Objective:To discuss whether safranal affects the proliferation,migration,and phenotypic transformation of the vascular smooth muscle cells(VSMCs)in a high-glucose environment and to clarify the function of safranal in the prevention and treatment of diabetic(DM)vascular complications.Methods:The SD rats were selected as experimental subjects;primary VSMCs were cultured from rat thoracic aortas and divided into control group,25 mmol·L-1 high glucose(HG)group,HG+20 μmol·L-1 safranal group,HG+40 μmol·L-1 safranal group,and HG+80 μmol·L-1 safranal group.The cells in control group received no treatment;the cells in 25 mmol·L-1 HG group were pretreated with 25 mmol·L-1 HG;the cells in HG+20,40,and 80 μmol·L-1 safranal groups were further treated with 20,40,and 80 μmol·L-1 safranal respectively for 48 h on the basis of 25 mmol·L-1 HG group.Cell counting kit-8(CCK-8)method was used to determine the appropriate concentration of safranal and detect the viabilities of the VSMCs in various groups;cell scratch healing assay was used to detect the scratch healing rates of the VSMCs in various groups;Transwell chamber assay was used to detect the numbers of the migration VSMCs in various groups;immunofluorescence method was used to detect the fluorescence intensities of alpha-smooth muscle actin(α-SMA)and rabbit anti-osteopontin(OPN)in the VSMCs in various groups;Western blotting method was used to detect the expression levels of OPN,α-SMA,and proliferating cell nuclear antigen(PCNA)in the VSMCs in various groups.Results:Under microscope,on the 4th day of in vitro culture,the spindle-shaped or triangular cells crawled out from the edge of the thoracic aorta tissue blocks,with long spindle being the most common morphology.On the 14th,the cells gradually covered the bottom of the dish;when cell density reached 80%-90%,the characteristic"hills and valleys"growth pattern appeared.Third-generation cells were taken for immunofluorescence identification;immunofluorescence staining with VSMC-specific marker α-SMA showed positive expression of α-SMA protein in the primarily cultured VSMCs.The CCK-8 assay results showed that compared with control group,the cell viability of the cells in 160 μmol·L-1 safranal group was significantly decreased(P<0.01),indicating toxic damage to the cells.Under the conditions of safranal concentrations at 20,40,and 80 μmol·L-1 respectively,after 48 h intervention on VSMCs,no significant adverse effect on cell viability was observed;considering both the effect and toxicity of safranal,these three concentrations were used in subsequent cell experiments.After 48 h intervention,compared with control group,the activity of the VSMCs in 25 mmol·L-1 HG group was increased(P<0.001);compared with 25 mmol·L-1 HG group,the activities of the VSMCs in HG+20,40,and 80 μmol·L-1 safranal groups were gradually decreased(P<0.05).The cell scratch healing assay and Transwell assay results showed that after 48 h intervention,the scratch healing rate of the VSMCs in 25 mmol·L-1 HG group was significantly higher than that in control group(P<0.01),and the number of transmembrane cells through the Transwell chamber was significantly increased(P<0.05);compared with 25 mmol·L-1 HG group,the scratch healing rates of the VSMCs in HG+20,40,and 80 μmol·L-1 safranal groups were gradually decreased(P<0.05),and the number of transmembrane cells was decreased(P<0.05).The immunofluorescence staining results showed that compared with control group,the fluorescence intensity of α-SMA protein in the VSMCs in 25 mmol·L-1 HG group was significantly weakened(P<0.001),while the fluorescence intensity of OPN protein was significantly enhanced(P<0.001);compared with 25 mmol·L-1 HG group,the fluorescence intensities of α-SMA protein in the VSMCs in HG+20,40,and 80 μmol·L-1 safranal groups were gradually increased(P<0.05),and the fluorescence intensities of OPN were gradually weakened(P<0.05).The Western blotting method results showed that compared with control group,the expression level of α-SMA protein in the VSMCs in 25 mmol·L-1 HG group was decreased(P<0.05),and the expression levels of PCNA and OPN proteins were increased(P<0.01);compared with 25 mmol·L-1 HG group,the expression level of α-SMA protein in the VSMCs in HG+20,40,and 80 μmol·L-1 safranal groups were increased(P<0.05),and the expression levels of PCNA and OPN proteins were decreased(P<0.05).Conclusion:Safranal can inhibit the proliferation,migration,and phenotypic transformation of the VSMCs induced by high glucose.

