Objective To investigate the effects of etomidate preconditioning on the expression of procaspase-3, caspase-9 p35 and caspase-8 p20 in HL-60 cells. Methods HL-60 cells were purchased from Shanghai life science institute and cultured in RPMI-1640 culture medium at 37℃ in 5% CO2 incubator. The cells were randomy divided into 3 groups ( n = 3 each): control group (group C), etomidate group (group E) and etomidate preconditioning group (group EP). In group E, the cells were exposed to 500 μmol/L etomidate and incubated for 24 h. In group EP, the cells were exposed to 1μmol/L etomidate for 1 h and was allowed to recover for4 h after etomidate washout, then etomidate 500μmol/L was added and the cells were incubated for 24 h. The expression of procaspase-3, caspase-9 p35 and caspase-8 p20 was determined using Western blot. Results The procaspase-3 expression was significantly down-regulated, while the expression of caspase-9 p35 and caspase-8 p20 was up-regulated ingroup E and EP as compared with group C ( P ＜ 0.05). The procaspase-3 expression was up-regulated,while the expression of caspase-9 p35 and caspase-8 p20 was down-regulated in group EP as compared with group E ( P ＜ 0.05). Conclusion Etomidate preconditiong can inhibit etomidate-induced down-regulation of procaspase3 expression and up-regulation of caspase-9 p35 and caspase-8 p20 expression, resulting in suppression of HL-60 cell apoptosis induced by etomidate.

Objective To evaluate the efficacy of flexible laryngeal mask airway and sevoflurane in pediatric strabismus surgery. Methods Totally 60 children undergoing elective strabismus surgery were recruited and divided into two groups with 30 cases each by random digits table:group T, in which children accepted endotracheal tube intubation and combined intravenous and inhalation anesthesia with mechanically ventilation; and group F, in which children adopted both flexible laryngeal mask airway and inhalation anesthesia of sevoflurane with spontaneous breath. Airway peak pressure (Ppeak), average airway pressure (Pmean), blood pressure (Bp) and heart rate (HR) at induction (T1), 1 min (T2), 3 min (T3) and 5 min (T4) after induction, removal of endotracheal tube or laryngeal mask airway (T5), awaking time and extubation time, occurrence of cough, dysphoria and pharyngodynia from being awake to extubation, satisfaction of patients were recorded. Results When compared to baseline, systolic pressure was lower at T, and higher at T2, T3 and T5, and HR was higher at T2-T5 in group T(P< 0.05),Bp was lower at T1-T4 in group F (P < 0.05). Bp and HR at T2-T5 in group F were lower than those in group T (P < 0.05). The awaking time was (8.1 ?2.6) min, the extubation time was (12.6 ?3.2) min, the occurrence of cough and pharyngodynia was 66.7%(20/30), 40.0%( 12/30), the dysphoria (0-1 score) was 7 cases in group T,(5.2 ± 1.1) min, (6.3 ± 1.5) rain,0,6.7%(2/30),25 cases in group F. There were significant differences between two groups (P<0.05). The satisfaction of patients in group F was higher than that in group T, there was significant difference between two groups (P< 0.05). Conclusion Flexible laryngeal mask airway with sevoflurane could offer high efficacy in pediatric strabismus surgery.

Objective To evaluate the effect of multiple-mode analgesia consists of epidural administration of lidocaine,parecoxib intravenously and patient-controlled epidural analgesia (PCEA) on patients after thoracic surgeries. Methods Total 90 patients treated with thoracic surgeries were allocated into 3 groups by random digits table,each with 30 cases. Patients accepted 1% lidocaine 5 ml epidurally 5 min before incision in group A and B,and 5 ml saline as control in group C. When chest closing parecoxib 40 mg were given in group A,and 2 ml saline as control in group B and C. All the patients were given PCEA. Postoperative hemodynamics,dosage of remifentanil and analgetics of PCEA within 48 h, visual analogue scale (VAS) of pain, rescue of pethidine, duration for first pain calling,and satisfactory scores of patients, doctors and nurses were recorded. Results Dosage of remifentanil was lower [(0.72±0.26), (0.84±1.20) μg／(kg·min)], and duration for first pain calling was longer[(35.00±2.78),(12.09±6.27) min] in group A and B than that in group C significantly[(1.80±0.84) μ,g／(kg·min) and (5.95±3.65) min](P<0.05). Analgetics of PCEA with in 48 h in group A was lower than that in group B which was lower than that in group C[(180.40±20.48), (192.54±40.15), (220.26±36.78) ml](P< 0.05). No difference was found on VAS of pain and hemodynamics among three groups when patients were kept immobilized, but VAS of pain in group A and B was lower than that in group C when patients kept moving. Satisfactory scores of patients,doctors and nurses in group A were higher. Conclusion Epidural administration of 1% lidocaine combined with intravenous paiecoxib 40 mg and PCEA an offer satisfactory postoperative analgesia for patients after elective thoracic surgeries.

