SARS-CoV-2 spike protein receptor binding domain promotes IL-6 and IL-8 release via ATP/P2Y2 and ERK1/2 signaling pathways in human bronchial epithelia.

The spike protein of SARS-CoV-2 as well as its receptor binding domain (RBD) has been demonstrated to be capable of activating the release of pro-inflammatory mediators in endothelial cells and immune cells such as monocytes. However, the effects of spike protein or its RBD on airway epithelial cells and mechanisms underlying these effects have not been adequately characterized. Here, we show that the RBD of spike protein alone can induce bronchial epithelial inflammation in a manner of ATP/P2Y2 dependence. Incubation of human bronchial epithelia with RBD induced IL-6 and IL-8 release, which could be inhibited by antibody. The incubation of RBD also up-regulated the expression of inflammatory indicators such as ho-1 and mkp-1. Furthermore, ATP secretion was observed after RBD treatment, P2Y2 receptor knock down by siRNA significantly suppressed the IL-6 and IL-8 release evoked by RBD. Additionally, S-RBD elevated the phosphorylation level of ERK1/2, and the effect that PD98059 can inhibit the pro-inflammatory cytokine release suggested the participation of ERK1/2. These novel findings provide new evidence of SARS-CoV-2 on airway inflammation and introduce purinergic signaling as promising treatment target.

Compact continuous-wave solid-state Raman lasers at 707 and 714†nm for laser trapping and cooling of atomic strontium and radium.

Two compact laser sources at 707 and 714 nm are realized efficiently by using a diode-pumped a-cut Nd:YVO4 laser with intracavity stimulated Raman scattering and sum-frequency generation (SFG). The fundamental wave at 1342 nm is generated by the 4F3/2 → 4I13/2 transition in Nd:YVO4 crystal. The Raman Stokes waves at 1496 and 1526 nm were obtained by placing the c-axis of the Nd:YVO4 crystal along the Ng and Nm axes of an Np-cut KGW crystal, respectively. LBO crystals with critical phase matching are used to perform the intracavity SFG of fundamental and Stokes waves. At a pump power of 36 W, the maximum output powers at 707 and 714 nm can reach 2.72 and 3.14 W, corresponding to light-to-light conversion efficiencies of 7.5% and 8.7%, respectively. The developed 707 and 714 nm laser sources are practically useful in laser trapping and cooling related to atomic strontium and radium.

Selectable two-wavelength Nd:YVO4 Raman laser at 671 and 714†nm.

A compact efficient continuous wave (CW) laser with selectable two wavelengths at 671 and 714 nm is developed. The laser cavity comprises an Nd-doped and an undoped YVO4 crystal to generate the fundamental wave at 1342 nm and the first-Stokes Raman wave at 1525 nm, respectively. A single LBO crystal with the cut angle in the XZ plane is designed to achieve the selectable phase-matching via the thermal tuning for the second harmonic generation (SHG) of 1342 nm and the sum frequency generation (SFG) of 1342 and 1525 nm. At a pump power of 40 W, the optimal output powers at 671 and 714 nm can reach 4.5 and 1.8 W, respectively. The present compact CW laser source at 671 and 714 nm has practical usefulness for laser spectroscopy and numerous applications.

ß2-Adrenoceptor Activation Stimulates IL-6 Production via PKA, ERK1/2, Src, and Beta-Arrestin2 Signaling Pathways in Human Bronchial Epithelia.

