Objective To investigate the role of long non-coding RNA(lnc RNA)ZFAS1 in glioma cells'sensitivity to cisplatin and its underlying mechanisms.Methods By analyzing the knockdown of ZFAS1 on the sensitivity of glioma cells to cisplatin using real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)experiments,and the cells were divided into sh-NC group(transfected with sh-NC lentiviral plasmid),sh#1 group(transfected with sh-ZFAS1-1 lentiviral plasmid)and sh#2 group(transfected with sh-ZFAS1-2 lentiviral plasmid).Dual luciferase experiments verified the interaction between ZFAS1 and miR-193b-3p,and the cells were divided into ZFAS1-WT+NC inhibitor group(transfected with ZFAS1 wild-type plasmid and NC inhibitor),ZFAS1-WT+miR-193b-3p inhibitor group(transfected with ZFAS1 wild-type plasmid and miR-193b-3p inhibitor),ZFAS1-Mut+NC inhibitor group(transfected with ZFAS1 mutant plasmid and NC inhibitor)and ZFAS1-Mut+miR-193b-3p inhibitor group(transfected with ZFAS1 mutant plasmid and miR-193b-3p inhibitor).Cell counting kit-8(CCK-8)and terminal deoxynucleotidly transferase mediated labeling(TUNEL)experiments were used to analyze the effect of ZFAS1/miR-193b-3p on the sensitivity of glioma cells to cisplatin,and the cells were divided into blank control group(0 μg·mL-1 cisplatin treatment of U251 cells),0.5 μg·mL-1 cisplatin+sh-NC+NC inhibitor group(0.5 μg·mL-1 cisplatin treatment of U251 cells co-transfected with sh-NC lentiviral plasmid and NC inhibitor),0.5 μg·mL-1 cisplatin+sh#1+NC inhibitor group(0.5 μg·mL-1cisplatin treatment of U251 cells co-transfected with sh-NC lentiviral plasmid and NC inhibitor),and 0.5 μg·mL-1 cisplatin+sh#1+miR-193b-3p inhibitor group(0.5 μg·mL-1 cisplatin treatment of U251 cells co-transfected with sh-ZFAS1-1 lentiviral plasmid and miR-193b-3p inhibitor).Results The results of the experiment showed that the expression levels of ZFAS1 in the sh-NC group,sh#1 group and sh#2 group were 1.00±0.17,0.48±0.06 and 0.68±0.08.The fluorescence activities of ZFAS 1-WT+NC inhibitor group,ZFAS1-WT+miR-193b-3p inhibitor group,ZFAS1-Mut+NC inhibitor group and ZFAS1-Mut+miR-193b-3p inhibitor group were 1.00±0.10,1.45±0.11,1.02±0.09 and 0.97±0.13.The proliferation rates at 72 h for the blank control group,0.5 μg·mL-1 cisplatin+sh-NC+NC inhibitor group,0.5 μg·mL-1 cisplatin+sh#1+NC inhibitor group and 0.5 μg·mL-1cisplatin+sh# 1+miR-193b-3p inhibitor group were(100.00±14.13)％,(96.62±9.82)％,(60.56±6.08)％and(78.64±7.22)％;while the apoptosis rates at 72 h were(9.52±1.11)％,(10.12±1.34)％,(16.08±1.52)％and(12.22±1.19)％.Comparied between blank control group and 0.5 μg·mL-1 cisplatin+sh-NC+NC inhibitor group,0.5 μg·mL-1 cisplatin+sh#1+NC inhibitor group and 0.5 μg·mL-1 cisplatin+sh # 1+miR-193b-3p inhibitor group,the differences were statistically significant(all P＜0.05).Conclusion This study reveals the important role of ZFAS1 in cisplatin sensitivity in glioma and elucidates its mechanism of influencing drug sensitivity through the regulation of miR-193b-3p.