Objective:Chemiluminescence immunoassay was used to detect the levels of anticardiolipin antibody (aCL) -IgA/IgG/IgM and anti-β2-glycoprotein Ⅰ antibody (aβ2GPⅠ) -IgA/IgG/IgM in healthy non-pregnant and pregnant women to explore the changes of antiphospholipid antibody in different pregnancy periods.Methods:This prospective study was conducted in Shandong Provincial Maternal and Child Health Care Hospital Affiliated to Qingdao University, involving normal pregnant women who underwent prenatal examination and healthy non-pregnant women with no history of adverse pregnancy who underwent progestational eugenic health examination from April 2020 to August 2021. The levels of aCL-IgA/IgG/IgM and aβ2GPⅠ-IgA/IgG/IgM were detected using BIO-FLASH chemiluminescence immunoassay analyzer and P95 as well as P99 were calculated, respectively. The difference in the six data between non-pregnant and pregnant women was compared using Mann-Whitney U test. Kruskal-Wallis H test was used to compare the change of each antibody in different pregnancy periods and Spearman correlation was used to analyze the correlation between different trimester and the levels of aCL-IgA/IgG/IgM and aβ2GPⅠ-IgA/IgG/IgM. Results:A total of 454 cases met the inclusion criteria, and 435 cases were included in the analysis after excluding 19 cases, among them 110 were non-pregnant women and 325 were pregnant women, including 110 cases in the first trimester (≤13 +6 weeks), 110 cases in the second trimester(14 +0-27 +6 weeks), and 105 cases in the third trimester (≥28 weeks). P99 value of aCL-IgA/IgG/IgM and aβ2GPⅠ-IgA/IgG/IgM in the non-pregnant women were 7.31, 14.70, 7.92, 3.58, 13.60, and 4.95 CU, which in the pregnant women were 5.90, 12.78, 5.70, 1.60, 10.65, and 3.90 CU, and were all lower than the cut-off value of 20 CU that given by the analyzer manufacturer. The levels of aCL-IgA/IgG/IgM, and aβ2GPⅠ-IgG/IgM in the pregnant women were significantly decreased comparing with the non-pregnant women [aCL-IgA: 1.90 CU (1.40-2.70 CU) vs 2.90 CU (2.20-3.83 CU), Z=－7.14; aCL-IgG: 3.00 CU (2.20-4.50 CU) vs 6.10 CU (4.20-7.83 CU), Z=－10.26; aCL-IgM: 1.40 CU (1.10-2.30 CU) vs 2.65 CU (2.08-3.73 CU), Z=－8.87; aβ2GPⅠ-IgG: 3.50 CU (2.60-4.90 CU) vs 4.75 CU (3.60-5.93 CU), Z=－5.45; aβ2GPⅠ-IgM: 0.70 CU (0.50-1.20 CU) vs 1.00 CU (0.60-1.53 CU) , Z=－3.73; all P<0.001]. The aCL-IgA level in the third trimester was higher than those in the first and second trimester (both P<0.05). The levels of aCL-IgG/IgM in the second trimester and aβ2GPⅠ- IgG in the second and third trimesters were significantly decreased than those in the first trimester (all P<0.05). Spearman analysis showed that aCL-IgG/IgM, aβ2GPⅠ-IgA/IgM had no significant correlation with the pregnancy period (the first, second and the third trimester) (all P>0.05). However, a weak correlation between the aCL-IgA, aβ2GPⅠ- IgG and the pregnancy period was observed ( r=0.28 and－0.49, both P<0.001) Conclusions:P99 value of aCL-IgA/IgG/IgM and aβ2GPⅠ-IgA/IgG/IgM levels in normal pregnant women and non-pregnant women are lower than the cut-off value of 20 CU given by the analyzer manufacturer. The levels of aCL-IgA/IgG/IgM and aβ2GPⅠ-IgG/IgM during pregnancy are lower than those before pregnancy and fluctuate with the pregnancy period, but have no significant correlation with the pregnancy period. The clinical diagnosis of antiphospholipid syndrome should be made according to the cut-off values of aCL-IgA/IgG/IgM and aβ2GPⅠ-IgA/IgG/IgM determined by each laboratory.

