Effect of LncRNA LUCAT1 targeting miR-502-5p on the biological behavior of human retinoblastoma cells / 中华实验眼科杂志

Objective:To investigate the effects of long noncoding RNA (LncRNA) lung cancer associated transcript 1 (LUCAT1) targeting microRNA (miR)-502-5p on the proliferation, migration and invasion of human retinoblastoma (RB) cells.Methods:RB tissue samples were collected from 27 RB patients who underwent eyeball enucleation in Henan Eye Hospital from May 2019 to January 2021.Another 27 normal retinal tissue specimens were collected from 12 patients with eyeball rupture, 7 with eyeball atrophy, and 8 with eyeball penetrating injury combined with pigment film incarceration who underwent the eyeball enucleation in Henan Eye Hospital during the same period.The expressions of LncRNA LUCAT1 and miR-502-5p in RB tissues, cell lines (Y-79, WERI-Rb-1, HXo-RB44) and human retinal epithelial cells (ARPE-19) were detected by real-time quantitative PCR.Y-79 RB cell was divided into control group, small interfering RNA (si)-LncRNA LUCAT1 group, si-control (con) group, pcDNA group, pcDNA-LncRNA LUCAT1 group, miR-con group, miR-502-5p group, si-LncRNA LUCAT1+ anti-miR-con group and si-LncRNA LUCAT1+ anti-miR-502-5p group, and cells in different groups were transfected with corresponding reagents.The expressions of MMP2 and MMP9 proteins were detected by Western blot.Cell proliferation activity was assayed by cell counting kit 8.Cell proliferation capability was detected by colony formation assay.Cell migration and invasion ability were determined by Transwell assay.The targeting regulation of LncRNA LUCAT1 against miR-502-5p was confirmed by dual luciferase reporter assay and real-time quantitative PCR.The study protocol was approved by the Ethics Committee of Henan Eye Hospital (No.HNEECKY-2021[32]). Written informed consent was obtained from guardians of subjects.Results:LncRNA LUCAT1 expression level in RB tissue was 2.73±0.34, which was significantly higher than 1.00±0.15 in normal retinal tissue ( t=24.190, P<0.001). The miR-502-5p expression level in RB tissues was 0.42±0.06, which was significantly lower than 1.00±0.13 in normal retinal tissue ( t=21.049, P<0.001). LncRNA LUCAT1 expression level was significantly higher and the miR-502-5p expression level was significantly lower in human RB cell lines Y-79, WERI-Rb-1 and HXO-RB44 than those in ARPE-19 cells, with statistically significant differences (all at P<0.05). The LncRNA LUCAT1 expression, the relative expressions of MMP2 and MMP9 proteins, the absorbance ( A) value, and the number of proliferated, migrating and invading Y-79 cells in si-LncRNA LUCAT1 group were significantly reduced in comparison with control group, and the differences were statistically significant (all at P<0.05). The miR-502-5p expression level was higher, and the relative expression levels of MMP2 and MMP9, A value, as well as the number of proliferated, migrating and invading Y-79 cells were lower in miR-502-5p group than in miR-con group, showing statistically significant differences ( t=20.274, 14.884, 14.181, 12.692, 17.749, 20.889, 21.913; all at P<0.001). The miR-502-5p expression level was lower and the relative expression levels of MMP2 and MMP9, A value as well as the number of proliferated, migrating and invading Y-79 cells were higher in si-LncRNA LUCAT1+ anti-miR-502-5p group than in si-LncRNA LUCAT1+ anti-miR-con group, showing statistically significant differences ( t=14.097, 15.839, 15.757, 11.860, 16.235, 16.565, 16.487; all at P<0.001). When co-transfected with LncRNA LUCAT1-wild type, the relative luciferase activity of miR-502-5p group was lower than that of miR-con group, and the difference was statistically significant ( t=16.379, P<0.001). The LncRNA LUCAT1 expression level was higher and the miR-502-5p expression level was lower in pcDNA-LncRNA LUCAT1 group than in pcDNA group, and the differences were statistically significant (both at P<0.05). The LncRNA LUCAT1 expression level was lower and the miR-502-5p expression level was higher in si-LncRNA LUCAT1 group than in si-con group, and the differences were statistically significant (both at P<0.05). Conclusions:Inhibition of LncRNA LUCAT1 can attenuate the proliferation, migration and invasion ability of human RB cells by the targeting up-regulation of miR-502-5p.

Effect of traditional Chinese medicine on portal vein thrombosis in patients with liver cirrhosis and its medication characteristics / 临床肝胆病杂志