OBJECTIVE: ß2-Adrenoceptor agonists are widely used to treat asthma because of their bronchial-dilation effects. We previously reported that isoprenaline, via the apical and basolateral ß2-adrenoceptor, induced Cl- secretion by activating cyclic AMP (cAMP)-dependent pathways in human bronchial epithelia. Despite these results, whether and how the ß2-adrenoceptor-mediated cAMP-dependent pathway contributes to pro-inflammatory cytokine release in human bronchial epithelia remains poorly understood. METHODS: We investigated ß2-adrenoceptor-mediated signaling pathways involved in the production of two pro-inflammatory cytokines, interleukin (IL)-6 and IL-8, in 16HBE14o- human bronchial epithelia. The effects of isoprenaline or formoterol were assessed in the presence of protein kinase A (PKA), exchange protein directly activated by cAMP (EPAC), Src, and extracellular signal-regulated protein kinase (ERK)1/2 inhibitors. The involvement of ß-arrestin2 was examined using siRNA knockdown. RESULTS: Isoprenaline and formoterol (both ß2 agonists) induced IL-6, but not IL-8, release, which could be inhibited by ICI 118,551 (ß2 antagonist). The PKA-specific inhibitor, H89, partially inhibited IL-6 release. Another intracellular cAMP receptor, EPAC, was not involved in IL-6 release. Isoprenaline-mediated IL-6 secretion was attenuated by dasatinib, a Src inhibitor, and PD98059, an ERK1/2 inhibitor. Isoprenaline treatment also led to ERK1/2 phosphorylation. In addition, knockdown of ß-arrestin2 by siRNA specifically suppressed cytokine release when a high concentration of isoprenaline (1 mM) was used. CONCLUSION: Our results suggest that activation of the ß2-adrenoceptor in 16HBE14o- cells stimulated the PKA/Src/ERK1/2 and/or ß-arrestin2 signaling pathways, leading to IL-6 release. Therefore, our data reveal that ß2-adrenoceptor signaling plays a role in the immune regulation of human airway epithelia.

Evaluation of a novel Cardiac Peri-Operative Transfusion Trigger Scoring system in patients with coronary artery disease.

BACKGROUND: A simple and accurate scoring system to guide perioperative blood transfusion in patients with coronary artery disease (CAD) undergoing cardiac surgery is lacking. The trigger point for blood transfusions for these patients may be different from existing transfusion guidelines. This study aimed to evaluate the safety and efficacy of a new scoring strategy for use in guiding transfusion decisions in patients with CAD. METHODS: A multicenter randomized controlled trial was conducted at three third-level grade-A hospitals from January 2015 to May 2018. Data of 254 patients in a Cardiac Peri-Operative Transfusion Trigger Score (cPOTTS) group and 246 patients in a group receiving conventional evaluation of the need for transfusion (conventional group) were analysed. The requirements for transfusion and the per capita consumption of red blood cells (RBCs) were compared between groups. RESULTS: Baseline characteristics of the two groups were comparable. Logistic regression analyses revealed no significant differences between the two groups in primary outcomes (1-year mortality and perioperative ischemic cardiac events), secondary outcomes (shock, infections, and renal impairment), ICU admission, and ICU stay duration. However, patients in the cPOTTS group had significantly shorter hospital stays, lower hospital costs, lower utilization rate and lower per capita consumption of transfused RBCs than controls. Stratified analyses revealed no significant differences between groups in associations between baseline characteristics and perioperative ischemic cardiac events, except for hemofiltration or dialysis and NYHA class in I. CONCLUSIONS: This novel scoring system offered a practical and straightforward guideline of perioperative blood transfusion in patients with CAD. Trial registration chiCTR1800016561(2017/7/19).

Ginsenoside Rg1 protects rat bone marrow mesenchymal stem cells against ischemia induced apoptosis through miR-494-3p and ROCK-1.

