group mice body weight, tumor mass, tumor tissue VEGF, TNF-α, IL-6, PI3K, AKT and mTOR phosphorylation levels were lower than the model group, and the level of IL-1β was higher than the model group; mulberry polysaccharide + cyclophosphamide group mice were observed with body weight, tumor inhibition rate, tumor tissue VEGF, TNF-α, IL-6, PI3K, AKT and The mTOR phosphorylation level higher than the mulberry polysaccharide group and cyclophosphamide group, and the tumor mass and IL-1β level were lower than the mulberry polysaccharide group and cyclophosphamide group (P<0.05). CONCLUSION: Morus alba polysaccharide combined with cyclophosphamide can effectively inhibit tumor proliferation of liver cancer ascites tumor-bearing mice and exert attenuating effect. The mechanism may be related to the down-regulation of PI3K /AKT/mTOR pathway expression.

Application of model-informed drug development approach in dose optimization: Insights from the modification of nivolumab dosage regimen

LI Jian, YAN Fang, YANG Jinbo, WANG Yuzhu

2020, 25(4):  408-412.  doi:10.12092/j.issn.1009-2501.2020.04.009

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Nivolumab is a humanized monoclonal antibody that blocks the programmed death 1 (PD-1) receptor. The initially recommended dose was 3 mg/kg every 2 weeks (Q2W) when nivolumab was firstly approved by the US FDA in 2014. A flat dose of 240 mg Q2W was approved in 2016, and subsequently 480 mg Q4W in 2018. Model-informed drug development (MIDD) approach was applied for the analyses of drug exposure, safety and efficacy based on the data from existed clinical studies, and the model-informed analyses became the key evidences supporting approval of new dosage regimens. Relevant studies and regulatory considerations related to the nivolumab dose selection and subsequent modification were reviewed in this paper. The application of MIDD approach in dose optimization is additionally discussed.

Response surface model to optimize the concentration range of propofol and remifentanil

YANG Lu, WEI Bin, ZHANG Liping, BI Shanshan, LU Wei

2020, 25(4):  413-420.  doi:10.12092/j.issn.1009-2501.2020.04.010

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AIM: To investigate the pharmacodynamic interactions between propofol and remifentanil by response surface methodology. The response surface for inhibition of somatic, hemodynamic responses to laryngoscopy and response surface of cardiovascular side effects were combined to identify the optimal combination of propofol and remifentanil. METHODS: Seventy ASA I-II patients were grouped randomly and parallelly. The patients received target-controlled infusion (TCI) of propofol (0-9 μg/mL) at a fixed concentration, then remifentanil was administrated with TCI device from 0 to 10 ng/mL in a stepwise ascending fashion. After reaching pseudo-steady-state drug levels, the somatic and hemodynamic responses to laryngoscopy and cardiovascular side effects were observed. The pharmacodynamic interactions were investigated by response surface methodology. RESULTS:Response surface models revealed significant synergy between propofol and remifentanil. Imax were 2.89, 5.02 and 2.55 for the somatic and hemodynamic responses to laryngoscopy and cardiovascular side effects. Based on the results of response surface method, the optimal dose range of propofol and remifentanil was constructed.CONCLUSION: Response surface method can analyze the pharmacodynamic interactions qualitatively and quantitatively and provide the optimal dose range.

Protection of subjects' legal rights, welfare and safety in clinical trials of COVID-19

ZHANG Zhengfu, LI Meng, WANG Jianan

2020, 25(4):  421-425.  doi:10.12092/j.issn.1009-2501.2020.04.011

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Since December 2019, COVID-19 has been found in Wuhan, Hubei Province of China, which has spread in many countries and regions around the world. A large number of clinical trials have been launched in China aiming to find safe and effective drugs and treatments.The protection of subjects' legal rights, welfare and safety should be superior to the consideration of scientific and social benefits. Therefore, all parties involved in clinical trials should take corresponding measures to protect subjects' legal rights, welfare and safety. This paper discussed about the protection of subjects' legal rights, welfare and safety in clinical trials of COVID-19 from all aspects of the clinical trials, aiming to provide reference for all parties involved in clinical trials and basic ideas for the protection of subjects' legal rights, welfare and safety in clinical trials of emergencies.