Objective To investigate the therapeutic effect of traditional Chinese medicine (TCM) on portal vein thrombosis (PVT) in patients with liver cirrhosis and its medication characteristics. Methods A retrospective analysis was performed for 89 patients with liver cirrhosis and PVT who were hospitalized and treated in Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, and according to whether TCM treatment was applied in combination, they were divided into TCM group with 59 patients and control group with 30 patients. Related data were collected for the two groups, including demographic data, laboratory examination, radiological examination, gastroscopy, history of surgery, portal hypertension-related complications, medication, and follow-up data. The independent samples t -test was used for comparison of normally distributed continuous data between two groups, and the Mann-Whitney U rank sum test was used for comparison of non-normally distributed continuous data between two groups; the chi-square test or the Fisher's exact test was used for comparison of categorical data between two groups. An ordinal polytomous Logistic regression analysis was used for multivariate analysis. TCM Inheritance Computing Platform (V3.0) was used to perform a drug effect cluster analysis of TCM prescriptions. Results The multivariate logistic regression analysis showed that esophageal and gastric varices (odds ratio [ OR ]=3.144, 95% confidence interval [ CI ]: 1.221-8.094), PVT involving the portal vein (PV) and the superior mesenteric vein (SMV) ( OR =51.667, 95% CI : 3.536-754.859), PVT involving PV+spleen vein (SV)+SMV ( OR =13.271, 95% CI : 2.290-76.928), cavernous transformation of the portal vein ( OR =11.896, 95% CI : 1.172-120.696), and TCM intervention ( OR =0.348, 95% CI : 0.129-0.938) were influencing factors for the outcome of PVT in liver cirrhosis. Follow-up results showed that compared with the control group, the TCM group had a significantly lower progression rate (16.95% vs 56.67%, P < 0.001) and a significantly lower incidence rate of variceal rupture and bleeding (8.47% vs 33.33%, P < 0.001). Effective TCM drugs with a relatively high frequency of use included deficiency-tonifying drugs (359 times, 34.6%), blood-activating and stasis-resolving drugs (202 times, 19.5%), and diuresis-inducing and dampness-draining drugs (180 times, 17.3%); the TCM drugs with a relatively high frequency of use included Astragalus membranaceus (57 times, 8.7%), Angelica sinensis (50 times, 7.6%), and leech (48 times, 7.3%); TCM drug combinations with a relatively high frequency of use included Astragalus membranaceus+Angelica sinensis, Astragalus membranaceus+leech, Angelica sinensis+leech, and Astragalus membranaceus+Angelica sinensis+leech. Conclusion Qi-tonifying, blood-activating, and stasis-breaking drugs, such as Astragalus membranaceus, Angelica sinensis, and leech, can promote the stabilization or recanalization of PVT in liver cirrhosis and reduce the incidence rate of bleeding events due to portal hypertension.

The effect of lncRNA ADPGK-AS1 on the proliferation and apoptosis of retinoblastoma cells by targeting miR-200b-5p / 中华肿瘤杂志

Objective: To explore the effect of lncRNA ADPGK-AS1 on the proliferation and apoptosis of retinoblastoma cells and its possible mechanism. Methods: The tumor tissues of 31 patients with retinoblastoma admitted to Henan Provincial Eye Hospital from February to June 2020 and their corresponding normal tissues adjacent to the cancer were collected. The expression levels of lncRNA ADPGK-AS1 and miR-200b-5p in retinoblastoma tissues and normal adjacent tissues were detected by real-time fluorescence quantitative polymerase chain reaction (qRT-PCR). Human retinal epithelial cell ARPE-19, human retinoblastoma cell Y-79 and WERI-Rb-1 were cultured in vitro. The expression levels of lncRNA ADPGK-AS1 and miR-200b-5p were detected by qRT-PCR. Y-79 cells were randomly divided into si-con group, si-lncRNA ADPGK-AS1 group, miR con group, miR-200b-5p group, si-lncRNA ADPGK-AS1+ anti-miR con group, and si-lncRNA ADPGK-AS1+ anti-miR-200b-5p group. The proliferation, cloning and apoptosis of cells in each group were detected by tetramethylazol blue method, plate cloning test and flow cytometry, respectively. The targeting relationship between lncRNA ADPGK-AS1 and miR-200b-5p was detected by double luciferase report test, and the expression level of cleaved-caspase-3 protein was detected by western blot. Results: Compared with the adjacent tissues, the expression of lncRNA ADPGK-AS1 in retinoblastoma tissues was increased (P<0.05), while the expression of miR-200b-5p was decreased (P<0.05). Compared with ARPE-19 cells, the expression of lncRNA ADPGK-AS1 in Y-79 and WERI-Rb-1 cells was increased (P<0.05), while the expression of miR-200b-5p was decreased (P<0.05). Compared with the si-con group, the cell viability of the si-lncRNA ADPGK-AS1 group was reduced (1.06±0.09 vs 0.53±0.05, P<0.05), the number of cell clone formation was reduced (114.00±8.03 vs 57.00±4.13, P<0.05), while the apoptosis rate [(7.93±0.68)% vs (25.43±1.94)%] and the protein level of cleaved-caspase-3 were increased (P<0.05). Compared with the miR-con group, the cell viability of the miR-200b-5p group was decreased (1.05±0.08 vs 0.57±0.05, P<0.05), the number of cell clone formation was decreased (118.00±10.02 vs 64.00±5.13, P<0.05), while the apoptosis rate [(7.89±0.71)% vs (23.15±1.62)%] and the protein level of cleaved-caspase-3 were increased (P<0.05). lncRNA ADPGK-AS1 could target the expression of miR-200b-5p. Compared with the si-lncRNA ADPGK-AS1+ anti-miR-con group, cell viability of the si-lncRNA ADPGK-AS1+ anti-miR-200b-5p group was increased (0.53±0.04 vs 1.25±0.10, P<0.05), and the number of cell clones was increased (54.00±4.39 vs 125.00±10.03, P<0.05), while the rate of apoptosis [(25.38±1.53)% vs (9.76±0.71)%] and the protein level of cleaved-caspase-3 were decreased (P<0.05). Conclusion: Interfering with the expression of lncRNA ADPGK-AS1 could inhibit the proliferation and clone formation and induce apoptosis of retinoblastoma cells by targeting the expression of miR-200b-5p.

