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.

Objective:To summarize the preliminary experience of endoscopically assisted mid-cheek benign tumor resection using a single preauricular or transoral incision and to evaluate its indications, advantages, and disadvantages.Methods:Thirty-six patients with benign mid-cheek tumors were prospectively enrolled, including 11 males and 25 females, aged (37.2± 15.9) years and ranged from 11 to 65 years old. The patients were randomly divided into two groups: endoscope-assisted tumor dissections through a single preauricular incision (preauricular group, 19 cases) or transoral incision (transoral group, 17 cases). Their surgical approaches were introduced, and the tumor long-axis length, incision length, operative time, estimated intraoperative bleeding, postoperative drainage amount and time, aesthetic satisfaction, perioperative complications, and follow-up were recorded and analyzed.Results:The difference between the tumor long-axis lengths in the preauricular group [(2.2±0.9) cm] and the transoral group [(2.1± 0.7) cm] was not statistically significant ( t=0.46, P=0.687), and all surgical procedures were completed as planned. There was no significant difference in the incision size ( t=1.57, P=0.100) or operative time ( t=0.44, P=0.736). Compared with the preauricular group [(30.8±8.7) ml], transoral group [(23.6±8.9) ml] significantly reduced intraoperative blood loss ( t=2.97, P=0.006) and improved aesthetic pleasure ( t=3.44, P=0.015). Two cases of earlobe numbness and one case of temporary facial palsy were observed in the preauricular group; two cases of postoperative effusion were noted in the transoral group, and no signs of nerve injury were detected. No tumor recurrence was found during the 1-54-month of follow-up. Conclusions:Endoscopic-assisted preauricular or transoral incision for dissecting mid-cheek benign tumors provides excellent aesthetic and minimally invasive results, reducing complications and obtaining satisfactory aesthetic results.

Objective: To investigate the role of connexin 43 ( Cx43) in acute sympathetic stress induced by phe- nylephrine (PE) overactivation of α1-adrenergic receptor ( α1-AR) in cardiomyocytes． Methods: Cardiomyocytes of H9C2 rats were randomly divided into control group，PE alone treatment group，Gap26 ( Cx43 specific inhibitor) intervention group and Gap26 alone treatment group．PE alone treatment group was treated with 50 μmol / L PE for 15 min．The Gap26 intervention group was pretreated with 0. 5 μmol / L Gap26 for 30 min，and then treated with 50 μmol / L PE for 15 min．The protein and mRNA expression levels of Cx43，NLRP3 inflammasome，interleukin-1 β (IL-1 β) ，Caspase-1，interleukin-18 (IL-18 ) were detected by Western blot and qRT-PCR. The expression and co-location of Cx43 in cardiomyocytes were observed by immunofluorescence assay，and the expression of inflamma- tory cytokines IL-1 β and IL-18 in cardiomyocytes was detected by ELISA. Results: Compared with control group， the protein and mRNA levels of Cx43，NLRP3，Caspase-1 and IL-18 in PE group increased．Compared with PE a- lone treatment group，the protein and mRNA levels of Cx43，NLRP3，Caspase-1 and IL-18 decreased after Gap26 intervention，but they were still higher than those of control group． Similarly ，immunofluorescence showed that Cx43 protein expression increased in PE alone group，while Cx43 expression was down-regulated in Gap26 inter- vention group compared with PE alone group．ELISA results showed that the expression of IL-1 β and IL-18 was sig- nificantly up-regulated in PE alone group，but down-regulated in Gap26 intervention group． Conclusion Cx43 is involved in α1-AR activation induced cardiac acute sympathetic stress by regulating NLRP3 inflammasome.

