Table of Content

Volume 24 Issue 6
26 June 2019

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Network pharmacology-based study on the mechanism of Salvia miltiorrhiza in treatment of Parkinson's disease

ZHANG Shuijing, DU Zhongyan

2019, 24(6):  601-607.  doi:10.12092/j.issn.1009-2501.2019.06.001

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AIM: To predict and screen the active constituents of Parkinson's disease and its potential targets by using network pharmacology techniques, and to explore the mechanism of prevention and treatment of Salvia miltiorrhiza on multi-component group-multi-target group-disease of Parkinson's disease. METHODS: BATMAN-TCM was used to predict and screen the active components and target of Salvia miltiorrhiza. The components and target network of Salvia miltiorrhiza in Parkinson's disease were constructed by using Cytoscape software. The PPI network was constructed by STRING database, and the target GO was analyzed by KOBAS database. Enrichment and KEGG signaling pathways. RESULTS:The 124 active components of Salvia miltiorrhiza were screened and 11 targets related to Parkinson's disease were predicted, which were mainly related to ACHE, ADORA2A, AGTR1 and other targets. Ten of them had interactions, and were screened at P<0.05. A total of 1 112 GO biological processes were obtained, the functions of which involved processes such as dopamine metabolism, catechol-containing compound metabolism. 34 signal pathways were screened, and the first 20 KEGG pathways were related to dopaminergic synapses and neuroactive ligand-receptor interactions. CONCLUSION:The mechanism of multi-component-multi-target-multi-pathway treatment of Parkinson's disease from dopamine metabolism process and neuroactive ligand-receptor interaction is revealed, which provides theoretical support for targeted drug development and clarification mechanism.

Psychophysiological observation of propofol addiction in rats

XU Wenting, YANG Liqin, ZHANG Zhongnan, LIANG Jun, JIANG Nan, JIANG Miao, GUO Wenjun, WANG Mengya

2019, 24(6):  608-614.  doi:10.12092/j.issn.1009-2501.2019.06.002

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AIM: To observe the addiction action of propofol on rats and explore the possible psychophysiological characteristics of the model rats. METHODS: Thirty-three adult male SD rats were divided into two groups randomly, i.e. Propofol Group (n=18) and Control Group (n=15). After a week of adaptive feeding, an 11-day conditioned place preference (CPP) experiment, including pretest stage, training stage and posttest stage was conducted in all rats.After CCP experiment, the electric activities from addiction-related nuclei accumbens nucleus core (Acbc) and basolateral amygdala (BLA) were extracellularly recorded, and physiological functions of electrocardiogram (ECG), electromyogram (EMG) of respiratory muscles and body temperature were simultaneously observed. The changes of electrical activities in Acbc and BLA, and physiological functions were observed after intraperitoneal injection of NS and propofol. RESULTS:(1) There was an increase of the time in the drug box for the propofol group rats (P<0.01), and an increased difference between posttest and pretest in comparison with control group rats (P<0.05). (2) The firing rate in Acbc and BLA in propofol group rats (n=10) was significantly higher than that in the control group rats (n=5) (Acbc: P<0.01, BLA: P<0.05). (3) An increased Acbc/BLA ratio of firing rate in propofol group rats was seen in comparison with control group rats (P<0.05). (4) No significant changes of firing rate were observed after injecting propofol but the respiratory rate and body temperature in propofol group rats decreased significantly (P<0.05, n=8). CONCLUSION: These results indicate that the short-term repeated administration of small dose propofol may lead to addiction, which is possibly related to the changes in reward-punishment system balance in the brain of rats.

Improvement of CGRP on vascular endothelial cells insulin resistance via activation of cAMP/PPARγ/eNOS pathway