This study aimed to verify the cytoprotective effect of ginsenoside Rg1 in vivo, and to elucidate the mechanism of Rg1 in the ischemic microenvironment. Male rat bone marrow mesenchymal stem cells (rBMSCs) or rBMSCs treated with Rg1 were injected into ischemic region of the arterial embolism hind limb in female rats. Behavioral and histological data, obtained one-week post injection, showed that rBMSCs with Rg1 could improve the survival rate of BMSCs and enhance the therapeutic effects. rBMSCs treated with hypoxia and serum deprivation for 24h (H/SD-rBMSCs) showed the up-regulated expression of ras homolog family member A (RhoA), Rho associated coiled-coil containing protein kinase 1 (ROCK-1), myosin light chain 2 (MLC-2), Bcl2 associated agonist of cell death (Bad) and Bcl2 associated X, apoptosis regulator (Bax); while the expression of miR-148b-3p, miR-148b-5p and miR-494-3p was down-regulated. H/SD with Rg1 treatment (H/SD+Rg1-rBMSCs) inhibited the expression of ROCK-1, MLC-2, Bad and Bax, increased the expression of Bcl-2, miR-494-3p. After ROCK-1 knockout, the expression of Bad and Bax were downregulated and Bcl-2 upregulated, but Rg1 no longer altered their expression. Mir-494-3p functional study established that miR-494-3 mimic downregulated and miR-494-3 inhibitor upregulated ROCK-1 gene expression, Rg1 did not have the ability to change the ROCK gene expression after loss of function of miR-494-3p. Also, the function loss of mir-494-3p promoted apoptosis; otherwise reduced apoptosis. The anti-apoptotic effect of Rg1 disappeared after mir-494-3p loss or gain function. In conclusion, Ginsenoside Rg1 has shown to have protective effects on ischemic-induced rBMSCs apoptosis through mir-494-3p→ROCK-1→Bcl-2 signaling pathway.

The influence of rapamycin on the early cardioprotective effect of hypoxic preconditioning on cardiomyocytes.

INTRODUCTION: The purpose of this study was to examine the effects of rapamycin on the cardioprotective effect of hypoxic preconditioning (HPC) and on the mammalian target of rapamycin (mTOR)-mediated hypoxia-inducible factor 1 (HIF-1) signaling pathway. MATERIAL AND METHODS: Primary cardiomyocytes were isolated from rat pups and underwent rapamycin and/or HPC, followed by hypoxia/re-oxygenation (H/R) injury. Cell viability and cell injury were determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and lactate dehydrogenase (LDH) assays, and qRT-PCR was used to measure HIF-1α and mTOR mRNA expression. A Langendorff heart perfusion model was conducted to observe the effect of rapamycin. RESULTS: Rapamycin treatment nearly abolished the cardioprotective effect of HPC in cardiomyocytes, reduced cell viability (p = 0.007) and increased cell damage (p = 0.032). HIF-1α and mTOR mRNA expression increased in cardiomyocytes undergoing I/R injury within 2 h after HPC. After rapamycin treatment, mTOR mRNA expression and HPC-induced HIF-1α mRNA expression were both reduced (p < 0.001). A Langendorff heart perfusion model in rat hearts showed that rapamycin greatly attenuated the cardioprotective effect of HPC in terms of heart rate, LVDP, and dp/dtmax (all, p < 0.029). CONCLUSIONS: Rapamycin, through inhibition of mTOR, reduces the elevated HIF-1α expression at an early stage of HPC, and attenuates the early cardioprotective effect of HPC.

Carcinoma of the Bartholin Gland: A Review of 33 Cases.

OBJECTIVE: Primary carcinoma of the Bartholin gland is a rare malignancy that accounts for approximately 5% of vulvar carcinomas. The aim of the study was to compare the outcomes of women with primary Bartholin gland carcinoma (BGC) with those with non-Bartholin gland-related vulvar carcinoma. MATERIALS AND METHODS: A retrospective chart review of 429 patients with invasive vulvar carcinoma evaluated at a single institution between 1993 and 2011 was performed. Medical records were reviewed for demographic data, pathologic information, treatment type, and recurrence/outcome information. These variables were compared between patients with primary BGC and patients with non-Bartholin gland-related vulvar carcinoma. RESULTS: Thirty-three (7.7%) of the 429 patients with invasive vulvar carcinoma had primary carcinoma of the Bartholin gland. Twenty-nine patients (87.9%) had squamous cell histology and 4 patients (12.1%) had adenocarcinoma. When compared with non-Bartholin gland-related vulvar carcinoma, patients with primary BGC had a younger age at diagnosis (median, 57 vs 63 years; P = 0.045), had a higher rate of stage III/IV disease (60.6% vs 35.8%; P = 0.008), and were more likely to receive radiation therapy (78.8% vs 43.9%; P < 0.001). However, there were no significant differences between the 2 groups with regard to histologic subtype, lymphovascular space involvement, perineural invasion, positive margins, recurrence-free survival, or overall survival. CONCLUSIONS: Despite being diagnosed at a more advanced stage, patients with primary carcinoma of the Bartholin gland seem to have similar oncologic outcomes and survival rates to patients with non-Bartholin gland-related vulvar carcinoma.