TetR family transcriptional regulators (TFRs) are widely distributed in bacteria and archaea, and the first discovered TFR was confirmed to control the expression of tetracycline efflux pump in Escherichia coli. TFRs can bind DNAs and ligands. Small molecule ligands can induce conformational changes of TFRs, inhibiting or promoting TFRs to control target gene expression. Currently, TFRs have a wide variety of ligands, including carbohydrates, proteins, fatty acids and their derivatives, metal ions, and so on. Due to the diversity of ligands, TFRs regulate a wide range of physiological processes, from basic carbon metabolism and nitrogen metabolism to quorum sensing and antibiotic biosynthesis. On the basis of the recent studies in our laboratory and the literature, we review here the regulatory mechanism mediated by ligands of TFRs in primary and secondary metabolism, as well as the application of ligands for TFRs in the development of gene route and the activation of antibiotic biosynthesis.
Anti-Bacterial Agents ; Bacteria/metabolism* ; Bacterial Proteins/metabolism* ; Gene Expression Regulation, Bacterial ; Ligands ; Quorum Sensing

Objective To investigate the effect of G protein?coupled estrogen receptor (GPER) on the diastolic function of renal interlobular artery and reduce renal ischemia?reperfusion injury in rats. Methods Female ovariectomized rats were divided into control group; ischemia?reperfusion injury (IRI) group;GPER?specific agonist (G1) intervention group;GPER?specific blocker+GPER?specific agonist (G15+G1) intervention group. Histopathological examination (HE staining), renal function test and Paller score were used to identify the success of the model and the degree of kidney damage. In vitro microvascular pressure diameter measuring instrument was used to detect the relaxation and contraction activity of renal interlobular artery in each group. Immunofluorescence technique was used to observe the expression of GPER on the renal interlobular artery. Westernblotting was used to detect the expression of GPER protein in renal interlobular artery of rats in each group. The NO content was determined by a nitrate reductase method. Results Compared with IRI group, serum BUN, Scr level and Paller score in G1 intervention group were significantly decreased (all P<0.05). The systolic rate of renal interlobar artery was significantly increased [(40.76 ± 1.57)％ vs (29.78 ± 1.87)％, P<0.05]. The results of immunofluorescence showed that GPER was expressed in renal interlobular artery smooth muscle cells and endothelial cells, and the expression of IRI group was higher than that of the control group. The expression of G15+G1 intervention group was lower than that of G1 intervention group (all P<0.05). Compared with the IRI group, the NO content in the G1 intervention group increased significantly (all P<0.05). Conclusions During renal ischemia ?reperfusion injury, GPER may regulate the systolic and diastolic activity of the renal interlobar artery by increasing the content of NO, so as to alleviate the renal ischemia?reperfusion injury.

Objective To explore the significance of abnormal lipid metabolism induced by hypoxia in mice with pulmonary hypertension.Methods Thirty male C57BL/6 mice were chosen and were randomly divided into the model group and the control group, each consisting of 15 animals.The mice in the model group were exposed to chronic hypoxia for the development of hypoxic pul -monary hypertension model , and the mice in the control group were housed in the chamber at the normal ambient air .Right heart cathe-terization was used to measure right ventricular systolic pressure in the 2 groups, Masson method was used to observe the small pulmona-ry vascular vessel remodeling , and the ELISA method was used to detect levels of high density lipoprotein ( HDL-C) , cholesterol ( TC) and low density lipoprotein (LDL-C).The expressions of HMG-CoA reductase (HMGCR), low density lipoprotein receptor (LDLR) and ATP binding cassette transporter A1 (ABCA1) gene in the liver tissue were detected by real-time quantitative PCR.Results The right ventricular systolic pressure (40.12 ±8.22) mmHg(1 mmHg=0.133 kPa) and right ventricular hypertrophy index (0.352 ± 0.050) in the model group were significantly higher than those in the control group (P<0.05).The pulmonary artery vascular wall of the model group was significantly thicker than that of the control group .The HDL-C level of the model group was (26.20 ±3.73) mg/dl, which was significantly lower than that of the control group (P<0.05).There was no statistical significance in the levels of TC and

Objective To observe the expression of PAR2 and TMEM16A in the model of chronic constriction injury (CCI) in rat dorsal root ganglion (DRG) neurons,and to explore the role of it in the neuropathic pain.Methods Rats were divided into Sham operation group (Sham) and CCI group.Both groups were observed respectively to determine thermal withdrawal latency (TWL).The expression of PAR2 and TMEM16A in the dorsal root ganglion of the rat was analyzed using Western blot and immunofluorescence.Results The difference in preoperative TWL between CCI group and Sham group rats was not statistically significant (P ＜ 0.01).TWL was signifi cantly lower at all other time points after operation (P ＜ 0.01).Immunofluorescence results showed that PAR2 and TMEM16A coexisted in rat DRG neurons.Western blot results showed that,compared with Sham group,CCI group PAR2 and TMEM16A protein expression significantly increased after 7 d and 14 d (P ＜ 0.01),and the PAR2 and TMEM16A protein expression on 14 d is higher than that of 7 d (P ＜ 0.05).Conclusions Expression level of PAR2 and TMEM16A in CCI group was significantly higher than those in Sham group.The expression level of these proteins may be the cause of rat model of neuropathic pain.