Evaluation of the effects of resveratrol on the activity human cytochrome P450 through combined probe method

WU Qiaoyu, YUAN Hong, LU Yao

2020, 25(4):  426-432.  doi:10.12092/j.issn.1009-2501.2020.04.012

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AIM: To evaluate the effects of resveratrol on five human cytochrome enzyme P450 subtypes, i.e. CYP1A2, CYP2D6, CYP2C9, CYP3A4, and CYP2C19, through a combined probe method. METHODS: We conducted a randomized controlled study in which 26 healthy male volunteers were recruited and were randomized into the resveratrol group and the placebo group. Volunteers were given oral placebo /resveratrol at a dose of 1 g each time, once daily for 14 days. Then, they took five oral probe drugs, caffeine (for CYP1A2), losartan (for CYP2D6), omeprazole (for CYP2C19), dextromethorphan (for CYP2C9), and midazolam (for CYP3A4). Blood samples at different times were collected to detect the concentration of the three probes, i.e. midazolam, caffeine and omeprazole; 0-8 h urine samples were collected to measure the metabolic rate of losartan and dextromethorphan. RESULTS:Compared with the placebo group, the AUC0-12, AUC0-∞, Cmax of caffeine in the resveratrol group was increased by 32.10% (P<0.001), 71.52% (P<0.001), and 10.62%, respectively (P=0.004), and T1/2 was also increased significantly (P=0.002). The AUC0-12, AUC0-∞, T1/2 and Cmax of omeprazole or midazolam in the resveratrol group showed no significant change (P>0.05) as compared with the placebo group. The 8 h urinary DTM/DT ratio was increased by 79.91% (P=0.003) and 8 h urinary losartan/E-3174 ratio was increased by 531.34% (P<0.001).CONCLUSION: Resveratrol significantly inhibited the enzymatic activities of CYP1A2, CYP2D6 and CYP2C9 after repeated administration, but did not significantly change the enzymatic activities of CYP3A4 and CYP2C19.

Apatinib combined with capecitabine in the treatment of advanced triple-negative breast cancer as third-line therapy: An observational study

WANG Aijuan, GUO Yiyun, WANG Zhiqiu

2020, 25(4):  433-439.  doi:10.12092/j.issn.1009-2501.2020.04.013

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AIM: To investigate the efficacy and safety of apatinib combined with capecitabine in the treatment of advanced triple-negative breast cancer (TNBC) as third-line therapy. METHODS: Sixty advanced TNBC patients, who have failed to receive second-line palliative chemotherapy, were enrolled in the Department of Oncology, Anqing Hospital Affiliated to Anhui Medical University from February 2016 to September 2019. Patients were divided into observation group (n=30, received apatinib combined with capecitabine) and control group (n=30, received capecitabine) randomly. The objective response rate (ORR), disease control rate (DCR), progression-free survival (PFS), 1-year survival rate, overall survival (OS) and adverse events between the two groups were observed and compared. RESULTS:ORR was 26.67% and DCR was 86.67% in the observation group, while 6.67% and 60.00% in the control group, respectively. ORR and DCR in the observation group were better than those in the control group (P=0.038; 0.020). Meanwhile, the median PFS was 7.0 months in the observation group, while it was 5.0 months in the control group, which indicated that the observation group exhibited a higher PFS than the control group (P=0.000). The 1-year survival rate and the median OS was 55.30% and 13.0 months in the observation group respectively, while those were 46.30% and 12.0 months in the control group respectively, the OS showed no significant differences between the two groups (P=0.258). In addition, we also found that there was significant differences in the adverse reaction such as hypertension between the two groups (P=0.000), yet it was mild and tolerable after symptomatic treatment. CONCLUSION:Apatinib combined with capecitabine in the advanced TNBC maintenance treatment has a certain survival benefit for those who failed in second-line therapy, and adverse reactions are tolerable and controllable.