ePIANNO: ePIgenomics ANNOtation tool.

Recently, with the development of next generation sequencing (NGS), the combination of chromatin immunoprecipitation (ChIP) and NGS, namely ChIP-seq, has become a powerful technique to capture potential genomic binding sites of regulatory factors, histone modifications and chromatin accessible regions. For most researchers, additional information including genomic variations on the TF binding site, allele frequency of variation between different populations, variation associated disease, and other neighbour TF binding sites are essential to generate a proper hypothesis or a meaningful conclusion. Many ChIP-seq datasets had been deposited on the public domain to help researchers make new discoveries. However, researches are often intimidated by the complexity of data structure and largeness of data volume. Such information would be more useful if they could be combined or downloaded with ChIP-seq data. To meet such demands, we built a webtool: ePIgenomic ANNOtation tool (ePIANNO, http://epianno.stat.sinica.edu.tw/index.html). ePIANNO is a web server that combines SNP information of populations (1000 Genomes Project) and gene-disease association information of GWAS (NHGRI) with ChIP-seq (hmChIP, ENCODE, and ROADMAP epigenomics) data. ePIANNO has a user-friendly website interface allowing researchers to explore, navigate, and extract data quickly. We use two examples to demonstrate how users could use functions of ePIANNO webserver to explore useful information about TF related genomic variants. Users could use our query functions to search target regions, transcription factors, or annotations. ePIANNO may help users to generate hypothesis or explore potential biological functions for their studies.

The cardioprotective effect of hypoxic and ischemic preconditioning in dogs with myocardial ischemia-reperfusion injury using a double-bypass model.

AIMS: The effects of preconditioning on cardioprotection have mainly been studied in vitro. No sufficient in vivo experiments have been performed to optimize ischemic preconditioning (IPC) or hypoxic preconditioning (HPC) for clinical applications. The purpose of this study was to establish a canine double-bypass model to examine the effect of IPC and HPC on cardiomyocytes and heart function. MATERIALS AND METHODS: A double-bypass procedure to enable independent control of systemic and coronary circulation was established in dogs. The animals were divided into control, HPC, and IPC groups (n=6 each). Indicators of cardiac function, including cardiodynamics, hemodynamics, ATP, and cardiac troponin I (cTnI) levels; myocardium morphology; and myocardiocyte apoptosis were determined. KEY FINDINGS: Both IPC and HPC attenuated the reperfusion-induced decrease in left ventricular end systolic pressure seen in the control group. Both the HPC and IPC groups had lower serum cTnI levels, better myocardiocyte histology, and lower rates of apoptosis compared to the control group without preconditioning. HPC reduced the abnormal cardiomyocyte histology and apoptosis to a greater extent than IPC, and only HPC significantly restored the depletion of ATP. SIGNIFICANCE: This study demonstrates the effectiveness of the double-bypass model for the optimized study of both HPC and IPC. The results suggest that HPC may provide better cardioprotection than IPC.

Anti-apoptotic role of peroxiredoxin III in cervical cancer cells.