QUAN Haiyan, LIU Yuhuan, YANG Li, YANG Fang, QIN Xuping

2019, 24(6):  615-622.  doi:10.12092/j.issn.1009-2501.2019.06.003

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AIM: To explore the protective effect and the mechanism of calcitonin gene-relatedpeptide (CGRP) on the insulin-resisted human umbilical vein endothelial cells (irHUVECs). METHODS: The irHUVECs was established using human umbilical veinendothelial cells (HUVECs), gucose oxidase (GOP-POD) method and anthrone-sulfuric acid method were employed to detect glucose consumption ability and glycogen synthesis ability, respectively. RT-PCR and Western blot were used to detect the mRNA and protein expressions of peroxisome proliferator-activated receptor gamma (PPARγ) and endothelial nitric oxide synthase (eNOS), respectively. RESULTS:The irHUVECs was successfully established when the HUVECs were treated with 33.3 mmol/L high glucose and 5 μmol/L insulin. Compared to the normal group of endothelial cells, the features of irHUVECs were a larger volume, fuzzy boundary and abnormal morphology, when cells induced were incubated with high glucose and high insulin in 24 h and 48 h, respectively, the glucose consumption was reduced by 20% and 31%, and the glycogen synthesis was reduced by 55% and 64%, meanwhile, the expressions of PPARγ and eNOS were decreased. Treatment of CGRP significantly increased about 20% glucose consumption and about 70% glycogen synthesis, and increased the expressions of PPARγ and eNOS, SQ22536 (cAMP inhibitor) can decreased the expressions of PPARγ and eNOS. CONCLUSION: CGRP could improve the endothelial cell insulin resistance, the mechanism may be related to up-regulated cAMP, and the increased expressions of PPARγ and eNOS.

Expression of miR-222 and MBD2 in gastric cancer patients and their effects on platinum chemosensitivity

DAI Lei, LUO Linghe, WU Liyan, WANG Gang, FEI Baoying

2019, 24(6):  623-629.  doi:10.12092/j.issn.1009-2501.2019.06.004

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AIM: To study the expression level of miRNA-222 and methylated CpG binding structural protein 2 (MBD2) in gastric cancer and its effect on proliferation and apoptosis of human gastric cancer cells. METHODS:Seventy-five patients with primary gastric cancer who were admitted to our hospital from February 2016 to May 2017 were selected as subjects. The expression levels of miR-222 and MBD2 in cancer tissues and adjacent tissues were detected. The correlation between miR-222 and MBD2 levels and clinicopathological parameters of gastric cancer were analyzed. The miR-222 inhibitor was transfected into gastric cancer SGC-7901 cells. The expression levels of miR-222 and MBD2 were detected by RT-PCR and Western blot, respectively. MTT assay and flow cytometry were used to detect. The effects of transfected cells on proliferation and apoptosis. RESULTS:The positive expression rate of miR-222 in gastric cancer tissues was significantly higher than that in adjacent tissues (P<0.05). The relative expression of miR-222 in gastric cancer tissues was significantly higher than that in adjacent tissues (P<0.05). The positive expression rate of MBD2 in gastric cancer tissues was significantly lower than that in adjacent tissues (P<0.05). The expression levels of miR-222 and MBD2 were significantly correlated with the degree of differentiation and TNM stage (P<0.05). The expression level of miR-222 in SGC-7901 cells transfected with miR-222 inhibitor was significantly lower than that in control group and NC group (P<0.05). The expression levels of MBD2 mRNA and protein in the cells transfected with miR-222 inhibitor were significantly higher than those in control group and NC group (P<0.05). The proliferation rate of cells transfected with miR-222 inhibitor was significantly lower than that of Control group and NC group (P<0.05), and the cell proliferation rate of miR-222 inhibitor+CDDP group was significantly lower than that of miR-222 inhibitor group and CDDP group. The apoptosis rate was significantly higher than that of miR-222 inhibitor group and CDDP group (P<0.05), and combined with CDDP could inhibit the proliferation rate of tumor cells and promote cell apoptosis. CONCLUSION: The expression of miR-222 in gastric cancer tissues is up-regulated in advanced gastric cancer tissues, and MBD2 is down-regulated significantly. The expression level of miR-222 is significantly correlated with the differentiation degree of gastric cancer and TNM stage. miR-222 can enhance the chemosensitivity of CDDP to gastric cancer，which may play a role in inhibiting the target gene MBD2 in gastric cancer.

Effect of hypoxia-inducible factor 2α on chemoresistance of hepatocellular carcinoma