Efficacy of Fuzheng Huayu Tablets for Treating Pulmonary Inflammation in COVID-19 Patients / 中国实验方剂学杂志

ObjectiveTo observe the efficacy and safety of Fuzheng Huayu tablets （FHT） for treating pulmonary inflammation in patients with coronavirus disease 2019 （COVID-19）. MethodA total of 70（4 cases were lost to follow-up, and 66 cases were finally completed） COVID-19 patients were recruited from February 1 to April 15 in 2020. They were assigned to a control group （35 patients） and a FHT group （31 patients）. The patients in the control group received routine treatment alone and those in the FHT group received FHT in addition to routine treatment. The primary outcome was the ratio of patients showing improvement in chest computed tomographic manifestations after 14 days. The secondary outcome measures included remission rate or progression rate of critical illness， clinical remission rate of respiratory symptoms， routine blood examination， C-reactive protein （CPR） level， procalcitonin （PCT） level， and blood oxygen saturation （SPO2）. The safety was assessed based on liver and kidney functions and adverse events. ResultAfter the 14-day treatment， the ratio of patients showing improvement in the FHT group （100%） was higher than that in the control group （77.1%） （χ2=8.063，P<0.01）. The ratio of disease stages after treatment showed no significant difference between two groups. In the FHT group， the symptoms including cough， dyspnea， and fatigue were alleviated after treatment （P<0.01）. In the control group， the symptoms including fever， cough， and dyspnea were alleviated （P<0.01）， while the fatigue was not relieved after treatment. No significant difference was observed in the clinical symptoms between the two groups after treatment. After treatment， the FHT group showed decreased white blood cell （WBC） count and neutrophil-to-lymphocyte ratio （NLR） （P<0.01）， elevated platelet （PLT） level （P<0.05）， lowered CRP level （P<0.05）， and no significant difference in lymphocyte （LYM）， hemoglobin （Hb）， SPO2 or PCT level. The control group showed decreased NLR （P<0.05） and WBC count （P<0.01）， elevated PCT level （P<0.05）， and no significant change in LYM， Hb， PLT， SPO2 or CRP level after treatment. Furthermore， the FHT group had higher PLT level than the control group （P<0.05） after treatment， and other indicators had no significant differences between the two groups. The liver and kidney functions had no significant difference between the two groups after treatment. ConclusionFHT can safely promote the absorption of acute pulmonary inflammation in COVID-19 patients.

Clinical metabolomics research on Xiaozhang Tie with the umbilical compress therapy for cirrhotic ascites / 药学学报

We used metabolomics technology to identify and understand the biomarkers and therapeutic mechanisms of umbilical compress therapy based on Xiaozhang Tie (XT) to provide scientific evidence for its clinical application. A total of 10 patients with cirrhotic ascites and gastrointestinal motility disorders who were hospitalized in the Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine from November 2017 to June 2018 were divided into a placebo group (4 cases) or an XT group (5 cases), and 10 healthy volunteers were included as controls. This clinical trial was approved according to the Ethics Committee of Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine (2017-528-11-01). The patients in the XT group were given umbilical compress therapy with Xiaozhang Tie, and patients in the placebo group were administered a plaster patch in which the drug content was less than 5%, receiving one patch per day for three consecutive days. Non-targeted metabolomics technology and UPLC-Q/Orbitrap-MS/MS analysis technology were utilized to investigate the fluctuations in endogenous metabolic profiles in the patient's urine prior to and after administration of XT. By analyzing and comparing the urine metabolic profiles of patients with cirrhotic ascites to those of healthy volunteers, a total of 31 biomarkers were identified, 14 of which were significantly decreased by the intervention with Xiaozhang Tie (P <0.05). Pathway enrichment analysis revealed that phenylalanine metabolism and tryptophan metabolism are key pathways affected by XT treatment. The results suggest that XT can alleviate cirrhotic ascites by modulating abnormalities in amino acid metabolism.

Transcriptomics study of coronary slow flow disease and verification of differentially expressed genes / 中华心血管病杂志

Objective: To determine the differentially expressed genes between patients with coronary slow flow (SCF) and healthy controls, as well as to define the signal pathways and protein interactions related to the pathogenesis of the disease. Methods: A total of 43 hospitalized SCF patients in the year of 2018 and 43 healthy subjects, who underwent health checkup in the same year, were enrolled in this study. General data were obtained, blood samples were collected to determine the related indicators of glucose metabolism, lipid metabolism and kidney metabolism. RNA was extracted from blood monocytes, and the differential gene expression profiles were investigated by RNA-Seq. GO function annotation, KEGG pathway enrichment, protein interaction network analysis (PPI) and phenotype analysis were performed. The levels of related cytokines were detected by ELISA, and qPCR was used to verify differentially expressed genes of the two groups. Results: In the SCF group, there were 27 (62.79%) males and 16 females (37.21%), the average age was (54.3±8.8) years. In the control group, there were 29 males (67.44%) and 14 females (31.56%), the average age was (57.2±8.3) years. The percent of smoking history, abnormal fasting blood glucose, abnormal blood lipid levels and body mass index were significantly higher in the SCF group than in the control group (all P<0.01). There were 117 differentially expressed genes between SCF patients and healthy controls, of which 73 were up-regulated and 44 were down-regulated. Biological function analysis of the differentially expressed genes showed that these genes were mainly related to antigen processing and presentation, cell phagocytosis, immunoglobulin, intestinal immune network, Th1 and Th2 cell differentiation and Th17 cell differentiation pathways. The expression levels of inflammatory cytokines IL-6, IL-10, tumor necrosis factor-α and interferon-γ were significantly higher in SCF patients than in healthy controls (all P<0.05). Among the top 12 genes with the most significant differences between the two groups, qPCR analysis indicated consistent results with the transcriptome results in 11 out of 12 genes. PPI analysis showed that FPR2 and THBS3 proteins were at the core of the entire protein interaction network. Conclusion: Genes such as FPR2 and THBS3 may play important roles in the pathogenesis of SCF through immune-related pathways such as antigen processing and presentation and Th17 cell differentiation.