As a member of peroxiredoxin (Prx) family, PrxIII is predominantly located in mitochondria and plays an important role as a scavenger of reactive oxygen species (ROS). Since previous reports demonstrated over-expression of PrxIII in cervical cancer, we conducted the present study to investigate the significance of PrxIII in cervical cancer development and/or progression. Cervical cancer cells were cultured from tissues derived from cervical cancer patients. After successful knockdown of PrxIII expression by small interfering RNA, we evaluated ROS level, viable cell number, and apoptosis of cervical cancer cells along with the culture time. The production of ROS was increased in cervical cancer cells as compared with normal cervical epithelia. Knockdown of PrxIII expression induced up-regulation of other Prx members including PrxI, PrxII, and PrxV. ROS level was higher in down-regulated cervical cancer cells than in controls and the difference was increasing with culture time. We also observed increased apoptosis and decreased viable cell number in down-regulated cervical cancer cells. Our results suggest that PrxIII is an indispensable ROS scavenger, which protects tumor cells against oxidative damage and subsequent apoptosis.

[Comparison of the cardioprotection between crystalloid and blood cardioplegia in adult patients undergoing cardiac surgery: a meta-analysis].

OBJECTIVES: To compare the cardioprotection effect between blood and crystalloid cardioplegia during cardiac surgery in adult patients, and provide a theoretical basis for optimal myocardial protection strategies. METHODS: A meta-analysis of randomized controlled trials (RCT) studies about comparing blood and crystalloid cardioplegia in adult patients undergoing cardiac surgery were performed. Cochrane library (Issue 3, 2011), MEDLINE, EMBase, PubMed, HighWire, CBM and CNKI were searched from January 1985 to December 2011. Studies were assessed according to the Cochrane Handbook for systematic reviews. Data were extracted from these trials and analyzed by RevMan5.1 software. RESULTS: Sixteen trials involved 3934 patients were included, 2004 cases were in blood group, and 1930 were in crystalloid group. There was no statistical heterogeneity between studies using a fixed effects model. Meta-analysis indicated that, there were no significant differences between blood and crystalloid group in the incidence of postoperative 30 days mortality (OR = 1.11, 95%CI: 0.59 - 2.08, P = 0.74), the incidence of postoperative low cardiac output (OR = 0.98, 95%CI: 0.41 - 2.33, P = 0.85), the incidence of perioperative myocardial infarctions (OR = 0.85, 95%CI: 0.55 - 1.29, P = 0.44), and inotropic support requirement (OR = 1.05, 95%CI: 0.81 - 1.38, P = 0.70). CONCLUSION: The blood cardioplegia is no difference with crystalloid cardioplegia in adult patients undergoing cardiac surgery.

Sevoflurane at 1 MAC provides optimal myocardial protection during off-pump CABG.

OBJECTIVE: We investigated the myocardial protective effect of sevoflurane in patients receiving off-pump coronary artery bypass grafting (OPCABG) and the role of brain natriuretic peptide (BNP). DESIGN: Forty-eight patients receiving elective OPCABG were randomly assigned to a control group, and to 0.75 MAC, 1.0 MAC and 1.5 MAC sevoflurane groups. Blood samples were collected and levels of BNP and cardiac troponin I (cTnI) were measured before anesthesia, and immediately, 24, 48 and 72 h after surgery. RESULTS: Dopamine was necessary to maintain blood pressure in the sevoflurane groups, but not in the control group (p < 0.002). 1.0 MAC sevoflurane significantly decreased post-surgical cTnI levels (p < 0.001). 0.75 MAC had no significant effect, and increasing sevoflurane concentrations to 1.5 MAC caused no further decrease in cTnI concentrations. There was no significant difference in BNP level among the groups (p = 0.227) or between any two groups, although values of BNP showed a significant correlation with cTnI values in control subjects immediately after (r = 0.847) and 24 h after (r = 0.661) surgery. CONCLUSIONS: Our results demonstrated that 1.0 MAC and 1.5 MAC sevoflurane can exert a significant myocardial protective effect. BNP cannot be used to predict the myocardial protective effect of sevoflurane in OPCABG.