YANG Shucai，ZHANG Li，LIU Liping，DENG Weijie，ZHOU Jie，LIU Hui，ZHANG Baohu，JIN Tao

2019, 24(6):  630-636.  doi:10.12092/j.issn.1009-2501.2019.06.005

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AIM: To study the effect of hypoxia-inducible factor 2α (HIF-2α) on chemoresistance of hepatocellular carcinoma and its mechanism. METHODS: Six hepatocellular carcinoma cell lines were selected and the expression of HIF-2α were detected by Western blot. The HIF-2α lentiviral particles were produced and infected with HepG2 and PLC/PRF/5 cells to prepare a cell line stably expressing HIF-2α, and Western blot was used to verify its expression. Six conventional chemotherapy drugs (fluorouracil, cyclophosphamide, epirubicin hydrochloride, mitomycin, methotrexate, and sorafenib) in five concentrations were added to HepG2-HIF2α and Control cells. MTT assay was used to detect cell inhibition rate. To study the mechanism, Western blot was used to detect the expression of drug resistance genes MDR1, LRP and MRP1 in HIF-2α overexpressing HCC cells and the corresponding control cells. RESULTS:Western blot result showed that HIF-2α was expressed in 6 hepatocellular carcinoma cell lines. The ectopic expression of HIF-2α in lentivirus-infected HepG2 and PLC/PRF/5 cells were validated by Western blot. MTT results showed that the inhibitory rate of cyclophosphamide, methotrexate and sorafenib on HepG2-HIF-2α was significantly lower than control cells (P<0.05), suggesting that the expression of HIF-2α caused chemotherapy resistance. However, the inhibition rate of fluorouracil, epirubicin hydrochloride and mitomycin on HepG2-HIF-2α cells was not different from that of the control group (P>0.05). Western blot analysis further analyzed the cause of HIF-2α on the increase of chemotherapy resistance in hepatocellular carcinoma cells. The results showed that the expression levels of drug resistance-related genes MDR1, LRP and MRP1 were higher in HepG2-HIF-2α cells and PLC/PRF/5-HIF-2α cells than those in the control group (P<0.05). CONCLUSION: In hepatocellular carcinoma, HIF-2α can increase the chemotherapy resistance of hepatocellular carcinoma cells by up-regulating the expression of MDR1, LRP and MRP1 resistance genes.

α-synuclein activates NLRP3 inflammatory bodies to mediated nerve cell pyroptosis

HAN Chenyang, ZHANG Xiaoling, YANG Yi, GUO Li, GUAN Qiaobing

2019, 24(6):  637-643.  doi:10.12092/j.issn.1009-2501.2019.06.006

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AIM: To study the effect and mechanism of α-synuclein on nerve cell pyroptosis.  METHODS: HT22 cell was cultured in vitro and interfered by α-synuclein. Cell viability was measured by CCK-8 method and IC50 value was determined. After setting up the control group (Con group), TUNEL staining was used to detect the pyroptosis level of cells, immunofluorescence was used to observe the expression of GSDMD-N, Western blot was used to detect the expression levels of NLRP3, ASC, GSDMD, GSDMD-N and Caspase-1 in cells, and enzyme-linked immunosorbent assay was used to detect the expression levels of IL-18 and IL-1β in culture medium. After HT22 was pretreated with Caspase-1 inhibitor, the cells were also treated with α-synuclein of IC50 value, and the α-synuclein group (only α-synuclein intervention) and α-synuclein+ML132 group were set up to detect the level of pyroptosis. RESULTS:The IC50 value of α-synuclein was 50 nmol/L. After the intervention of α-synuclein, the number of nerve cells increased significantly. TUNEL staining showed that the number of positive cells was more than that of Con group. Immunofluorescence staining showed that the expression level of GSDMD-N in cells was up-regulated. The expression levels of NLRP3, ASC, GSDMD-N and Caspase-1 in cells were significantly higher than those in Con group, while the level of GSDMD was down-regulated in cell culture medium. The levels of IL-18 and IL-1β were up-regulated. After treatment with Caspase-1 inhibitor, the pyroptotic level of 50 nmol/L α-synuclein was significantly lower than that of α-synuclein group. Immunofluorescence staining showed that the level of GSDMD-N was down-regulated. The expression levels of NLRP3, ASC, GSDMD-N and Caspase-1 in cells were significantly lower than those in α-synuclein group, while the level of GSDMD was up-regulated. The levels of IL-18 and IL-1β in culture medium were down regulation. CONCLUSION:α-synuclein may play a role in the development of Parkinson's disease by activating NLRP3 inflammatory bodies.

Expression and mechanism of p38MAPK signaling pathway in chronic pulpitis rats with pain