Effects of SSRI antidepressant sertraline on ocular surface function in patients with postpartum depression

GE Wei, SHENG Wenyan, XU Qibing, LI Qiushi, SU Lingya, WU Shuangqing

2020, 25(4):  440-446.  doi:10.12092/j.issn.1009-2501.2020.04.014

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AIM: To investigate the effect of SSRI antidepressant sertraline on ocular surface function in patients with postpartum depression.METHODS: This was a prospective case-control study. Forty-three patients diagnosed as postpartum depression receiving sertraline for the first time were investigated with the HAMD, OSDI, TBUT, SIT, FL, tear osmolarity and IL-6, MMP-9 before medication and 2 months after medication. The changes of dry eye indexes before and after medication were compared.RESULTS:After 2 months of medication, HAMD score was significantly decreased, with statistically significant differences (P<0.01). There was no correlation between efficacy of medication and dry eye (t=0.119, P>0.05). After 2 months of medication, OSDI score increased, TBUT time shortened significantly, tear secretion decreased, FL score increased, tear osmolarity increased. The differences were statistically significant (P<0.01, P<0.01, P<0.05, P<0.01, P<0.01, P<0.01, P<0.01, respectively). The expression level of IL-6 and MMP-9 increased significantly 2 months after medication, with statistically significant differences (P<0.01 and P<0.01, respectively). There was a positive correlation between tear osmolarity and IL-6 (r=0.472, P<0.01), a positive correlation between tear osmolarity and MMP-9 (r=0.403, P<0.01), a negative correlation between tear osmolarity and BUT (r=－0.536, P<0.01), and no correlation between tear osmolarity and FL (r=0.143, P>0.05), which indicated that tear osmolarity, tear inflammatory factor and tear metalloproteinase expression level increased when dry eye occurred. CONCLUSION:Sertraline can effectively relieve and cure postpartum depression, but long-term administration can cause damage to ocular surface function, leading to an increase in the incidence of dry eye.

Temperature selection of intravesical sodium hyaluronate in the treatment of interstitial cystitis

CAI Jiangyi, LIU Yao, LI Jiacheng

2020, 25(4):  447-454.  doi:10.12092/j.issn.1009-2501.2020.04.015

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AIM: To investigate the efficacy of bladder infusion of sodium hyaluronate (HA) in the treatment of interstitial cystitis (IC) at different temperatures.  METHODS: Forty patients with IC admitted in our hospital from March 2018 to April 2019 were simply randomized and divided into two groups for prospective study, with 20 patients in each group. The temperature of perfusion fluid in normal temperature group was 37 ℃, and the temperature of perfusion fluid in high temperature group was 43 ℃. Compared the two groups before treatment, one month after treatment, and three months after treatment of interstitial cystitis problem score (ICPI), interstitial bladder symptoms score (ICSI), pre-veeing pain visual analogue score (VAS), and anxiety self-assessment scale (SAS), depression self-assessment scale (SDS), quality of life score (QOL), daily urination times, maximum bladder volume, bladder mast cell count, and immune factors ［interleukin (IL)-6, IL-10］ expression. RESULTS:The ICPI, ICSI, VAS, SAS, and SDS scores of the hyperthermia group were lower than those of the normal temperature group after one month and three months of treatment, and the QOL scores were higher than those of the normal temperature group after three months of treatment (P<0.05). After one month and three months of treatment in the hyperthermia group, the daily urination frequency was lower than that in the normal temperature group, and the maximum bladder capacity after three months of treatment was greater than the normal temperature group (P<0.05). After three months of treatment in the hyperthermia group, the mast cell counts in the bladder mucosa were lower than those in the normal temperature group (P<0.05). IL-6 and IL-10 in the bladder mucosa were lower than those in the normal temperature group after three months of treatment in the high temperature group (P<0.05). There was no significant difference in the incidence of adverse reactions (0, 5.00%) between the two groups (P>0.05). CONCLUSION:Compared with 37 ℃, intravesical instillation of HA in the treatment of IC can effectively relieve the clinical symptoms and signs of patients when the perfusion fluid is at a temperature of 43 ℃, and it improves the maximum bladder capacity and anxiety and depression, improves the quality of life of patients, which is safe and reliable. The mechanism may be related to the regulation of large cell, IL-6, IL-10 of bladder mucosa.