Advances in drug therapy for primary biliary cholangitis / 临床肝胆病杂志

Primary biliary cholangitis (PBC) is a cholestatic liver disease with unclear etiology and pathogenesis. Ursodeoxycholic acid and obeticholic acid are the therapeutic drugs for PBC approved by the US Food and Drug Administration, both of which have the effect of alleviating intrahepatic cholestasis. However, some patients still have poor biochemical response to them, and it is urgent to explore new therapeutic drugs or methods. Fibrates, immunosuppressants, and traditional Chinese medicine have a certain effect in the treatment of patients with PBC, but further studies are needed to analyze their clinical safety and mechanism of action.

Investigating Relationship Between Renal Damage and Endoplasmic Reticulum Stress in Diabetic Kidney Disease Rats with Blood Stasis Syndrome Based on Combination of Disease and Syndrome / 中国实验方剂学杂志

Objective:To observe the effect of blood stasis syndrome on renal damage and endoplasmic reticulum stress (ERS) in diabetic kidney disease (DKD) rats, and to explore the relationship between renal syndrome of blood stasis damage and ERS in DKD rats. Method:The 50 Male SD rats of SPF level were selected to establish DKD rat model by high fat and high sugar diet combined with intra-abdominal injection of streptozotocin(STZ). They were randomly divided into normal group, diabetes mellitus group and diabetes mellitus and blood stasis syndrome group（0.05 mg·kg-1）, among which diabetes mellitus and blood stasis syndrome group was prepared by dextran method. The general conditions, hemorheology indexes, 24 h urine protein, serum creatinine and renal pathology of the rats in each group were observed. Immunohistochemical analysis, Western blot and Real-time fluorescent quantitative polymerase chain reaction(Real-time PCR)were used to detect mRNA and protein expressions of endoplasmic reticulum stress-related proteins glucose regulated protein 78(GRP78), activating transcription factor-6(ATF6) and renal fibrosis index fibronectin(FN), and alpha smooth muscle actin(α-SMA). Result:Compared with normal group, the rats in diabetes mellitus group showed polyphagia, polyuria and weight loss, increased hemorheology index of whole blood viscosity and plasma viscosity(P<0.05，P<0.01), increased 24 h urine protein, serum creatinine and urea nitrogen(P<0.01), and increased renal pathology, and increased mRNA and protein expression of GRP78, ATF6, FN and α-SMA(P<0.05，P<0.01). After dextran preparation of blood stasis model. Diabetes mellitus and blood stasis syndrome group increased mortality, signs of change is more obvious in the diabetic group, whole blood viscosity, plasma viscosity, 24 h urine protein ration, serum creatinine and urea nitrogen were significantly higher than those in diabetic rats(P<0.01), pathological changes aggravated in the diabetes group. At the same time， mRNA and protein expressions of GRP78, ATF6, FN, and α-SMA in renal tissue were significantly higher than those in diabetic mellitus group(P<0.05，P<0.01). Conclusion:Under the combined disease and syndrome model, the blood stasis syndrome may further aggravate the pathological damage of the kidney of DKD rats, and is related to the enhancement of ERS in the kidney of DKD rats.

Effect of "Tongdu Tiaoshen" needling combined with swallowing training on dysphagia, cerebral blood flow and serum BDNF and NGF levels in ischemic stroke patients / 针刺研究

OBJECTIVE: To observe the efficacy of Tongdu Tiaoshen (dredging Governor Vessel and regulating mind) needling combined with swallowing training in the treatment of ischemic stroke (IS) with dysphagia, and to investigate its effect on cerebral blood flow and serum levels of brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF). METHODS: A total of 100 IS inpatients with dysphagia were enrolled in the present study, and randomly and equally divided into control group and treatment group by using a random number table. The patients of the control group received routine swallowing training including tongue extending and retracting, cheek-muscle training, breathing exercise, and laryngopharyngeal exercise, beginning from the 2nd day after hospitalization. The patients of the treatment group received manual acupuncture stimulation of Dazhui (GV14), Fengfu (GV16), Shenting (GV24), Shendao (GV11), Baihui (GV20), Shuigou (GV26), etc., on the basic treatment as those mentioned in the control group. The treatment was conducted once daily, 5 times per week and for 4 successive weeks. The swallowing ability was tested by using video fluoroscopic swallowing study (VFSS), Kubota water swallowing test, and the standard swallowing function assessment (SSA) scale, separately, and patients' daily life quality was assessed by using swallowing related quality of life questionnaire (SWAL-QOL). The cerebral hemodynamics including mean blood flow velocity (Vm), maximum peak flow speed (Vs), and vascular resistance index (RI) of the bilateral cerebral arteries was detected by transcranial color Doppler ultrasound tests, and serum BDNF and NGF contents were assayed by enzyme linked immunosorbent assay. The therapeutic effect of swallowing ability was evaluated after the treatment. RESULTS: After 4 weeks' treatment, the scores of Kubata water swallowing test, SSA, and SWAL-QOL and RI were considerably decreased (P<0.01), and the VFSS scores, Vs and Vm levels as well as serum BDNF and NGF contents were significantly increased (P<0.01) in both groups compared with their own pre-treatment. Of the 48 and 49 cases in the control and treatment groups, 4 (8.33%) and 8(16.33%) were cured, 26 (54.17%) and 33 (67.35%) effective, 18 (37.50%) and 8 (16.33%) failed, with the effective rate being 62.50% and 83.67%, respectively. The therapeutic effect of the treatment group was significantly superior to that of the control group in the effective rate (P<0.05), and in lowering the scores of Kubota water swallowing test, SWAL-QOL, SSA, RI, and in up-regulating the scores of VFSS, Vs and Vm, and serum BDNF and NGF levels (P<0.01). CONCLUSION: Tongdu Tiaoshen needling combined with swallowing training is effective in improving swallowing ability, promoting cerebral blood flow and in up-regulating serum neurotrophic factor levels in patients with dysphagia after ischemic stroke.