ObjectiveTo investigate the therapeutic effect and the possible mechanism of Mankuining decoction on dextran sulfate sodium （DSS）-induced ulcerative colitis （UC） in mice. MethodA total of 90 male SPF C57BL/6 mice were randomly classified into normal group， model group， mesalazine group （0.266 g·kg^-1）， and high-， and medium-， low-dose （20， 10， 5 g·kg^-1） Mankuining decoction groups， with 15 rats in each group. Mice， except the normal group， drank 3% DSS solution for 7 days to induce UC， and administration （ig） started on the day of modeling. The model group and the normal group were given equivalent amount of 0.9% normal saline once a day for 7 days. The general conditions of mice were recorded every day and the disease activity index （DAI） was calculated. On the 8^th day， mice were killed by cervical dislocation. All the colons and feces were collected. The length of colon was recorded， and the histopathological changes of colon were observed based on hematoxylin-eosin （HE） staining. The content of inflammatory factors in colon was detected by enzyme-linked immunosorbent assay （ELISA）， and the changes of intestinal flora in mouse feces were determined based on 16SrRNA sequencing. ResultCompared with the normal group， the model group had severe colon damage， reduction in colon length （P<0.01）， increase in DAI （P<0.01）， decrease in interleukin-10 （IL-10） and transforming growth factor-β₁ （TGF-β₁） in colon（P<0.01）， rise of interleukin-1β （IL-1β）， interleukin-6 （IL-6）， interleukin-17α （IL-17α）， and tumor necrosis factor-α （TNF-α） in colon （P<0.05， P<0.01）， and decrease in abundance and diversity of intestinal flora. Compared with the model group， mesalazine and high-， medium-， low-dose Mankuining decoction alleviated the colon injury， recovered the length of colon （P<0.01）， decreased DAI （P<0.01）， increased IL-10 and TGF-β₁ in colon （P<0.01）， and decreased IL-1β， IL-6， IL-17α， and TNF-α in colon （P<0.01）. Moreover， they raised the abundance and diversity of intestinal flora compared with the model group， as manifested by the increase in the abundance of Firmicutes， Akkermansia， Dubosiella， and Blautia and the decrease in the abundance of Bacteroidetes， Muribaculaceae， Clostridia_UCG-014， and Alistipes. ConclusionMankuining decoction has definite effect in treating UC mice， and the effect is positively correlated with the concentration. In addition， different concentration has different influence on the structure of flora. The mechanism is the likelihood that it alleviates the disorder of intestinal flora to restore intestinal immune balance and further promote the recovery of colonic mucosa.

Objective: Jiaotaiwan is a classic prescription in traditional Chinese medicine for insomnia. Modern clinical research has proved its anti-diabetes effect by "the same treatment for different diseases" theory, so it is necessary to study its pharmacological mechanism for anti-diabetes effect. Method: In this study, the integrative pharmacology platform of traditional Chinese medicine (TCMIP) was used to explore the potential target and mechanism of Jiaotaiwan, and construct its core target network for diabetes. Then the enrich analysis of GO and KEGG on key targets was conducted to build the visual multilayer association network of "Jiaotaiwan-active composition-core target-key pathway". Result:28 active ingredients were obtained from Jiaotaiwan in this study. Its anti-diabetes effect was relevant to 187 core targets,including 15 known disease targets such as vasopressin V2 receptor (AVPR2), receptor activity-modifying protein 1 (RAMP1), receptor activity-modifying protein 3 (RAMP3), insulin receptor (INSR), and insulin-like growth factor 1 receptor (IGF1R); as well as 71 predictive drug targets such as cyclin-dependent kinase 9 (CDK9), glucokinase (GCK), NF-kappa-B inhibitor alpha (NFKBIA), NF-kappa-B p100 subunit (NFKB2), and hypoxia inducible factor-1 alpha (HIF1A). Conclusion:The anti-diabetes mechanism of Jiaotaiwan may be associated with activation of adenylate cyclase activity, cellular response to glucagon stimulus, activation of mitogen-activated protein kinase (MAPK) activity, endocrine system, gonadotropin-releasing hormone (GnRH) signaling pathway, Chemokine signaling pathway, phosphatidylinositol 3-kinase-serine/threonine kinases (PI3K-Akt) signaling pathway and other related biological processes and pathways. This study provides a scientific evidence for further study of the anti-diabetes mechanism of Jiaotaiwan.

The anhydrite and gypsum are the main sulfate minerals during evaporation of seawater or lake.They record the information about relative hydrogeology and the composition of mother liquor.Boron is diffluent element, and often occurs in all kinds of evaporites.Presently, the boron isotope has been applied widely in mineral deposits forming, geochemistry and palaeoenvironment.However, there is little research about characteristic of boron isotope in anhydrite and gypsum minerals, because of the low content of boron and micro-solubility in water and hydrochloric acid.This study developed a method of extracting and purifying boron in anhydrite and gypsum by phase transformation and ion-exchange.Firstly, the samples were mixed with ammonium hydrogen carbonate to transform the calcium sulfate to calcium carbonate.And diluted hydrochloric acid (1 mol/L) was added to resolve calcium carbonate.The percent conversion was about 85%in the first stage, and up to complete resolution by repeating this process.Secondly, boron specific ion-exchange resin ( Amberlite IRA 743 ) was used to gather the boron ions fully and further refined the samples with more than 1 μg of boron by anionic and cationic resin mixed by Ion Exchange Ⅱ and Dowex 50 W × 8.Finally, according to the modified method by He, the values of boron isotope were determined by TIMS.The boron content is analytically pure gypsum was 3.501 ± 0.128 μg/g ( n=12 , RSD=3.6%) and the average recovery was 100.47%.Besides, the δ11B value of analytically pure gypsum added with NIST SRM 951 was 17.98‰±0.21‰ (n=3, RSD=1.2%).This method has good repeatability and can meet the requirements of boron isotopic measurement of anhydrite and gypsum.