Effects of atorvastatin and folic acid tablets on homocysteine and carotid atherosclerosis in patients with type H hypertension

XIE Xiangmei, HUANG Zhihong, LIU Xiaoju, XUE Bing, LAN Baozhu

2020, 25(4):  455-459.  doi:10.12092/j.issn.1009-2501.2020.04.016

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AIM: To analyze the effects of atorvastatin and folic acid tablets on homocysteine and carotid atherosclerosis in patients with type H hypertension.  METHODS: A total of 140 patients with H-type hypertension admitted to our hospital from January 2016 to October 2018 were enrolled in the study. Random number table method was divided into observation group (70 cases) and control group (70 cases). The control group was treated with folic acid tablets on the basis of routine blood pressure reduction. The observation group was given atorvastatin calcium tablets on the basis of the control group, and the treatment was 10 weeks. Comparing the two groups of homocysteine, low-density lipoprotein, brachial-ankle pulse wave velocity, carotid intima-media thicknessbefore and after treatment, blood pressure and heart rate levels, and observed the occurrence of adverse reactions. During a follow-up of one year, the incidence of cardiovascular and cerebrovascular events was compared between the two groups. RESULTS:The plasma Hcy level and LDL level in the observation group was significantly lower than that in the control group (P<0.05). There was significant difference in BaPWV and IMT between the two groups (P<0.05). After treatment, the systolic and diastolic blood pressures of the two groups were treated. The systolic blood pressure and diastolic blood pressure were significantly lower than those in the control group (P<0.05), but there was no significant difference in heart rate between the two groups after treatment (P>0.05). There was no significant difference in the incidence of adverse reactions between the two groups (P>0.05). CONCLUSION:Atorvastatin combined with folic acid tablets can effectively reduce carotid atherosclerosis in patients with type H hypertension, and it can reduce the levels of Hcy and LDL , and it is clinically safe.

Effects of pharmaceutical excipients on CYP3A activity and its guiding effects on quality and efficacy consistency evaluation of generic drugs

陈思雨，陈露露，王欣桐，李晓晖，余鹏，欧阳冬生

2020, 25(4):  460-466.  doi:10.12092/j.issn.1009-2501.2020.04.017

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The prescription and technology of pharmaceutical preparations are the basis for ensuring the quality and efficacy of medicines. Pharmaceutical excipients are important part of pharmaceutical preparations. As it's known to all that some pharmaceutical excipients can affect the activity of CYP3A, and then may influence the metabolism and bioavailability of its substrates in vivo. Relative bioavailability is a key element of generic drug research in China and the United States. Our country is advancing the quality and efficacy consistency evaluation of generic drugs. Therefore, it is of great practical significance to understand the impact of pharmaceutical excipients on CYP3A and its guiding role in consistency evaluation.

Pharmacological targets and clinical application of traditional Chinese medicine and active components in the prevention and treatment for depression

FENG Rui, ZHANG Jiali, HE Mingchao, LI Yue, ZHANG Yan

2020, 25(4):  467-474.  doi:10.12092/j.issn.1009-2501.2020.04.018

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The pathological mechanism for depression is complicated due to the involvement of types of transcriptional factors and signaling pathways in the occurrence and development of depression. Traditional Chinese medicine (TCM) characterized with multi-components and multi-targets exert unique advantage in the prevention and treatment of depression. This review summarized the pharmacological action of TCM and active components on transcriptional factors (CREB, NF-κB, Nrf2) and signaling pathways (BDNF-TrkB, MAPK, GSK-3β, TLR-4-NLRP3-IL-1β) as well as elucidated the clinical application of TCM formula in the prevention and treatment for depression based on relevant literatures from home and abroad.

Progress in drug therapy for advanced triple-negative breast cancer

ZHAO Quanming, YANG Yang

2020, 25(4):  475-480.  doi:10.12092/j.issn.1009-2501.2020.04.019

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Triple-negative breast cancer is estrogen receptor, progesterone receptor and human epidermal growth factor receptor-2 negative breast cancer. The lack of specific treatment targets, there are few effective therapy for advanced triple- negative breast cancer. This article will review the current progress in drug therapy for advanced triple-negative breast cancer, including chemotherapy, targeted therapy, endocrine therapy and immunotherapy, in order to provide reference for clinical treatment.