Objective:To compare the therapeutic characteristics of anterior hybrid decompression and posterior cervical posterior laminectomy in the treatment of multilevel cervical spondylotic myelopathy.Methods:Thirty six cases of multilevel cervical spondylotic myelopathy patients treated by anterior hybrid decompression and thirty three cases of multilevel cervical spondylotic myelopathy patients treated by posterior cervical posterior laminectomy were involved.The general information,bleeding amount,operative time,cervical curvature D value,JOA score and incidence of postoperative complications of the two groups before and after surgery were compared.Results:There was no significant difference in the general information among the two groups(P＞0.05),including age (anterior group:56.23± 7.64 years old,posterior group:55.76± 8.18 years old),sex (anterior group:22 males/14 females,posterior group:20 males/13 females),cervical curvature D value (anterior group:7.41± 3.14,posterior group:8.19± 2.74),JOA score (anterior group:9.08± 1.09 scores,posterior group:8.82± 1.26 scores),disease course (anterior group:17.24± 7.36 months,posterior group:15.75± 5.78 months) and affected segment (anterior group:3.11 ± 0.26 segments,posterior group:3.24± 0.39 segments).The the amount of bleeding in the anterior group (anterior approach:221.79± 178.02 ml,posterior group:483.07± 434.25 ml) was lower than that of the posterior group(P＜0.05).The operative time (anterior group:196.54± 51.88 mins,posterior group:175.12± 54.93 mins) was longer,but there was no significant difference (P＞0.05).The cervical curvature D value and JOA score of posterior group were increased with the extension of surgery time.However,the cervical curvature D value of posterior group was decreased,but JOA score was increased.The incidence of bone unfinished,hoarseness and cerebrospinal fluid leakage were found in the anterior group,and axial pain and C5 nerve root paralysis were found in the posterior group.But there was no significant difference in the incidence of complications between the two groups (anterior group 14.89％,posterior group:12.12％)(P＞0.05).Conclusions:Anterior hybrid decompression and posterior cervical posterior laminectomy had their own advantages in the treatment of multilevel cervical spondylotic myelopathy.,The appropriate treatment should be taken according to the condition of patients.

Objective To investigate the effect of estrogen (E2) on the connexin43 (Cx43) expression of renal interlobar arteries after renal ischemia-reperfusion injury (I/R).Methods The experiment was carried out in vivo using an SD rat I/R model.SD rats were randomly divided into normal group,sham-operation group,I/R group,and estrogen-intervention group.The functional changes of the kidney were analyzed after 24 hours of I/R;nephridial tissue section was stained by hematoxylin-eosin (HE),and Paller scores were used to evaluate the degree of kidney damage.Pressure myography was utilized to detect the vasomotor function of renal interlobar arteries.Immunofluorescence technique,qRT-PCR and Western blot were applied to determine the expression of Cx43 in renal interlobar arteries in different groups.Results Estrogen markedly decreased the levels of Cr and BUN in the serum of I/R rats (P＜0.05),and the damage of the kidney tissue could be improved noticeably.The vasomotor rate of renal interlobar arteries was (24.80 ± 3.70)％ after I/R and (41.60 ± 3.50)％ after treatment with estrogen,which was higher than that of I/R group (P＜0.05).The expression of Cx43 was lower in renal interlobar arteries of estrogen-intervention group than that in I/R group (P＜0.01).Conclusion Estrogen may reduce vascular tension and boost dilation of the artery by inhibiting Cx43 expression and GJ function.Therefore,estrogen may attenuate the damage of I/R and improve renal function.