BACKGROUND:During the percutaneous vertebroplasty, the optimal dose of bone cement that can bring favorable cement dispersion and remodel the biomechanical balance of the fractured vertebrae remains controversial. OBJECTIVE:To investigate the dispersion degree of small dose of bone cement in vertebroplasty. METHODS: In this experiment, 18 sheep selected with the same condition were randomly divided into three groups (group A, group B, group C), 6 in each group. A model of thoracolumbar vertebral compression fracture (T12, L1, L2) was made in each sheep. The injected volume of bone cement in groups A, B, C was 15％, 20％, 25％ of the average volume of adjacent vertebral bodies, respectively. Postoperative CT images were used to evaluate the bone cement dispersion. Dispersion degree of bone cement among the three groups was compared by the Kruskal-Wallis test. RESULTS AND CONCLUSION:There was no statistical difference in the dispersion degree of bone cement among the three groups, and the excellent and good rate of dispersion was over 80％. To conclude, the optimal dose of bone cement injected into the fractured vertebra is 15％ of the average volume of adjacent vertebral bodies, which can achieve good dispersion degree and restore the biomechanical stability of the vertebral body.

BACKGROUND: Currently, there are few studies about prostaglandin E1 in the paracrine and migration of bone marrow mesenchymal stem cells. OBJECTIVE: To explore the effects of prostaglandin E1 in the paracrine and migration of bone marrow mesenchymal stem cells. METHODS: Bone marrow mesenchymal stem cells isolated from Sprague Dawley rats were cultured in vitro. Passage 3 cells were co-cultured with prostaglandin E1 at concentrations of 10 μg/L, and then culture supernatant was collected at 3, 6, 9, 12, 24, 48, and 72 hours after co-culture. The level of vascular endothelial growth factor was detected by enzyme-linked immunosorbent assay. Effects of prostaglandin E1 on the migration of bone marrow mesenchymal stem cells were detected by Transwell assay and cell scratch assay. RESULTS AND CONCLUSION: After treatment with prostaglandin E1 for 3 hours, bone marrow mesenchymal stem cells began to secrete vascular endothelial growth factors, and the secretion level was peaked at 24 hours and then gradually decreased. Results from the Transwell assay and cell scratch assay showed that the migration ability of bone marrow mesenchymal stem cells was significantly promoted by prostaglandin E1 (P < 0.05). Overall findings reveal that prostaglandin E1 promotes the secretion of vascular endothelial growth factor from bone marrow mesenchymal stem cells and enhances cell migration.

BACKGROUNDIntermittent hypoxia (IH) is a key element of obstructive sleep apnea (OSA) that can lead to disorders in the liver. In this study, IH was established in a rat model to examine its effects on the expression of hepatic cytochrome P450 (CYP) and CYP regulators, including nuclear receptors.

METHODSHematoxylin and eosin staining was conducted to analyze the general pathology of the liver of rats exposed to IH. The messenger RNA (mRNA) expression levels of inflammatory cytokines, CYPs, nuclear factor-κB (NF-κB), and nuclear factors in the liver were measured by quantitative reverse transcription polymerase chain reaction.

RESULTSWe found inflammatory infiltrates in the liver of rats exposed to IH. The mRNA expression level of interleukin-1beta was increased in the liver of the IH-exposed rats (0.005 ± 0.001 vs. 0.038 ± 0.008, P = 0.042), whereas the mRNA expression level of Cyp1a2 was downregulated (0.022 ± 0.002 vs. 0.0050 ± 0.0002, P = 0.029). The hepatic level of transcription factor NF-κB was also reduced in the IH group relative to that in the control group, but the difference was not statistically significant and was parallel to the expression of the pregnane X receptor and constitutive androstane receptor. However, the decreased expression of the glucocorticoid receptor upon IH treatment was statistically significant (0.056 ± 0.012 vs. 0.032 ± 0.005, P = 0.035).

CONCLUSIONSThese results indicate a decrease in expression of hepatic CYPs and their regulator GR in rats exposed to IH. Therefore, this should be noted for patients on medication, especially those on drugs metabolized via the hepatic system, and close attention should be paid to the liver function of patients with OSA-associated IH.