Short-term and long-term efficacy of PGLA thread-embedding therapy in treatment of stage Ⅰ post-stroke shoulder-hand syndrome / 针刺研究

OBJECTIVE: To investigate short-term and long-term efficacy of PGLA thread-embedding therapy in the treatment of stage Ⅰ post-stroke shoulder-hand syndrome. METHODS: A total of 60 patients with stage Ⅰ post-stroke shoulder-hand syndrome were randomly divided into control group and treatment group, with 30 patients in each group. The patients in the control group were given acupuncture combined with rehabilitation training, and those in the treatment group were given PGLA thread-embedding therapy combined with rehabilitation training. Conventional acupuncture was performed once a day, and thread-embedding therapy was performed once every 5 days. Both groups were treated for 30 days. Clinical outcome, Visual Analogue Scale (VAS) score, hand edema score, and Fugl-Meyer Assessment (FMA) score were compared within and between groups immediately and at 1 month after treatment. RESULTS: The treatment group had a significantly higher overall response rate than the control group immediately and at 1 month after treatment (93.33%/86.67% vs 73.33%/56.67%, P<0.05). Both groups had significant reductions in VAS score and hand edema score and a significant increase in FMA score immediately and at 1 month after treatment (P<0.05), and the treatment group had significantly greater improvements than the control group (P<0.05). CONCLUSION: PGLA thread-embedding therapy has a better clinical effect than conventional acupuncture in the treatment of stage Ⅰ post-stroke shoulder-hand syndrome.

Analysis of Mechanism of Artesunate in Treatment of Liver Cancer Based on Metabolomics / 中国实验方剂学杂志

Objective: To study on the antitumor mechanism of artesunate in the treatment of liver cancer based on gas chromatography-mass spectrometry (GC-MS). Method: CellTiter-Glo® Luminescent Cell Viability Assay was used to detect activity of artesunate with different concentrations (0, 12.5, 25, 50, 100 μmol·L-1) on human liver cancer Huh7, SMMC-7721 cells for 24, 48, 72 h. GC-MS was employed to analyze the changes of metabolites of artesunate in two kinds of hepatoma cells (Huh7, SMMC-7721) for 24 h. The data was preprocessed by Postrun Analysis 4.41 workstation. Partial least squares-discriminant analysis (PLS-DA) was used to analyze two sets of differential metabolites and to analyze metabolic pathways of differential metabolites based on MetaboAnalyst 3.0 software. Result: Compared with the normal group, after two kinds of liver cancer cells was treated by artesunate, a total of 39 identical metabolites in the cells have undergone significant changes, which were mainly related to five metabolic pathways,including biosynthesis of aminoacyl-transfer RNA (tRNA), metabolism of alanine, aspartic acid and glutamic acid, metabolism of glycine, serine and threonine, metabolism of arginine and proline, metabolism of glutathione. Conclusion: Artesunate (12.5-100 μmol·L-1) can inhibit the growth of liver cancer cells (Huh7, SMMC-7721), it mainly involves five metabolic pathways, which may be the pathway of artesunate against liver cancer.

Observation of the therapeutic and characteristic effects of terlipressin on refractory cirrhotic ascites / 中华肝脏病杂志

Objective@#To observe the therapeutic effect of terlipressin on refractory ascites (RA) in cirrhosis, and its role and impact on acute kidney injury (AKI).@*Methods@#A non-randomized controlled clinical trial data of 111 hospitalized cases of liver cirrhosis accompanied with RA was collected from Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Zhongshan Hospital of Hubei Province, The First Affiliated Hospital of Zhengzhou University, The First Affiliated Hospital of Medical School of Zhejiang University, and People's Hospital of Pudong New Area of Shanghai between March 2015 and March 2017. 26 cases of conventional treatment group (control group) were divided into two subgroups: RA without AKI (RA-NAKI) and RA with AKI (RA-AKI), and each subgroup consisted 13 cases. Patients with bacterial infection were treated with diuretics, albumin supplementation and antibiotics. 85 cases were presented in terlipressin combined treatment group, of which 27 cases were of RA-NAKI and 58 cases were of RA-AKI. Control group was injected terlipressin 1mg of intravenous drip or static push (once q6 h ~ 12 h) for more than 5 days. The treatment duration lasted for 2 weeks with 4 weeks of follow-up. Body weight, 24-hour urine volume, abdominal circumference, mean arterial pressure (MAP), liver and kidney function, anterior hepatic ascites, deepest point of ascites, and ultrasonographic detection of ascites in supine position before treatment, one and two weeks after treatment and 4 weeks after follow-up were compared. Count data were tested by χ 2. Samples of four groups at baseline were compared. One-way analysis of variance was used for normal distribution data and Kruskal-Wallis H test for non-normal distribution data. Repeated measures analysis of variance was used to compare the difference in efficacy between different time points before and after treatment in the group. The LSD method of one-way ANOVA was used to compare the two groups. A t-test of independent samples was used to compare the efficacy of different time series between the two groups. Mann-Whitney rank- sum test was used to compare the data of non-normal distribution between the two groups.@*Results@#(1) Baseline data were compared among 4 subgroups of terlipressin RA-NAKI and control RA-AKI. Control group age was higher than that of terlipressin group, and the serum creatinine (SCr) of the RA-AKI group was higher than RA-NAKI group, and there was no significant difference in the rest of the baseline data and the combined medication (P > 0.05). (2) An intra-group comparison between control and trelipressin before and after treatment showed that all patients had lower body mass, abdominal circumference and deepest ascites, and higher serum albumin (P < 0.05). 24-hour urine volume and MAP was significantly increased in the terlipressin group, while the pre-ascites, SCr and child Turcotte Pugh (CTP) scores were decreased. Body weight, abdominal circumference, pre-ascites, and deepest ascites of the terlipressin group were decreased, while MAP was increased during the treatment and follow-up periods. The differences were statistically significant when compared with the control group at the same time (P < 0.05). There was a statistically significant difference in the increase of 24-h urine volume in the terlipressin group compared with the control group (P < 0.05). The decrease in SCr and CTP in the terlipressin group after 2 weeks of treatment and 4 weeks of follow-up was statistically significant compared with the control group (P < 0.05). (3) Among the two subgroups of RA-AKI and RA-NAKI in the terlipressin group, the baseline SCr value of the former was higher than that of the latter. After treatment, the body weight, abdominal circumference, pre-ascites, deepest ascites and CTP scores were decreased. In the RA-AKI group, 24-hour urine volume, MAP, SCr and serum albumin concentration were significantly increased. The difference between the two subgroups before and after treatment was compared, and the body weight of RA-AKI group at 1, 2 weeks of treatment and 4 weeks of follow-up was significantly lower than RA-NAKI group, which were (- 2.3 ± 0.2 vs. - 1.5 ± 0.2) kg, (- 4.1 ± 0.2 vs. - 2.6 ± 0.2) kg, (- 4.2 ± 0.3 vs. - 2.4 ± 0.3) kg, respectively. RA-NAKI group urine volume was significantly increased at 2 weeks of treatment and 4 weeks of follow-up, which was (468 ± 42 vs. 110 ± 131) ml, (272 ± 34 ml vs. 11 ± 112) ml, respectively. SCr reduction (18.3 ± 4.7 vs. 0.9 ± 2.4) µmol/l at 4 weeks of follow-up was apparent in RA-NAKI group, and the difference was statistically significant (P < 0.05).@*Conclusion@#Addition of terlipressin to conventional treatment may significantly increase MAP, 24-h urine volume, improve renal function and promote ascites resolution in patients with refractory cirrhotic ascites. Moreover, its combination effect is more obvious in AKI patients, and adverse reactions are mild.