Objective To explore the relationship between KCNJ11-E23K polymorphism and essential hypertension (EH) in Kazakh from Xinjiang. Methods PCR-RFLP method was used to test KCNJ11-E23K genotypes of Kazakh from Xinjiang,including 237 EH patients and 221 normotension (NT). Logistic regression analysis was used to analyze the risk factors associated with EH. Results The frequencies of KCNJ11-E23K genotype (EE and (EK + KK)) and allele (E and K) were 34.18%, 65.82%, 61.60% and 38.40%respectively in EH group. There was a significant difference between two groups (P < 0.05). Weight and EE genotype were risk factors affecting EH in Kazakh from Xinjiang. Individual who carried EE genotype and allele E were 2.501 and 1.388 times than (EK + KK) and allele K suffered from EH respectively. Conclusion KCNJ11-E23K polymorphism was associated with EH in Kazakh from Xinjiang.

Objective To investigate the effects of propofol on mesenteric artery in SD rats and to observe whether the effect of propofol on the mesenteric artery relaxation is related to the gap. Methods Pressure myograph was used to examine the effect of 18β-GA and 2-APB on the relaxation induced by propofol 1×10 -7 ,3×10 -7 ,1×10 -6 ,3×10 -6 ,1 ×10 -5 ,3 ×10 -5 ,1 ×10 -4 and 3 ×10 -4 mol/L in acutely separated mesenteric arterioles of SD rat.Results The diameter of mesenteric arteri-oles were increased from (208.6±13.4)to (213.5±13.6),(21 9.7±13.2),(226.4±12.5),(234.9 ±12.3),(245.5±13.0),(267.4±1 5.2),(336.2±18.3)and (385.9 ±14.2)μm after application of 1×10 -7 ,3×10 -7 ,1 × 10 -6 ,3 × 10 -6 ,1 × 10 -5 ,3 × 10 -5 ,1 × 10 -4 and 3 × 10 -4 mol/L propofol,re-spectively.Propofol induced dilation of the rat mesenteric arterioles in a concentration-dependent man-ner (P < 0.01 ).After pretreatment with 18β-GA and 2-APB,1 × 10 -4 mol/L propofol induced dilation was absolutely decreased (P <0.01).Conclusion These results suggest that propofol relaxes mesenteric arterioles via gap junction.

Objective To investigate the effect of multidrug resistance-associated protein (MRP) 5 silenced by small interfering ribonucleic acid (siRNA) on the multi-drug resistance of human hepatocellular carcinoma (HCC) cells. Methods Multi-drug resistance human HCC cells HepG2/epirubicin(ADM) were constructed and MRP5-siRNA fragment was synthesized. The cells were transfected with Lipofectamine 2000. The experiment was divided into three groups. HepG2/ADM cells transfected with MRP5-siRNA were allocated in the siRNA group, HepG2/ADM cells in the drug resistance group and HCC cells HepG2 in the common group. The cellular sensitivity to chemotherapy agents was detected by MTT assay. The relative expression of MRP5 messenger ribonucleic acid (mRNA) was detected by real-time fluorescence quantiifcation RT-PCR. The expression of MRP5 protein was detected by Western blot. Cellular apoptosis was detected by flow cytometry. Multiple groups were compared by one-way ANOVA and two groups were compared by LSD-t test or t test. Results The half-inhibitory concentration (IC50) of cells for ADM, lfuorouracil (5-FU) and oxaliplatin (OXA) in the drug resistance group was (0.317±0.035), (3.785±0.523) and (0.129±0.009) mg/L, significantly higher compared with (0.022±0.008), (0.163±0.010) and (0.080±0.012) mg/L in the common group (t=14.202, 11.993, 13.937;P<0.05). The IC50 for the three chemotherapy drugs in the siRNA group was (0.180±0.008), (1.657±0.014) and (0.055±0.007) mg/L, signiifcantly lower than those in the drug resistance group (LSD-t=-6.609,-7.044,-11.257;P<0.05). The relative expression level of MRP5 mRNA in the drug resistance group was 3.858±0.481, signiifcantly higher compared with 1.000±0.374 in the common group (LSD-t=9.600, P<0.05). The relative expression level of MRP5 mRNA in the siRNA group was 1.377±0.141, signiifcantly lower than that in the drug resistance group (LSD-t=-11.669, P<0.05). The gray value of MRP5 protein was 2 245 in the drug resistance group, signiifcantly up-regulated compared with 58 in the common group. The gray value of MRP5 protein was 816 in the siRNA group, signiifcantly down-regulated compared with that in the drug resistance group. The cell apoptosis rate in the siRNA group was (25.1±3.7)%, significantly higher compared with (3.3±0.7)% in the drug resistance group (t=9.950, P<0.05). Conclusions MRP5 is associated with the multi-drug resistance of human HCC cells. MRP5 gene silenced by siRNA can enhance the sensitivity of HCC cells towards chemotherapy drugs.