LI Min, WANG Shanshan, LIU Zhonghe, JIANG Deguo

2019, 24(6):  644-649.  doi:10.12092/j.issn.1009-2501.2019.06.007

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AIM: To investigate the expression of p38MAPK signaling pathway in chronic pulpitis pain rats and its mechanism of action.  METHODS: Twenty-five Wistar male rats of SPF grade were selected and divided into 3 groups, 5 in normal group, 15 in model group and 5 in MAPK inhibitor group. In the MAPK inhibitor group and the model group, the model of pulpitis was prepared. After successful modeling, the MAPK inhibitor group was injected with SB203580 at a dose of 5 mg/kg from the jugular vein for 14 days. The MAPK inhibitor group was selected 14 days after modeling. Five rats in the model group were sacrificed at 3, 7 and 14 d after modeling, respectively. Their pulp tissue pathology, serum interleukin-6 (IL-6), IL-8 content, p-p38MAPK, ERK, p-ERK, p38MAPK protein expression and p38MAPK, ERK mRNA expression were observed.RESULTS:Compared with the normal group, the length of apical damage, the width of the periapical periodontal membrane and the root necrosis rate of the first molar were significantly increased at 3, 7 and 14 days after modeling; while those in the MAPK inhibitor group were significantly lower (P<0.05). The levels of serum IL-6 and IL-8 in rats at 3, 7 and 14 days after modeling were significantly increased. Compared with 3, 7 and 14 days after modeling, serum IL-6 and IL-8 levels in rats with MAPK inhibitor group were significantly lower (P<0.05). Compared with the normal group, the expression of p-p38MAPK and p-ERK protein in the pulp tissue of the model group was increased. Compared with the model group, the MAPK inhibitor group The expression of p-p38MAPK and p-ERK protein in rat dental pulp tissue decreased (P<0.05).CONCLUSION: Activation of p38MAPK signaling pathway in dental pulp tissue of rats with chronic pulpitis increases the degree of inflammatory response, promotes disease progression, and inhibits p38MAPK signaling pathway, which can effectively reduce the expression of inflammatory factors.

Evaluation of Poly NP test and MPNP test for rapid detection of the polymyxin resistance Klebsiella pneumoniae

LIU Yanling, ZENG Lingbing, HU Niya, CHEN Kaisen, HU Xuefei, HU Longhua

2019, 24(6):  650-657.  doi:10.12092/j.issn.1009-2501.2019.06.008

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AIM: To evaluate the screening capacity of rapid polymyxin NP test (Poly NP test) for the detection of polymyxin B resistance Klebsiella pneumoniae (PolR-KP) and the modified rapid polymyxin Nordmann/Poirel test (MPNP) for the MCR-producing in K.pneumoniae. METHODS: A total of 48 isolates were selected, in which, 14 strains were polymyxin sensitive Klebsiella pneumoniae (PolS-KP), 26 strains were PolR-KP, 3 strains were polymyxin intrinsic resistance and 5 strains of nonfermenters. The rapid polymyxin NP test was used to detect the polymyxin resistance phenotype and the results were compared with the MICs determined by the BMD method, which was taken as the gold standard. The PCR and sequencing were used to screen the polymyxin resistance determinants caused by chromosomal mutations-the two-component phoP/phoQ, pmrA/pmrB and the mgrB gene; and also screen the presence of mcr-1, mcr-2, mcr-3, mcr-4, mcr-5, mcr-6, mcr-7 and mcr-8. The MCR-producing Klebsiella pneumoniae isolates were detected by MPNP test. RESULTS:All the polymyxin intrinsic resistance strains were positive in Poly NP test, of the 26 isolates PolR-KP, 24 PolR-KP strains were positive in Poly NP test and only 2 strains showed negative result. The sensitivity and specificity of Poly NP test were 93.1% and 100%. Of the 22 isolates which polymyxin resistance was associated with chromosome-encoded mechanisms, 12 isolates had mutations in the pmrA/pmrB two-component system, 4 isolates in the phoP/phoQ two-component system and 6 isolates had different alterations in mgrB gene. 4 strains of MCR-producing K. pneumoniae were detected in PolR-KP and the MPNP test showed lack of inhibitory effect of EDTA on the mcr-positive K. pneumoniae strains. CONCLUSION: The Poly NP test is an easy-to-perform, rapid, sensitive, and specific test and making it a potential useful clinical technique for the detection of PolR-KP, while the MCR-positive K.pneumoniae strains are not identified by the MPNP test. Further methods suitable for rapid detection of the MCR-producing in K.pneumoniae are uegently needed.

Inhibitory effect and mechanism of quercetin on secondary endotoxemia in rats with Neisseria infection