Objective To explore the therapeutic effects of endovenous laser ablation (EVLA) combined with percutaneous continuous circumsuture (PCCS) and EVLA in treating severe great saphena varicose. Methods A total of 60 patients with unilateral great saphenous varicose level C5-C6 were randomly divided into control group and experimental group according to the CEAP system. Control group was given EVLA surgery while experimental group was given EVLA+PCCS surgery. Data of operation time, hospital stay, intraoperative blood loss, the rate of ulcer healing, variceal recurrence rate and postoperative complication rate within 6 months after operation were compared between two groups. Results The mean operative time and intraoperative blood loss were lower in the experimental group than those in the control group ( P<0.05). There were no significant differences in hospital stay, ulcer healing rate and recurrence rate between two groups ( P>0.05). No deep venous thrombosis was found after treatment in two groups. The occurrence rates of skin burns and subcutaneous ecchymosis were significantly lower in the experimental group than those in the control group ( P<0.05). There were no significant differences in the incidence rates of other complications between two groups (P>0.05). Conclusion EVLA combined with PCCS in the treatment of severe saphenous varicose veins can significantly shorten the operation time, reduce the amount of bleeding, reduce the incidence rates of skin burns and subcutaneous ecchymosis on the premise of promising cure rate and recurrence rate. Overall, the combination therapy is superior than monotherapy.

Background: Intermittent hypoxia (IH) is a key element of obstructive sleep apnea (OSA) that can lead to disorders in the liver. In this study, IH was established in a rat model to examine its effects on the expression of hepatic cytochrome P450 (CYP) and CYP regulators, including nuclear receptors. Methods: Hematoxylin and eosin staining was conducted to analyze the general pathology of the liver of rats exposed to IH. The messenger RNA (mRNA) expression levels of inflammatory cytokines, CYPs, nuclear factor?κB (NF?κB), and nuclear factors in the liver were measured by quantitative reverse transcription polymerase chain reaction. Results: We found inflammatory infiltrates in the liver of rats exposed to IH. The mRNA expression level of interleukin?1beta was increased in the liver of the IH?exposed rats (0.005 ± 0.001 vs. 0.038 ± 0.008, P = 0.042), whereas the mRNA expression level of Cyp1a2 was downregulated (0.022 ± 0.002 vs. 0.0050 ± 0.0002, P = 0.029). The hepatic level of transcription factor NF?κB was also reduced in the IH group relative to that in the control group, but the difference was not statistically significant and was parallel to the expression of the pregnane X receptor and constitutive androstane receptor. However, the decreased expression of the glucocorticoid receptor upon IH treatment was statistically significant (0.056 ± 0.012 vs. 0.032 ± 0.005, P = 0.035). Conclusions: These results indicate a decrease in expression of hepatic CYPs and their regulator GR in rats exposed to IH. Therefore, this should be noted for patients on medication, especially those on drugs metabolized via the hepatic system, and close attention should be paid to the liver function of patients with OSA?associated IH.

Objective To obtain monoclonal antibodies ( mAbs ) against neutrophil gelatinase-associated lipocalin ( NGAL ) and a chemiluminescense immune quantification assay based one paired mAbs.Methods Six-to-eight weeks old female BALB/c mice were immunized with the purified recombinant human NGAL antigen( rhNGAL) that was produced by the Escherichia coil expression system.The spleen was fused with hybridoma for screening anti-NGAL monoclonal antibodies by indirect ELISA.Western blot was implemented to identify the reactivity with native NGAL. Results The rhNGAL antigen was found to form disulfide cross-linked dimers and present excellent immunogenicity.The reaction titer of the immune serum of NGAL immunized mice was about 106.Thirty mAbs were screened by indirect ELISA, hereinto;the EC50 values of mAb23C12 and 38D10 were 0.034 g/mL, 0.022 g/mL respectively.The antibodies pair, 38D10/23C12-SAE labeled with AcridiniumEster(AE), were shown to work well in chemiluminescense immune response quantitative detection which was screened by NGAL standardand clinical urine samples.This detection can resolve positive and negative samples with a statistically significant difference (P<0.0001).And the correlation coefficient R2between NGAL quantitative results and that of the Abbott's NGAL chemiluminescence immune assay kit was greater than 0.97.The detection linear range was 10-1500 ng/mL, analytical sensitivity of the method was 0.63 ng/mL.Conclusion Highly purified rhNGAL antigen and specific anti-NGAL monoclonal antibodies are generated in this study.The detection capability of method is comparable with that of the international commercial kit.