【Objective】 To study the mechanism of Apocynum leaves in the treatment of sepsis by network pharmacology method in order to explore the multi-dimensional research method of Xinjiang ethnic medicine treatment of infectious diseases and provide scientific theoretical basis for clinical medication. 【Methods】 TCMSP and literature collection were used to screen the main active components of Apocynum venetum leaf, and target prediction analysis was conducted by SwissTarget Prediction database. We used Genecards database to screen relevant targets for sepsis, used Omicshare to calculate Venn figure of the intersection targets, constructed the protein-protein interaction network diagram in the STRING database, used Ensembl for name conversion of protein targets, then entered Omicshare server for Go function and KEGG pathway enrichment of dynamic analysis. Last we used Cytoscape3.6.0 software to construct "Apocynum venetum leaf-the active ingredients-targets-Go-KEGG-sepsis" network to explore the mechanism of action of Apocynum’s resistance to sepsis. 【Results】 Kaferol, luteolin, proanthocyanidin B1 and phytosterol in Apocynum venetum leaves may treat sepsis by controlling signal transduction, regulating hormone level and anti-infection through predictive biological targets such as Akt1, VEGFA, MAPK3, EGFR, Src and PTGS2. They can play an important role through VEGF, EGFR, erbB, HIF-1, RAS and other enrichment pathways. 【Conclusion】 The active components of Apocynum venetum leaves can fight against sepsis through multiple targets and multiple pathways.

Objective:To evaluate the relationship between transforming growth factor beta-3 (TGF-β 3)/mammalian homologs of the drosophila mad gene 3 (Smad3) signaling pathway and neuronal apoptosis during reduction of focal cerebral ischemia-reperfusion (I/R) injury by isoflurane postconditioning (ISO) in rats. Methods:Sixty clean-grade healthy male Sprague-Dawley rats, aged 6-8 weeks, weighing 210-230 g, were randomly divided into 5 groups ( n=12 each): sham operation group (group S), cerebral I/R group (group I/R), cerebral I/R plus isoflurane postconditioning group (group I/R+ ISO), cerebral I/R plus pirfenidone group (group I/R+ P), and cerebral I/R plus isoflurane postconditioning plus pirfenidone group (group I/R+ ISO+ P). Local cerebral I/R was produced by middle cerebral artery occlusion for 1.5 h followed by 24-h reperfusion in anesthetized rats.Pirfenidone 5 μg/kg was injected into the lateral ventricle at 30 min before ischemia in I/R+ P and I/R+ ISO+ P groups.In I/R+ ISO and I/R+ ISO+ P groups, 1.5% isoflurane was inhaled for 1 h starting from the time point immediately after onset of reperfusion.Neuro-functional deficit was assessed using neurologic deficit scores (NDS) at the end of reperfusion.Then the animals were sacrificed, and brain tissues were removed for determination of the neuronal damage rate (by Nissl staining), neuronal apoptosis rate (by TUNEL), expression of TGF-β 3 (using immunofluorescence), and expression of TGF-β 3, phosphorylated Smad3 (p-Smad3), caspase-3, Bax and Bcl-2 (by Western blot). Results:Compared with group S, the NDS, neuronal damage rate and apoptosis rate of neurons were significantly increased, the expression of TGF-β 3, caspase-3 and Bax was up-regulated, and the expression of p-Smad3 and Bcl-2 was down-regulated in group I/R ( P<0.05). Compared with group I/R, the NDS, neuronal damage rate and apoptosis rate of neurons were significantly decreased, the expression of TGF-β 3, p-Smad3 and Bcl-2 was up-regulated, and the expression of caspase-3 and Bax was down-regulated in group I/R+ ISO, and the NDS, neuronal damage rate and apoptosis rate of neurons were significantly increased, the expression of TGF-β 3 and p-Smad3 was down-regulated, and the expression of caspase-3 was up-regulated in group I/R+ P ( P<0.05). Compared with group I/R+ ISO, the NDS, neuronal damage rate and apoptosis rate of neurons were significantly increased, the expression of TGF-β 3, p-Smad3 and Bcl-2 was down-regulated, and the expression of caspase-3 and Bax was up-regulated in group I/R+ ISO+ P ( P<0.05). Conclusion:Isoflurane postconditioning can inhibit neuronal apoptosis by activating the TGF-β 3/Smad3 signaling pathway, thus reducing focal I/R injury in rats.