LI Zhengzheng, CHEN Jin, PAN Zhenzhen, PAN Sipei, ZHANG Nan

2019, 24(6):  658-664.  doi:10.12092/j.issn.1009-2501.2019.06.009

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AIM: To analyze the effect of quercetin on the inhibition of secondary endotoxemia in rats with Neisseria meningitis. METHODS: Sixty Wistar male rats of SPF grade were randomly divided into six groups: model group, normal group, positive control group, low dose quercetin group, medium dose quercetin group and high dose sputum. 10 rats in each group, except for the normal group, the other rats were prepared with Neisseria infection meningitis model. After the model was established, the positive control group was intragastrically administered with a 2 mL dose of 100 mg/L dexamethasone solution. Low-dose, medium-dose and high-dose quercetin groups were intragastrically administered with 2 mL doses of 50, 100, 200 mg/L quercetin solution, and normal group and model group rats were given equal doses of normal saline. Rats were administered at intervals of 12 h for a total of five doses. The survival rate of rats in each group was observed at 12, 24, 36, 48 and 60 hours. The body temperature of rats was measured once every 12 hours. Serum IL-6, TNF-α, lipopolysaccharide, IL-1β and IL-8 content were detected by ELISA. Western blot was used to detect the p65 and IκBα protein expression, RT-PCR was used to detect the relative expression of brain tissue Toll-like receptor 4 (TLR4) mRNA and CD68 mRNA. RESULTS:All the rats in the model group died within 24 hours after modeling, and the survival rate of the model group was lower than that of the normal group. The survival rate of the medium dose and high dose quercetin group was better than that of the model group, the positive control group and the low dose quercetin group. The difference was statistically significant (P<0.05). The serum levels of IL-6, TNF-α, IL-1β and IL-8 in the model group were significantly higher than those in the normal group (P<0.05). The levels of IL-6, TNF-α, IL-1β and IL-8 in the quercetin groups were significantly lower than those in the model group. Among them, the rats in the middle dose and high dose quercetin groups decreased significantly, and the differences between the model group and the positive control group were statistically significant (P<0.05). The expression of IκBα protein in the model group was lower than that in the normal group, and the expression of p65 protein was higher than that in the normal group. The expression of IκBα protein in the middle dose and high dose quercetin group was higher than that in the model group and the positive control group. The p65 protein in the middle dose and high dose quercetin group was lower than the model group and the positive control group, and the difference was statistically significant (P<0.05). CONCLUSION: Quercetin can inhibit the NF-κB inflammatory pathway in rats with Neisseria meningitis, reduce the content of serum inflammatory factors, and improve the survival rate of rats.

Effect of astragaloside on the apoptosis of human renal carcinoma cell (ACHN) via SDF-1/CXCR4 signaling pathway

ZOU Zhaoyin, ZHENG Fu, XU Xianshun, DING Wenjuan, CHEN Yan, PU Daojing

2019, 24(6):  665-669.  doi:10.12092/j.issn.1009-2501.2019.06.010

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AIM: To observe the effect of astragaloside on the proliferation, apoptosis of human renal carcinoma cell (ACHN) and the activation of SDF-1/CXCR4 signaling pathway.  METHODS: ACHN cells were co-cultured with 20, 60 and 120 μmol/L of astragaloside. CCK-8 assay was used to detect the effect of astragaloside on proliferation inhibitory rates of ACHN cells. The apoptotic rates of ACHN cells were analyzed using flow cytometer. Western Blot was used to determine the expression levels of Csapase-8, Bax, Bcl-2, SDF-1 and CXCR4 proteins in ACHN cells. RESULTS:After treatment with 20, 60 and 120 μmol/L of astragaloside, the proliferation inhibitory rates of ACHN cells were significantly increased with time- and dose-dependent manner (P<0.05). The apoptotic rates of ACHN cells in astragaloside groups were significantly increased compared with control group (P<0.05). The expression levels of Csapase-8 and Bax in astragaloside groups were significantly increased, and the expression levels of Bcl-2, SDF-1 and CXCR4 were significantly decreased compared with control group (P<0.05). CONCLUSION: Astragaloside can inhibit the proliferation of ACHN cells and induce the apoptosis of ACHN cells, and its mechanism was associated with the inhibition of SDF-1/CXCR4 cascade.

Classification and application of logic check in clinical trail

ZHOU Bei, YU Hao

2019, 24(6):  670-674.  doi:10.12092/j.issn.1009-2501.2019.06.011

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Clinical trial data cleaning is very important in the process of clinical trials. Only integrated, cleaned data can be used for statistical analysis. The purpose of this paper is to discuss the compose of Data Verification Plan and the classification of logical verification in the process of data cleaning, and in further to discuss the possible influence on data collection and cleaning by the trend of clinical trial digitization in recent years.

Bioequivalence of bosentan tablets in healthy Chinese volunteers

XU Yichao, LOU Honggang, RUAN Zourong, CHEN Jinliang, JIANG Bo, SHAO Rong, YANG Dandan