OBJECTIVETo evaluate the effect of quorum sensing luxS gene on biofilm formation through construction of a luxS overexpression strain by Streptococcus mutans (Sm).

METHODSIn order to construct pIB-luxS plasmid, the luxS gene fragment amplified by PCR was inserted into the shuttle plasmid pIB169 by corresponding double digests. The pIB-luxS plasmid was linearized electro-transformed into Sm cell and the overexpression strain was selected on chloramphenicol plate and testified by electrophoresis and western blot. The growth rate of both Sm wild type strain and its luxS overexpression strain were observed. Methyl thiazolyl tetrazolium (MTT) assay method was used to compare the biofilm formation quantification by both strains at different time points and containing different sucrose. The structures of the biofilms were observed by using confocal laser scanning microscopy, and biofilm-related gene expressions were investigated by real-time PCR. All experiments were performed in triplicate.

RESULTSThe luxS overexpression strain was successfully constructed and confirmed by electrophoresis and Western blotting. The planktonic growth mode of the wild-type and luxS overexpression strain showed no difference, but biofilm formed by Sm overexpression strain was 0.400 ± 0.009 and 0.609 ± 0.041 at 14 and 24 h, higher than the wild type strain biofilm at the same time point (0.352 ± 0.028 and 0.533 ± 0.014, respectively, P < 0.05). After adding 0.125% sucrose, biofilm formed by Sm overexpression strain raised to 1.041 ± 0.038, higher than that by the wild type strain (0.831 ± 0.020, P < 0.05). The biofilm formed by both strains were also increased with the sucrose concentration increase, but there was no difference between them. The overexpression strain aggregated into distinct clusters on structure, genes expression including gtfB, ftf, gbpB, relA, brpA, smu630, comDE, vicR were increased (6.10 ± 0.12, 3.34 ± 0.07, 8.75 ± 0.13, 2.96 ± 0.04, 5.20 ± 0.19, 2.20 ± 0.06, 2.32 ± 0.07 and 10.67 ± 0.57 fold) compared to the wild-type strain (P < 0.05).

CONCLUSIONSQuorum sensing luxS gene can promote the biofilm formation of Sm.

Bacterial Proteins ; genetics ; metabolism ; Biofilms ; growth & development ; Carbon-Sulfur Lyases ; genetics ; metabolism ; Microscopy, Confocal ; Plasmids ; genetics ; Quorum Sensing ; genetics ; Real-Time Polymerase Chain Reaction ; Streptococcus mutans ; physiology ; Tetrazolium Salts ; Time Factors

OBJECTIVETo investigate the predominant contribution of methyl-metabolism pathway to the regulation of LuxS of Strecptococcus mutans.

METHODSThe differences in biofilm formation and aciduricity of Strecptococcus mutans among the methyl-metabolism-complementation strain (KO-S), the parental wide-type strain (WT) and the luxS null strain (KO) were observed by real-time PCR for monitoring the transcriptional level of genes related to biofilm formation (smu.238, gtfD) and aciduricity (smu.44, smu.46) of the studied strains, methyl thiazolyl tetrazolium (MTT) for quantifying the biofilm of the exhibited strains and confocal laser scanning microscopy for estimating the structure of the biofilm.