Objective: To introduce a new surgical procedure for the treatment of neck benign tumors by endoscopic techniques. Methods: Seventeen patients with neck benign tumor underwent surgery by endoscope through a concealed incision in Department of Oral and Maxillofacial Surgery, Qilu Hospital of Shandong University from January 2018 to August 2019 were analyzed, which included 3 cases of tumor in the submental area, 2 cases in submandibular region, 9 cases in lower pole region of parotid gland, 1 case in superior region of sternocleidomastoid muscle, 1 case in central region of sternocleidomastoid muscle, 1 case in inferior region of sternocleidomastoid muscle. All patients underwent routine preoperative examination and CT examination to evaluate tumor size, boundary, morphology and nature. According to the area where the tumor located, concealed incisions in different sites were designed. Lumps in the submental area and submandibular area were treated with oral vestibular sulcus incision. Benign tumors located in the lower pole region of parotid gland and the sternocleidomastoid muscle region were treated with approach of the short hidden postauricular incision. During the operation, the self-developed "maxillofacial suspension device" was used to provide the operating space. The tumors were completely removed with endoscope and all patients were followed up every 3 months. Results: All surgical procedures were performed as expected. Visual analogue scale (VAS) was 9.3 on average at 3 months after operation, all the patients were satisfied with the incision design and the cosmetic effect. No recurrences were found in patients with a follow-up period ranged from 1-15 months. Conclusions These studies have shown that endoscope-assisted neck benign tumor resection is a surgical procedure with covert incision and good cosmetic results.

Objective: To summarize the preliminary experience of endoscope -assisted resection of superficial parotid gland benign tumors, and to discuss the indications, advantages and disadvantages of the operation. Methods: The clinical data of 18 patients who underwent extracapsular resection of superficial parotid gland benign tumor in Department of Oral and Maxillofacial Surgery, Qilu Hospital of Shandong University from March 2018 to March 2019 were retrospectively analyzed, and the surgical methods were introduced. The indications, long axis length of tumor, incision design, operation time, intraoperative blood loss, postoperative drainage and drainage time, aesthetic satisfaction, postoperative complications and follow-up time were counted. Results: All procedures were completed as expected. The length of the long axis of the tumor was (2.3±0.6) cm, the incision in the tragus around the earlobe was short and concealed, the incision length was (5.1±1.3) cm, the operation duration was (2.0±0.4) h, intraoperative blood loss was (168.9±18.8) ml, postoperative drainage was (29.5±11.7) ml, drainage time was (3.6±0.5) d, 2 cases of temporary facial paralysis or earlobe numbness, three months after the operation, the results of the visual analogue scale of the incision design and the aesthetic effect were (9.6±0.1). Conclusions Endoscope-assisted resection of superficial parotid gland benign tumor by inner tragus around earlobe approach is applicable and reliable, can reduce complications, shorten surgical incision to obtain satisfactory aesthetic effect, which is worth further expansion and improvement.