2019, 24(6):  675-680.  doi:10.12092/j.issn.1009-2501.2019.06.012

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AIM: To evaluate the bioequivalence of a single oral dose of bosentan tablets of test and reference formulations in healthy Chinese volunteers. METHODS: It was a randomized, open-label, two sequences, two period crossover study. A single oral dose of 125 mg test and reference formulations were given to 24 healthy Chinese volunteers under fasting and fed conditions. The concentrations of bosentan and its active metabolite hydroxybosentan in plasma were determined by high performance liquid chromatography tandem mass spectrometry. The pharmacokinetic parameters were calculated and the bioequivalence was compared by non-compartment model of WinNonlin program. RESULTS:The bosentan's major pharmacokinetic parameters of test and reference under fasting condition were as follow: Cmax were (2.72±1.09) and (2.59±1.02) μg/mL, AUC0-t were (13.51±4.55) and (12.61±4.25) μg·mL-1·h, AUC0-∞ were (14.04±4.46) and (13.02±4.24) μg·mL-1·h, tmax were (3.75±0.81) and (3.98±0.76) h, t1/2 were (2.70±0.88) and (2.77±0.91) h。The bosentan's major pharmacokinetic parameters of test and reference under fed condition were as follow: Cmax were (2.84±0.68) and (2.95±0.99) μg/mL, AUC0-t were (14.66±4.27) and (15.34±6.09) μg·mL-1·h, AUC0-∞ were (15.04±4.41) and (15.68±6.19) μg·mL-1·h, tmax were (3.98±1.05) and (3.98±1.21) h, t1/2 were (2.55±0.65) and (2.71±0.63) h. There was no significant difference among the order of administration, formulations and cycles (P>0.05); The 90% confidence intervals of AUC0-t, AUC0-∞ and Cmax were in the range of 80%-125% by double-sided t test at the level of α=0.05.CONCLUSION: The two formulations of bosentan tablets were bioequivalent in Chinese healthy volunteers under fasting and fed condition.

Investigation of curative effect and mechanism of Chaihu Jiedu Decoction on condyloma acuminatum

BU Zhangyu, WU Liming, YU Xiaohong, LIU Jue, DENG Lin

2019, 24(6):  681-686.  doi:10.12092/j.issn.1009-2501.2019.06.013

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AIM: To observe the clinical efficacy of the Chaihu Jiedu Decoction in the treatment of condyloma acuminatum (CA), and to explore the possible effective mechanism. METHODS: A total of 120 patients with CA (age range from 16 to 65) who were admitted and treated in our hospital from June 2015 to September 2017 were selected. All patients were randomly divided into three groups: group A received Chaihu Jiedu Decoction（n=40）, group B received α-2b recombinant human interferon（n=39, one of the patients withdrew from the study due to adverse reactions after 2 weeks of medication）and group C received no systemic therapy（n=40）. The clinical efficacy, recurrence rate, recurrence frequency and adverse drug adverse reaction of the three groups were compared. Peripheral blood T lymphocyte subsets and NK cells before and after the treatment of CA patients in group A were detected by flow cytometry, and IL-2, IL-4, IL-10, IL-12, IFN-γ in peripheral venous blood serum were detected by ELISA. RESULTS:Except for the one withdrawal case from group B, after 12 weeks follow-up, the rate and the average times of recurrence in all of the CA cases were 51.26%（61/119）and 1.6, respectively. The rate and the average times of recurrence in group A and B were lower than those of group C（P<0.01）, but the differences between group A and B were not statistically significant（P>0.05）. At the end of one month treatment with Chaihu Jiedu Decoction, the CD4+T cell percentage, the CD4+/CD8+ cell ratio, and the NK cell percentage were all increased（P<0.01）in peripheral blood of CA patients, but the differences of CD3+T cell percentage and CD3+T cell percentage were not statistically significant. Also, at the end of treatment with Chaihu Jiedu Decoction, the peripheral blood serum IL-2 of CA patients increased（P<0.01）, the IL-12（P<0.01）, IFN-γ（P<0.01）and IL-10 decreased（P<0.05）, while the difference of IL-4 content was not statistically significant（P>0.05）. CONCLUSION:The Chaihu Jiedu Decoction has significant clinical effect on CA. The efficacy is comparable to intramuscular injection of α-2b recombinant human interferon. The effective mechanism may be the improvement of cell-mediated immunity function by promoting the number and function of CD4+T cell, and the increase of the number and function of NK cells by promoting the increase of cytokines Il-2 and IFN-γ secreted by CD4+T cells.

Effect of magnesium sulfate mixed ropivacaine femoral nerve block on lower extremity tourniquet reaction in the patients with knee ligament repair