RESULTSThe transcriptional level of smu.44, smu.46, smu.238, gtfD in WT were 1.289 ± 0.051, 1.694 ± 0.140, 1.565 ± 0.107, 1.667 ± 0.196 respectively; in KO were 1.001 ± 0.045, 1.007 ± 0.151, 1.000 ± 0.021, 1.012 ± 0.196 respectively, downregulated compared with WT (P < 0.05); in KO-S were 4.662 ± 0.091, 5.019 ± 0.258, 3.462±0.029, 3.071 ± 0.136 respectively, upregulated compared both with KO and with WT (P < 0.05). The quantity of biofilms formed by the studied strains were WT (1.592 ± 0.213), KO (0.939 ± 0.029), KO- S (2.177 ± 0.226), KO- P (1.020 ± 0.093), respectively, representing a less quantity by KO and KO-P than WT (P < 0.05) and a more quantity by KO-S than other three stains (P < 0.05). According to the observation of biofilms texture by confocal laser scanning microscopy, the WT biofilm was condensed and even. In contrast, fissures and gaps were found scattered in biofilms of KO, KO-P while lessened in that of KO-S, in which high-density bacterial aggregates were observed. The acid assay indicated a smaller biofilm decrease by WT and KO-S than that by KO and KO- P(P < 0.05).

CONCLUSIONSThe methyl- metabolism pathway contributes to LuxS regulation on biofilm formation and auiduricity of Strecptococcus mutans.

Bacterial Proteins ; metabolism ; Biofilms ; Carbon-Sulfur Lyases ; metabolism ; Glucosyltransferases ; Microscopy, Confocal ; Real-Time Polymerase Chain Reaction ; Streptococcus mutans ; metabolism

【Objective】 To analyze the changes of microRNA (miRNA) expression profiles on day 1 and day 5 after storage with or without riboflavin and ultraviolet-B (UVB) light (VB2-PRT) treatment, and to explore the molecular mechanism of miRNAs involved in the regulation of platelet storage lesion (PSL) under VB2-PRT treatment. 【Methods】 20 apheresis platelet concentrates (5mL / sample), collected from voluntary donors, were split into two group after mixing and agitation. One was treated with riboflavin (final concentration 50 μmol/L) plus 6.24 J/mL UVB light(E group), and the other worked as a control group (C group) without any treatment. Both groups were subjected to agitated storage at (22±2) ℃ horizontally. The platelet concentrates were sampled on d1 and d5 (5mL) during storage, named as E1, E5, C1 and C5 groups, respectively, and sequenced by DNA nanoball (DNB) sequencing technology. The differentially expressed miRNAs between E and C groups were screened by using DEGseq and MA-plot analysis software, and GO function enrichment analysis and KEGG pathway enrichment analysis were further performed when the different expression between groups reached twofold and above. 【Results】 Compared with C1 group, 487 miRNAs with significantly different expression (P<0.01) were screened in E1 group, including 220 up-regulated miRNAs, such as miR-146a and let-7b, and 267 down regulated miRNAs, such as miR-7 and miR-1260. Compared with C5 group, 229 miRNAs with significantly different expression (P<0.01) were screened in E5, including 80 miRNAs with up-regulated expression, such as miR-423 and miR-378, and 149 down regulated miRNAs, such as miR-451 and miR-30.The target genes with differentially expressed miRNAs in E1 vs C1 groups and E5 vs C5 groups were similar in the numbers of enriched GO terms, including cell components, organelles, cell membrane and other cell structures, molecular functions such as adhesion, catalysis, molecular transformation, transportation, transcription factors and receptor activity, cell processing, metabolism, biological regulation, stress and other biological processes etc. Compared with E1 and C1 groups, E5 and C5 groups lacked of signal pathways related to environmental adaptation, translation and mucin synthesis, however, it increased inositol phosphate metabolism, phosphatidylinositol signaling system and chemokine signaling pathway. 【Conclusion】 The expression profiles of platelets miRNAs treated with VB2-PRT has changed significantly after storage for a period of time. Functional prediction suggests that these miRNAs might be involved in the regulation of platelet PSL induced by VB2-PRT.