Research progress of terlipressin in the treatment of cirrhotic ascites-related complications / 中华肝脏病杂志

Ascites is the most common clinical symptom in the decompensated stage of patients with liver cirrhosis, and its co-existence in complex conditions, such as refractory ascites, hyponatremia, hepatorenal syndrome and other complications may harm seriously. Splanchnic vasodilation is the key pathological link of cirrhotic ascites-related complications and the treatment of this link can help to eliminate the symptoms of ascites and prevent the further deterioration of decompensated cirrhosis. This paper summaries the pharmacological basis, clinical evidence and application characteristics of a vasocative drugs-terlipressin in the treatment of cirrhotic ascites-related complications, and further analyzes the problems existing in the current research, then proposes a prospect.

Strategies for enhancing nanoscale brain targeting drug delivery / 药学学报

The blood brain barrier can selectively block the uptake of xenobiotics from peripheral blood into the brain. Although this is important for maintaining the stability of the brain environment and normal function of the central nervous system, it presents a challenge for delivery of therapeutic drugs to the brain. Passive brain-targeting drug carrier is able to increase the drug concentration in the brain by enhancing the affinity to blood-brain barrier and/or inhibiting the efflux absorbed drug via P-glycoprotein. The active brain-targeting drug carrier can be obtained by linking specific ligands or antibodies onto passive target carriers to achieve precise delivery of drugs to the brain. Dual targeting drug carriers obtained by combining tumor cell targeting with brain targeting have shown their advantages for treatment of brain tumors. The targeted drug delivery to brain will provide a unique manner for the treatment of brain diseases such as Alzheimer's, Parkinson's, brain tumors, and stroke. Among the drug delivery systems of passive brain-targeting, active brain-targeting and dual brain-targeting, we evaluated the strategies to improve brain drug delivery efficiency, such as by reducing carrier size, opening tight junctions between cells at the blood-brain barrier, incorporating hydrophilic groups on the surface of the carrier, and alternative intranasal drug administration.

Mitochondrial damage in early stage of pressure ulcer in rats / 基础医学与临床

Objective To investigate the mitochondrial damage and its effect in early stage of pressure ulcer in rats.Methods Forty rats were randomly divided into 5 groups(n=8), control group(Con group) rats without stress, the experimental group was treated with of 170 mmHg for 2 h and relax 0.5 h as one cycle(1C), experi-mental group was divided into 3C, 6C, 9C and 12C group.The pathological changes of the compressed muscle tissue of the rats in each group were observed by HE staining , Western blot was used to detect the expression of Bcl-2 and Bax in the compressed tissue , and the ultrastructure of muscle fibers and mitochondria were observed by transmission electron microscope .Results There were pathological damage and gradually increased in the ex-perimental groups, with the increase of compression cycle; the expression of Bcl-2 in each experimental group was significantly increased as compared with the control group(P<0.05), in the 3C group reached the peak, and then decreased; the expression of Bax was increased gradually with the increase of compression cycle ( P<0.05) , and in the 12C group reached the peak;with the increase of the compression cycle the muscle fibers of each experimental group appeared gradually increased pathological damage:disorder, dissolution and fracture, the ridge of the mitochondria disappeared, vacuolar degeneration, et al.Conclusions In the early stage of pres-sure ulcer in a rat , it brings occurred mitochondrial damage and induces apoptosis .