WANG Hongzhu, LIN Xianju, ZHU Lijun

2019, 24(6):  687-692.  doi:10.12092/j.issn.1009-2501.2019.06.014

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AIM: To study the effect of magnesium sulfate mixed ropivacaine femoral nerve block on lower extremity tourniquet reaction in the patients with knee ligament repair. METHODS:A total of 83 patients of underwent arthroscopic knee ligament repair who received therapy from February 2016 to February 2018 in our hospital were selected as research objects, they were randomly divided into treatment group (n=42) and control group (n=41), the two groups were operated under the guidance of ultrasound machine, the treatment group was given 0.375% ropivacaine 15 mL (containing Magnesium Sulfate 0.75 g) mixed injection, and the control group was given 0.375% ropivacaine 15 mL injection, after successful anesthesia, the tourniquet was attached to the root of the thigh and then inflated. The onset time, duration and analgesic time of sensory and motor block, the changes of the hemodynamics, serum malondialdehyde (MDA) and tumor necrosis factor-alpha (TNF-α) at different time points were compared between the two groups, and the incidence of tourniquet reaction and adverse drug reactions were recorded. RESULTS:Compared with the control group, the onset time of sensory block and motor block in the treatment group was shorten, and the duration and analgesic time were extend (P<0.05); there were significant differences in systolic blood pressure (SBP), diastolic blood pressure (DBP) and heart rate (HR) between the two groups at different time points(P<0.05); the serum MDA and TNF-α at T7 and T8 in the treatment group were higher than those in the control group(P<0.05); the tourniquet reactions were 7.14%（3/42）and 24.39%（10/41） respectively in the two groups, the difference was statistically significant(P<0.05); there was no significant difference in the incidence of adverse reactions between the two groups（P>0.05). CONCLUSION:Magnesium sulfate mixed ropivacaine femoral nerve block is well for lower extremity surgery，which can effectively maintain hemodynamic stability, prolong block effect, relieve oxidative stress and inflammatory reaction, reduce tourniquet reaction, and has high safety. It is worthy of application and promotion.

Therapeutic effect and related factors of human fibrinogen on traumatic traumatic coagulopathy

ZHU Songzhi

2019, 24(6):  693-698.  doi:10.12092/j.issn.1009-2501.2019.06.015

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AIM:To investigate the clinical efficacy of human fibrinogen(FIB) in patients with emergency traumatic coagulopathy and the related factors in patients with traumatic coagulopathy. METHODS:One hundred patients with severe trauma admitted to our hospital from June 2015 to June 2018 were included in the study. According to whether the traumatic coagulopathy，they were classified into coagulopathy group (80 cases) and non-coagulopathy group (20 cases). The acute physiology and chronic health evaluation II (APACHE II) and the injury severity score (ISS) of the first 24 h after admission，hypothermia and hypoperfusion incidence were compared between the patients in the coagulopathy group and the non-coagulopathy group. Logistic multivariate regression analysis was used to analyze the related factors of traumatic coagulopathy in patients with severe trauma. Eighty patients with traumatic coagulopathy were randomly divided into observation group (40 cases) and control group (40 cases). The control group was given conventional medical treatment. The observation group was given human fibrinogen based on the control group. The coagulation function indexes and the clinical blood volume，hospitalization time and mortality of of the observation group and the control group were compared.RESULTS:Compared with before treatment，the prothrombin time (PT)，activated partial thromboplastin time (APTT) and D-dimer levels were decreased，the level of fibrinogen (FIB) was significantly increased in the observation group and the control group after treatment(P<0.05). The red blood cell(RBC) infusion volume，fresh frozen plasma(FFP) infusion volume，intensive care unit(ICU) time and mortality rate of the observation group were significantly lower than those in the control group(P<0.05).Traumatic coagulation patients with PT>18 s (r=0.623，P=0.024)，APTT>60 s (r=0.722，P=0.018)，thrombin time(TT>15 s (r=0.719，P=0.033) were positively correlated with ISS scores. Base deficit(BD)≥6 (OR=3.678，95%CI=1.065-9.417)，Glasgow coma scale(GCS)≤8 (OR=5.299，95%CI=1.122-8.711) and platelet(PLT) count (OR=0.982，95%CI=0.971-0.996) were the independent predictors of traumatic coagulopathy in patients with severe trauma.CONCLUSION:Human FIB can effectively improve the coagulation function of patients with traumatic coagulopathy，reduce the mortality rate，shorten the ICU time，and there is certain correlation between traumatic coagulopathy and trauma severity，combined with shock，hypothermia and severe craniocerebral injury.