[Effect of perioperative glucose-insulin-potassium infusions on the prognosis in patients undergoing coronary artery bypass grafting: a meta-analysis].

OBJECTIVE: To assess the effect of perioperative glucose-insulin-potassium (GIK) infusions on the prognosis in patients undergoing coronary artery bypass grafting. METHODS: Electronic databases including Cochrane library (Issue 3, 2011), Pubmed, EMbase, Highwire, CBM and CNKI were searched. A meta-analysis of all randomized controlled trials (RCTs) comparing GIK with control in coronary artery bypass grafting was performed. Study selection and meta-analysis were conducted which according to the Cochrane Handbook for systematic reviews. Date were extracted from these trials by 3 reviewers independently and analyzed by RevMan5.0 software. RESULTS: A total of 9 RCTs including 1029 patients were assessed in this study. GIK infusion was associated with significantly fewer perioperative myocardial infarctions (RR = 0.59, 95%CI: 0.38 - 0.91, P = 0.02), less inotropic support requirement (RR = 0.44, 95%CI: 0.35 - 0.56, P < 0.01), and increase the incidence of postoperative atrial fibrillation (RR = 1.23, 95%CI: 1.05 - 1.43, P = 0.009). CONCLUSIONS: GIK significantly reduces myocardial injury and improves cardiac function in patients undergoing coronary artery bypass grafting, but also increases the incidence of postoperative atrial fibrillation.

Synthesis, crystal structures, and biological evaluation of Cu(II) and Zn(II) complexes of 2-benzoylpyridine Schiff bases derived from S-methyl- and S-phenyldithiocarbazates.

Two NNS tridentate Schiff base ligands of 2-benzoylpyridine S-methyldithiocarbazate (HL(1)) and 2-benzoylpyridine S-phenyldithiocarbazate (HL(2)) and their transition metal complexes [Cu(2)(L(1))(2)(CH(3)COO)](ClO(4)) (1), [Zn(2)(L(1))(2)(ClO(4))(2)] (2), [Zn(L(2))(2)](3) have been prepared and characterized by elemental analysis, IR, MS, NMR and single-crystal X-ray diffraction studies. In the solid state, each of two Schiff bases remains in its thione tautomeric form with the thione sulfur atom trans to the azomethine nitrogen atom. Under similar prepared conditions, three new complexes showed distinctly different coordination modes depending on their coordinating preferences. Each copper atom in S-bridged dinuclear complex [Cu(2)(L(1))(2)(CH(3)COO)](ClO(4)) (1) is surrounded by five donor atoms in a square-pyramidal fashion (4+1). [Zn(2)(L(1))(2)(ClO(4))(2)] (2) is a dimer in which each zinc atom adopts a seven-coordinate distorted pentagonal bipyramidal geometry, while mononuclear [Zn(L(2))(2)] (3) has octahedral coordination geometry. Biological studies, carried out in vitro against selected bacteria, fungi, and K562 leukaemia cell line, respectively, have shown that different substituted groups attached at the dithiocarbazate moieties and metals showed distinctive differences in the biological property. Zinc(II) complexes 2 and 3 could distinguish K562 leukaemia cell line from normal hepatocyte QSG7701 cell line. Effect of the title compounds on Mitochondria membrane potential (MMP) and PI-associated fluorescence intensity in K562 leukaemia cell line are also studied. The title compounds may exert their cytotoxicity activity via induced loss of MMP.

Mn(II), Co(II) and Zn(II) complexes with heterocyclic substituted thiosemicarbazones: synthesis, characterization, X-ray crystal structures and antitumor comparison.