Effects of inhibiting of BAG-1 gene expression by RNA interference on prolifer-ation and apoptosis of cutaneous squamous cell carcinoma / 中国免疫学杂志

Objective:To investigate the effect of inhibiting of BAG-1 gene expression on proliferation and apoptosis of cutaneous squamous cell carcinoma.Methods:siRNA of BAG-1 was transfected in cutaneous squamous cell carcinoma A431 by Lipo-fectamineTM2000,cells were transfected for 48 h,the BAG-1 mRNA expression was detected by RT-PCR,BAG-1 protein expression was detected by Western blot;CCK8 and flow cytometry were used to detect the cell proliferation and apoptosis;Western blot was used to detect the expression of Bax,β-catenin and Survivin protein;IL-6 and VEGF content were detected by ELISA kit.Results:Compared with control group and negative control group,transfection of BAG-1 with siRNA could significantly inhibit the expression of BAG-1 at the transcriptional and translational levels;compared with the control group,the cells proliferation in BAG-1-siRNA group significantly decreased,apoptosis rate was significantly increased,Bax protein expression was up-regulated,expression of β-catenin,Survivin protein were down regulated,the contents of IL-6 and VEGF were decreased significantly(P<0.05).Conclusion:Silencing of BAG-1 expression can reduce the proliferation of cutaneous squamous cell carcinoma,promote apoptosis,up regulate Bax expression and down regulate the expression of Wnt/β-catenin signaling pathway,and reduce the secretion of IL-6 and VEGF factors.

Analysis of the risk factors, pathogenic bacteria and drug resistance of postoperative infection in patients with colorectal cancer / 新乡医学院学报

Objective To analyze the risk factors for postoperative infection of patients with colorectal cancer,and to investigate the distribution of pathogenic bacteria and drug resistance.Methods A total of 300 patients with colorectal cancer who underwent operation were selected from June 2013 to June 2017 in the Central Hospital of Tongchuan Mining Bureau,and the patients were divided into the infection group and the non-infection group according to the postoperative infection.The clinical data were compared between the two groups,and the risk factors for postoperative infection in colorectal cancer patients were analyzed by logistic regression.The total automatic bacterial culture apparatus was used for bacterial culture,and the drug sensitivity test was carried out by Kirby-Bauer.Results Among the 300 colorectal cancer patients,there were 61 cases of incision infection (infection group) and 239 cases without incision infection (non-infection group) after operation,the postoperative infection rate was 20.33％ (61/300).There were significant differences in the operation method,red blood cell counts,the levels of albumin,prealbumin and hemoglobin between the two groups (P ＜ 0.05);but there was no significant difference in sex,age,body mass index,diabetes mellitus history,abdominal surgery history,smoking history,drinking history,operation time,intraoperative bleeding volume,hospitalization time,indwelling time of urethral catheter,white blood cells and C-reactive protein levels between the two groups(P ＞ 0.05).Multiple factor logistic regression analysis showed that the decrease of albumin and prealbumin level was the independent risk factor for postoperative infection in patients with colorectal cancer (P ＜0.05).A total of 52 strains of pathogenic bacteria were detected in 61 cases of colorectal cancer combined with postoperative infection,including 38 strains of gram negative bacteria (73.08％),10 strains of gram positive bacteria (19.23％) and 4 strains of fungi (7.69％).Escherichia coli,Klebsiella pneumoniae,Enterobacter cloacae,Pseudomonas aeruginosa had high drug resistance to ampicillin,cefaclor,ceftriaxone,ceftizoxime,cefepime,cefoperazone,ceftazidime and ofloxacin;but they had low drug resistance to piperacillin,imipenem and amikacin.Conclusion The incidence of postoperative infection in patients with colorectal cancer is higher,the decrease of albumin and prealbumin is the independent risk factor for postoperative infection in patients with colorectal cancer.Gram-negative bacteria are the main pathogens of postoperative infection in patients with colorectal cancer and have high resistance to common antibiotics.

Antipyretic Effect of Herba Ephedrae-Ramulus Cinnamomi Herb Pair on Yeast-Induced Pyrexia Rats: A Metabolomics Study / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the antipyretic mechanism of Herba Ephedrae (Eph)-Ramulus Cinnamomi (RC) herb pair on yeast-induced pyrexia in rats.</p><p><b>METHODS</b>Totally 30 qualified male SD rats were randomly assigned to the normal control (NC) group, the pyrexia model (model) group, the Eph, RC and Eph-RC treatment groups by a random digital table, 6 rats in each group. Each rat received a 20% aqueous suspension of yeast (10 mL/kg) except the NC group. The 3 treatment groups were administered 8.1, 5.4 and 13.5 g/kg Eph, RC and Eph-RC respectively at 5 and 12 h after yeast injection, the NC group and the model groups were administered equal volume of distilled water. Rectal temperatures were measured at 0, 6, 8, 10, 12, 15, 18, 24 and 30 h and urine was collected prior to yeast injection and at 6, 10, 18, 24, 30, and 36 h after yeast injection. Then urine metabolomic profiling by gas chromatography tandem mass spectrometry, coupled with multivariate statistical analysis and pattern recognition techniques were used to explore the antipyretic effects of Eph-RC. Partial least squares discriminate analysis was used to analyze the metabolomics dataset including classification and regression in metabolomics plot profiling.</p><p><b>RESULTS</b>Compared with the NC group, rectal temperatures were significantly higher in the model group (P<0.01), while 3 treatment groups decreased significantly compared with the model group (P<0.05 or P<0.01). Rectal temperatures of Eph-RC-treated rats started to go down at 6 h, and markedly decreased at 8, 12, 15, 18 and 24 h (P<0.05 or P<0.01), while those of the Eph and RC groups had decreased firstly at 8 h and were markedly lower at 12 h (P<0.05 or P<0.01). Seventeen potential biomarkers related to pyrexia were confirmed and identified, including pyruvic acid, L-phenylalanine, L-tyrosine, phenylacetic acid, hippuric acid, succinic acid, citrate and so on. Eight potential alterations of metabolic pathways including phenylalanine metabolism, citrate cycle, tryptophan metabolism, biosynthesis of valine, leucine and isoleucine, were identified in relation to the antipyretic effects of Eph-RC using MetPA software.</p><p><b>CONCLUSION</b>The antipyretic effect of Eph-RC herb pair on yeast-induced pyrexia in rats involved correction of perturbed amino acid, fatty acid, and carbohydrate metabolism according to the metabolic pathway analysis with MetPA.</p>