Effects of agomelatine on depressive disorder and brain-derived neurotrophic factor ZHOU Yancan

ZHOU Yancan

2019, 24(6):  699-703.  doi:10.12092/j.issn.1009-2501.2019.06.016

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AIM: To investigate the efficacy of agomelatine in the treatment of depressive disorder and its effect on brain-derived neurotrophic factor (BDNF). METHODS: A total of 90 patients with depression admitted to our hospital from March 20 to December 2018 were randomly divided into observation group (45 cases) and control group (45 cases). The observation group was treated with agomelatine, the control group was treated with venlafaxine, and the clinical efficacy, BDNF level and adverse reactions were observed after 4 weeks. RESULTS:The effective rate of the observation group was 73.33% and the effective rate of the control group was 62.22%. There was no significant difference in the effective rate between the two groups (P>0.05). The HAMD and HAMA scores of the two groups were significantly lower than those before treatment (P<0.05), the HAMD scores of the observation group were significantly lower than those of the control group (P<0.05), and the HAMA scores of the observation group were significantly higher than the control group (P<0.05). Pearson correlation analysis showed that BDNF level was negatively correlated with HAMD and HAMA scores (P<0.05), and positively correlated with HAMD reduction rate (P<0.05). There was no significant difference in adverse reactions between the two groups (P>0.05).CONCLUSION:Agomelatine can effectively improve the symptoms of depression and anxiety, significantly reduce the level of BDNF and has better safety.

Prognostic factors and safety study of R0 resection colorectal cancer patients received capecitabine based adjuvant chemotherapy in real world

LI Rongzhen, MEI Jiazhuan, XIA Yunzhan, JI Jie

2019, 24(6):  704-711.  doi:10.12092/j.issn.1009-2501.2019.06.017

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AIM: To investigate the prognostic factors and safety of R0 resection colorectal cancer patients received capecitabine based adjuvant chemotherapy in real world. METHODS: From Jan 2010 to December 2017, a total of 279 CRC patients underwent R0 surgical resection and received capecitabine based adjuvant chemotherapy were included in this study. Baseline characteristics data were collected through the hospital case system. The early follow-up was done during the fixed period of adjuvant chemotherapy in the hospital. Subsequent follow-up of the prognosis was telephone follow-up every three months. Disease-free survival (DFS) and overall survival (OS) were analyzed by Kaplan-Meier survival analysis. Adverse reactions were evaluated according to CTCAE 4.0. Adverse reactions above grade 2 were recorded. RESULTS:The median age of the 279 patients included in the study was 51, ranging from 19-82.167 cases (59.86%) were male. 180 patients (64.52%) were ECOG 0.173 cases of colon cancer and 106 cases of rectal cancer were included. In terms of tumor differentiation, 58 patients had well differentiation, 192 patients had intermediate differentiation, and 17 patients had poor differentiation. 58 cases of stage II patients and 221 cases of stage III patients were involved. In the state of mismatched repair gene (MMR), 19 patients with mismatched repair gene deletion (dMMR), 176 patients with mismatched repair gene presence (pMMR), and 84 patients was no detection. There were 73 patients with T1-2 and 206 patients with T3-4. 71 patients received capecitabine monotherapy and 208 patients received capecitabine combined with oxaliplatin adjuvant chemotherapy. All of the 279 CRC patients were of available for prognosis evaluation. The median disease-free survival (DFS) of the 265 CRC patients were 4.7 years (95%CI: 4.12-5.33), the median overall survival (OS) were 6.6 years (95%CI: 5.74-7.27). Adjusted in multivariate Cox regression analysis for OS, ECOG score and pathological staging were independent factors for OS. In terms of adverse reactions, neutropenia, hand and foot syndrome, diarrhea and other adverse reactions with a higher incidence of grade 2 or above were found. Overall adverse events were manageable and controllable. CONCLUSION:Capecitabine-based adjuvant chemotherapy has superior prognosis and safety in patients with colorectal cancer after R0 resection.

p73 gene and spermatogenesis and maturation

WANG Tongsheng，WU Delin，LI Li, NA Sha

2019, 24(6):  712-715.  doi:10.12092/j.issn.1009-2501.2019.06.018

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p73 is a member of the p53 family. Its structure and function are very similar to p53. It has been well known in inhibiting the occurrence and development of tumors. Recent studies have found that p73 gene regulates spermatogenesis and maturation and affects male reproductive ability. Therefore, its unique role in male reproduction has attracted much attention. The p73 gene has two different functional subtypes (TAp73 and delta Np73), which not only affect the adhesion and migration between the germ cells and the Sertoli cells, but also affect the apoptosis of the sperm. In this paper, the structure of the p73 gene and its biological function in the maturation of germ cells will be briefly described in order to provide new ideas for the diagnosis and treatment of male reproductive system diseases.

Research progress of extracellular matrix on skin wound repair

CHEN Jia, LIU Qingwu, HE Xiujuan, LI Ping

2019, 24(6):  716-720.  doi:10.12092/j.issn.1009-2501.2019.06.019

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Extracellular matrix plays an important role in the repair of skin wounds. In recent years, the research on wound healing effect and mechanism of important ECM components has been deepened. This review will expound the research progress of important ECM involved in wound repair, and provide a new idea for clinical promotion of wound repair.