Transition metal complexes Mn(L(1))(2) (1), Mn(L(2))(2) (2), Co(L(3))(2)Cl 4H(2)O (3), Zn(L(3))(2) DMF (4), Co(HL(4))(2)(ClO(4))(2) 3H(2)O (5) and Zn(L(5))(2) DMF (6) where HL(1)=2-acetylpyridine thiosemicarbazone, HL(2)=2-acetylpyridine N(4)-methylthiosemicarbazone, HL(3)=2-benzoylpyridine thiosemicarbazone, HL(4)=2-benzoylpyridine N(4)-methylthiosemicarbazone and HL(5)=2-benzoylpyridine N(4)-phenylthiosemicarbazone, have been synthesized. The complexes 1, 2, 5 and 6 were characterized by elemental analysis, IR spectra and single-crystal X-ray diffraction studies. Preliminary in vitro screening indicated that all the tested compounds showed significant antitumor activity against K562 leucocythemia cancer cell line.

[Effect of MAPK signal transduction pathway inhibitor U0126 on aquaporin 4 expression in alveolar type II cells in rats with oleic acid-induced acute lung injury].

OBJECTIVE: To investigate the relationship between aquaporin 4 (AQP4) in alveolar type II (AT-II) cells and MAPK signaling pathway in rats with early-stage oleic acid-induced acute lung injury (ALI) and acute respiratory distress syndrome (ARDS). METHODS: Three groups of rats, namely the normal control, ALI and U0126 treatment group were used in this study. After oleic acid-induced ALI in the latter two groups, the rats in the treatment group received 100 micromol/L U0126 treatment at the dose of 10 micro, and dimethyl sulfoxide (DMSO) were given in the normal control and ALI groups. Arterial blood gas and the extravascular lung water (EVLW) content were measured after the treatments, and pathological changes in the lung tissues were observed microscopically. ATII cells were isolated from the lung tissues and identified using tannic acid staining and alkaline phosphatase (APK) staining. The expression of AQP-4 mRNA in the cells was detected with RT-PCR. RESULTS: Blood gas analysis, HE staining and EVLW content measurement revealed severer injury of the lung tissues in ALI group than in the normal control group, but the severity was comparable between the treatment and ALI groups. RT-PCR demonstrated significantly increased AQP-4 mRNA expression in ALI group as compared with that in the normal control group, and U0126 treatment resulted in obvious reduction in AQP-4 mRNA expression in the U0126 treatment group. CONCLUSION: Oleic acid-induced ALI results in the activation of MAPK signaling pathway and up-regulation of AQP-4 mRNA expression in the ATII cells of rats.

[Changes of the sodium channel in alveolar type II cells in pulmonary water transport in rats with oleic acid-induced acute lung injury].

OBJECTIVE: To determine the capability of alveolar fluid clearance and the changes of sodium channel in alveolar type II cells (ATII) in oleic acid-induced acute lung injury. METHODS: Forty four male Sprague-Dawley (SD) rats were randomized into a control group and an acute lung injury (ALI) group, with 22 rats in each group. The ALI model was established by oleic acid. The ATII cells were acutely isolated and purified, and the ATII cellular ultrastructure was observed by transmission electron microscope. In each group, the mRNA expression of 3 epithelial sodium channel (ENaC) subunits in acute isolated ATII cells from 8 rats were detected by reverse transcription-polymerase chain reaction (RT-PCR), while the extravascular lung water (EVLW) content was quantified in 7 rats by gravimetric measurement, and the lung histopathological changes were studies in 7 rats. RESULTS: In the ALI group, Smith lung injury score (7.6 +/- 0.8) and EVLW (0.80 +/- 0.17) ml were significantly higher than those in the control group [Smith score: (1.1 +/- 0.2), t = -20.859, P < 0.01; EVLW: (0.52 +/- 0.10) ml, t = -3.851, P < 0.01]. The transmission electron microscopic observation showed that there were degeneration, apoptosis, and lamellar body vacuolar changes in the ATII cells from the ALI rats. RT-PCR demonstrated that the alpha-subunit of the ENaC mRNA expression was the highest among the 3 subunits (F = 4.40, P = 0.02). In the ALI group, mRNA expressions of all the 3 ENaC subunits in acutely isolated ATII cells were decreased as compared to those in the control group [alpha-subunit: (51 +/- 9)% vs (82 +/- 7)%, t = 7.61, P < 0.01; beta-subunit: (13 +/- 7)% vs (25 +/- 4)%, t = 4.53, P < 0.01; gamma-subunit: (31 +/- 15)% vs (40 +/- 17)%, t = 3.01, P < 0.05; respectively]. CONCLUSIONS: The capacity of alveolar fluid clearance was attenuated in oleic acid-induced acute lung injury. The ENaC subunit mRNA levels of ATII cells were significantly decreased in ALI rats.