Correlation of liver stiffness measured by FibroTouch and FibroScan with Ishak fibrosis score in patients with chronic hepatitis B / 中华肝脏病杂志

Objective@#To investigate the correlation of liver stiffness measured by FibroTouch (FT) and FibroScan (FS) with Ishak fibrosis score in patients with chronic hepatitis B.@*Methods@#A total of 313 patients with chronic hepatitis B who visited Department of Liver Cirrhosis in Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine from November 2014 to May 2016 were enrolled. All the patients underwent liver biopsy, and FT and FS were used to determine liver stiffness measurement (LSM). Serum biochemical parameters were measured, and the aspartate aminotransferase-to-platelet ratio index (APRI) in a multi-parameter model of liver fibrosis and fibrosis-4 (FIB-4) index were calculated. The consistency between the results of four noninvasive examinations and Ishak fibrosis score was compared. The t-test was used for comparison of LSM determined by FT and FS. Pearson correlation analysis was used investigate the correlation between LSM determined by FT and FS; Spearman correlation analysis was used to investigate the correlation of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels and Knodell score with LSM determined by FT and FS; the correlation between LSM determined by FT and FS and fibrosis stage was analyzed by partial correlation analysis adjusted by Knodell score for liver inflammatory activity; Spearman correlation analysis was used for APRI, FIB-4, and fibrosis stage. Based on the Ishak fibrosis score, the receiver operating characteristic (ROC) curve was used to analyze the values of four noninvasive methods in the diagnosis of liver fibrosis.@*Results@#There was no significant difference in LSM measured by FT and FS in all patients (15.75±9.42 kPa vs 15.42±10.52 kPa, P > 0.05) and Pearson correlation analysis indicated a significant positive correlation between them (r = 0.858, P < 0.01); serum ALT and AST levels and liver inflammatory activity were correlated with LSM determined by FT and FS. There was a significant positive correlation between LSM determined by FT and FS and fibrosis stage (r = 0.501 and 0.526, both P < 0.001), and APRI and FIB-4 were also positively correlated with fibrosis stage (r = 0.236 and 0.218, both P < 0.001). Based on the Ishak fibrosis score, in the diagnosis of fibrosis stages F3, F4, F5, and F6, the areas under the ROC curve were 0.915/0.856/0.839/0.816 for FT, 0.933/0.883/0.849/0.856 for FS, 0.618/0.630/0.608/0.638 for APRI, and 0.614/0.624/0.595/0.649 for FIB-4, and FT and FS had a significantly larger areas under the ROC curve than APRI and FIB-4.@*Conclusion@#LSM determined by FT or FS has a good correlation with the Ishak fibrosis score, so FT and FS have a significantly better diagnostic performance for liver fibrosis than APRI and FIB-4.

Antipyretic and anti-asthmatic activities of traditional Chinese herb-pairs, Ephedra and Gypsum / 中国结合医学杂志

<p><b>OBJECTIVE</b>Mahuang-Shigao herb-pair is a famous formula composed of Ephedra and Gypsum. The herb-pair is frequently used for treating cold symptoms and bronchial asthma in the clinical practice of Chinese medicine (CM). In the present study, we evaluated evidence for the benefit of combined use of Ephedra and Gypsum by analyzing the antipyretic and anti-asthmatic activities of Ephedra-Gypsum.</p><p><b>METHODS</b>The antipyretic effects of Ephedra-Gypsum were evaluated in yeast-induced hyperthermia test. Thirty male Wistar rats were randomly divided into 5 groups, including control group, standard aspirin group, and 3 Ephedra- Gypsum groups of different doses (6, 12, 24 g/kg). Ephedra-Gypsum extract and asprin were administered orally 6 h after the injection of yeast solution and body temperature was measured every 1 h for 8 h. The antiasthmatic effects of Ephedra-Gypsum were evaluated using an ovalbumin (OVA)-induced asthmatic rat model. Thirty-six male SD rats were randomly divided into 6 groups. Rats were alternately sensitized and OVA+Al(OH) challenged by exposure to mists of ovalbumin. Ephedra-Gypsum extracts (6, 12, 24 g/kg) or dexamethasone were administered 45 min prior to the allergen challenge for 8 days. Latent period and the weight of wet to dry ratio of lung were determined. In addition, the eosinophils in blood and white blood cell (WBC) were counted by an YZ-Hemavet Analyzer.</p><p><b>RESULTS</b>The Ephedra-Gypsum extracts at test dose (6, 12, 24 g/kg) significantly and dose-dependently attenuated yeast-induced fever in rats. The Ephedra-Gypsum extracts also prolonged the latent period, reduced OVA-induced increases in eosinophils and WBC, and decreased the wet and dry weight ratio of the lungs in the anti-asthmatic test.</p><p><b>CONCLUSIONS</b>These findings indicate that the Ephedra-Gypsum extract has antipyretic and anti-asthmatic properties. Hence, the results support additional scientific evidence in prescriptions.</p>