Objective To study the changes of intracellular calcium ion concentration in pulmonary artery smooth muscle cells (PASMCs) of hypoxic-induced persistent pulmonary hypertension (PPH) induced by calcium-sensitive receptor (CaSR) in a newborn mouse model.Method Ninety-six newbom C57BL/6 mice were randomly divided into control group,PPH group,PPH + agonist group and PPH + inhibitor group,with 24 mice in each group.The PPH model was induced by 12％ oxygen for 14 days.In the beginning,intraperitoneal injection of CaSR agonist (GdCl3) and CaSR inhibitor (NPS2390) were performed to mice in PPH + agonist group and PPH + inhibitor group respectively daily.After 14 days of modeling,pulmonary artery smooth muscle cells (PASMCs) of all four groups were cultured in vitro.Changes of Ca2+ fluorescence intensity in PASMCs of the four groups were detected by laser confocal microscope continuously.Result The ratio of pulmonary small vascular wall thickness to the vascular diameter and right ventricle/left ventricular thickness in PPH group were greater than those in the control group [(21.1％ ±1.8％) vs.(27.0％ ±0.9％),(0.62 ±0.22) vs.(0.83±0.45)],the differences were statistically significant (P ＜ 0.05),which imply that PPH mouse model was constructed successfully.The average Ca2+ fluorescence intensity in PASMCs of control group,PPH group,PPH + agonist group and PPH+ antagonist group were 122.5 ± 3.0,2 058.8 ±46.3,2 286.6 ±51.4 and 1 134.8 ± 8.5,respectively.The average Ca2+ fluorescence intensity in PASMCs of the PPH group,PPH + agonist group and PPH + antagonist group was higher than that of the control group respectively,the average Ca2+ fluorescence intensity in PASMCs of PPH group was higher than that of PPH + antagonist group,the differences were statistically significant (P ＜ 0.05).Whereas the difference of average Ca2 + fluorescence intensity in PASMCs of PPH group and PPH + agonist group was of no statistical significance (P ＞ 0.05).Conclusion CaSR may be involved in the occurrence and development of hypoxic-induced PPH in neonatal mice by affecting the intracellular Ca2+ concentration in pulmonary artery smooth muscle cells.

Objective To evaluate the effects of remifentanil (RMF)on large conductance cal-cium-activated potassium channel (BKCa)and voltage-gated potassium channel (KV)activition currents in basilar arterial smooth muscle cells (BASMCs)of normotensive and hypertensive rats. Methods Spontaneously hypertensive rats (SHR)and homologous normotensive wistar-kyoto (WKY)rats,were used in this study.BASMCs were obtained freshly by the method of enzymolysis. Six basilar artery smooth muscle cells of each rat were chosen and analyzed.Outward current ampli-tude was recorded by the whole-cell patch clamp technique.The outward current amplitude under all stimulation voltage in set of step stimulation protocol before (basal level)and after administration of RMF (3×10-7mol/L)were recorded respectively and net current was calculated (net current=cur-rent amplitude after administration-basic value).With administration by concentrations cumulative method,the current amplitude under +60 mV stimulation voltage was separately recorded before (basic value)and after application of different concentrations of RMF (10-10,10-9,10-8,10-7, 10-6,10-5mol/L),then calculated current increasing rate and the half effective concentration (EC50)of RMF increasing current amplitude in BASMCs.Another six basilar artery smooth muscle cells of each rat were chosen and given RMF (3×10-7mol/L),and separately treated with BKCa channel blocker (tetraethylammonium,TEA)and Kv channel blocker (4-aminopyridine,4-AP),and then administrated the corresponding RMF mixture.The current amplitude was recorded after each dose.Results At 0,+20,+40 and +60 mV,the net current generated by RMF on both BASMCs of rats was successively and significantly increased (P <0.05).The increment rate of outward currents in BASMCs generated by 10-10,10-9,10-8,10-7RMF successively and significantly went upward (P<0.05).Compared to WKY rats,the half-effective concentration(EC50)of RMF increas-ing the current amplitude in BASMCs of SHR significantly rose(P<0.05).Compared with the base-line,the current amplitude in BASMCs of the two kind rats was significantly increased after adminis-tration of RMF,and decreased after administration of TEA or 4-AP (P<0.05);Compared to ad-ministration of TEA or 4-AP,the current amplitude in BASMCs of the two kind rats was significantly in-creased after administration of TEA+RMF or 4-AP+RMF (P<0.05).Conclusion Bkcaand Kv currents in both BASMCs of SHR and WKY rats were activated by RMF in a voltage-dependent and dose-dependent manner,and the effect of RMF on BKCaand Kvactivition currents in BASMCs of SHR was weaker than WKY rats.

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.