[Expression of Notch intracellular domain in cervical cancer and effect of DAPT on cervical cancer cell].

OBJECTIVE: To study the expression and clinical significance of Notch intracellular domain (NICD) in cervical cancer and the effects of N-[N-(3,5-difluorophenyl)acetyl-L-alanyl]-S-phenyl glycine t-butyl ester (DAPT), a gamma-secretase inhibitor on the proliferation and apoptosis of cervical cancer cell lines. METHODS: Western blot was used to detect the expression of NICD in the tissues of 40 cervical cancers and 21 normal cervix and its relationship with clinical features of cervical cancer was also analyzed. Proliferation of SiHa and HeLa cervical cells was determined by methyl thiazolyl tetrazolium (MTT) assay, cell cycles and apoptosis and index of proliferation were detected by flow cytometry method. The expression of NICD in SiHa and HeLa cells incubated with DAPT was detected by western blot. RESULTS: The expression level of NICD in cervical cancers was significantly higher than that of normal cervical tissues (1.237 +/- 0.353 vs 0.938 +/- 0.105, P < 0.05). The NICD expression was higher in cervical cancers with high grade, lymph node involvement and parametrial invasion than that with low-middle grade (1.496 +/- 0.540 vs 1.150 +/- 0.216), without lymph node involvement (1.419 +/- 0.532 vs 1.159 +/- 0.210) and no parametrial invasion (1.718 +/- 0.710 vs 1.183 +/- 0.258), respectively (all P < 0.05). The expression of NICD in cervical adenocarcinoma was higher than that of squamous cell cancer (1.463 +/- 0.395 vs 1.162 +/- 0.187, P < 0.05). After SiHa and HeLa cells were incubated with DAPT, NICD expression was significantly lower than that in control (P < 0.05). The effects of DAPT inhibited the proliferation and prompted the apoptosis of SiHa and HeLa cells was depended on its concentrations and times. CONCLUSIONS: NICD may play a key role in the occurrence and progress of cervical cancer. The mechanism of DAPT inhibited the proliferation and prompted the apoptosis of SiHa and HeLa cells may be due to decreased the formation of NICD.

Cytotoxicity and structure-activity relationships of four alpha-N-heterocyclic thiosemicarbazone derivatives crystal structure of 2-acetylpyrazine thiosemicarbazone.

A series of thiosemicarbazone ligands, HL(1) (2-acetylpyrazine thiosemicarbazone), HL(2) (2-acetylpyrazine N(4)-methylthiosemicarbazone), HL(3) (2-benzoylpyridine thiosemicarbazone) and HL(4) (2-benzoylpyridine N(4)-methylthiosemicarbazone), have been synthesized. The crystal structure of HL(1) has been determined by single-crystal X-ray diffraction. Hydrogen bonds link the different components to stabilize the crystal structure. The antitumor activity of the four ligands were tested against K562 leucocythemia and BEL7402 liver cancer cell lines. All the thiosemicarbazones showed significant antitumor activity. Different substituents on the ligands show different levels of antitumor activity. By comparison with the other thiosemicarbazone species studied, HL(4) with substitution at N(4) position in thiosemicarbazone along with 2-benzoylpyridine is the most active thiosemicarbazone ligand with IC(50)=0.002microm in the K562 leucocythemia cell line and 0.138microm in the BEL7402 liver cancer cell line, respectively.