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.

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).

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.

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.

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.

Molecular mechanism of ALKBH5-mediated m6A demethylation regulating lipopolysaccharide-induced epithelial-mesenchymal transition in sepsis-induced acute kidney injury.

This study explored the mechanism by which the m6A demethylase ALKBH5 mediates epithelial-mesenchymal transition (EMT) in sepsis-associated acute kidney injury (SA-AKI) and AKI-chronic kidney disease (CKD) transition. HK-2 cells were stimulated with lipopolysaccharide (LPS) to establish an in vitro model of SA-AKI. ALKBH5 expression was reduced through the transfection of si-ALKBH5. Cell viability, apoptosis, and migration were detected by CCK-8 assay, TUNEL staining, and Transwell. The levels of TNF-α, IL-1ß, and IL-6 were measured by enzyme-linked immunosorbent assay. Quantitative real-time polymerase chain reaction or Western blotting was performed to determine the expressions of ALKBH5, miR-205-5p, DDX5, E-cadherin, and α-SMA. The m6A level was quantitatively analyzed. The expression of pri-miR-205 bound to DGCR8 and m6A-modified pri-miR-205 after intervention with ALKBH5 expression was detected by RNA immunoprecipitation. A dual-luciferase assay confirmed the binding between miR-205-5p and DDX5. ALKBH5 was highly expressed in LPS-induced HK-2 cells. Inhibition of ALKBH5 increased cell viability, repressed apoptosis, and reduced EMT. Inhibition of ALKBH5 increased the m6A modification level, thereby promoting DGCR8 binding to pri-miR-205 to increase miR-205-5p expression and eventually targeting DDX5 expression. Low expression of miR-205-5p or overexpression of DDX5 partially abolished the inhibitory effect of ALKBH5 silencing on EMT. In conclusion, ALKBH5 represses miR-205-5p expression by removing m6A modification to upregulate DDX5 expression, thereby promoting EMT and AKI-CKD transition after SA-AKI.

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.

[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.

[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.

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.

1,2-Bis(1H-pyrrol-2-ylmethyl-ene)diazane monohydrate.

The mol-ecular structure of title compound, C(10)H(10)N(4)·H(2)O, has an inversion centre located on the mid-point of the N-N bond of the mol-ecule. A twofold rotation axis passes through the water O atom. In the crystal structure, a two-dimensional network is constructed through N-H⋯O and O-H⋯N hydrogen bonds.

Bis(2,2'-bipyridine-κN,N')(thio-cyanato-κN)copper(II) perchlorate.

The asymmetric unit of title compound, [Cu(NCS)(C(10)H(8)N(2))(2)]ClO(4), contains a bis-(2,2'-bipyridine)(isothio-cyanato)copper(II) cation and a perchlorate anion. In the cation, the Cu(2+) ion is coordinated by four N atoms from two bidentate 2,2'-bipyridine mol-ecules and an N atom from an isothio-cyanate anion, resulting in a distorted CuN(5) pyramidal configuration. The crystal structure is stabilized by weak inter-molecular C-H⋯O and C-H⋯S hydrogen bonds, and weak π-π inter-actions between 2,2'-bipyridine rings [centroid-centroid distance = 3.908 (4) Å]. The perchlorate counteranion is disordered over two positions in a 0.66:0.34 ratio.

[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.

[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.

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.

N'-Ferrocenyl-2-hydroxy-benzohydrazide.

The title complex, [Fe(C(5)H(5))(C(13)H(11)N(2)O(3))], was prepared via self-assembly using ferrocenyl hydrazide and ethyl salicylate. The compound is potentially a tridentate ferrocene-based ligand. The conformation of the mol-ecule allows the formation of an intra-molecular N-H⋯O hydrogen bond involving the hydroxyl group. The CONHNHCO unit and the rings bonded to it are nearly coplanar. The crystal structure is stabilized by inter-molecular O-H⋯O(carbon-yl) and N-H⋯O(carbon-yl) hydrogen bonds.

2-Acetyl-pyrazine 4-methyl-thio-semi-carbazone.

The title compound, C(8)H(11)N(5)S, has been prepared by the reaction of 2-acetyl-pyrazine with 4-methyl-3-thio-semi-carbazide. It exists in the thione form and adopts the E configuration. The mol-ecules are connected by the inter-molecular N-H⋯N and N-H⋯S inter-actions.

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.

[Functional study of progesterone receptor isoforms in endometrial cancer cell lines].

OBJECTIVE: To study the functional differences between the two progestrone receptor isoforms(PR-A and PR-B) in human endometrial cancer,using antisense oligodeoxynucleotide(AS-ODN) to downregulate isoform B of progestrone receptor in endometrial carcinoma cell lines, After transfection of the oligodeoxynucleotide, several kinds of hormones were added in the cells to observe the different response, where to study the functional differences between the two isoforms. METHODS: The well-differentiated endometrial cancer cell line Ishikawa and moderate-differentiated endometrial cancer cell line Hec-1B were cultured in vitro. The cells were transfected with antisense, sense, and scramble-ODN. After 48 hours, the expressions of two progesterone receptor isoforms were detected by Western blot using specific antibody. Then the cells were planted in 96-well plates, transfected with antisense, sense, scramble-ODN and added in several hormones to search for the response in distinct hormones and oligodeoxynucleotides. RESULTS: After transfecting antisense-ODN, two cell lines were down-regulated in progesterone receptor isoform B,but progesterone receptor isoform A was not down-regulated,and the progesterone receptor isoform B of cells transfected with sense and scramble-ODN was not changed. When stimulated by 17beta-estradiol(E2)for 72 hours,the growth of Ishikawa cells was significantly higher than that of the control, Hec-1B cells only grew higher than control,but it was to significant in statistics.R5020 inhibited Ishikawa cells significantly after stimulating for 72 hours. There was the same effect in Hec-1B cells after stimulating for 96 hours. On the bases of E2 and R5020, we added mifepristone (RU486) . The cells developed after 96 hours in Ishikawa cells and developed after 48 hours in Hec-1B cells. When PR-B was down-regulated,the stimulating effect of E2 was enhanced, but the inhibitory effect of R5020 was decreased, RU486 antagonized R5020 weaklier than the control. CONCLUSION: AS-ODN directed against the human PR-B can inhibit the expression of PR-B effectively,through which the PR-A expresses predominantly. E2 can cause endometrial carcinoma cell growth, PR-B is associated with the stimulating effect of E2 in endometrial carcinoma cells. Progestin (R5020) inhibits the hyperplasia induced by E2,PR-B is involved in the inhibitory effect of R5020. RU486 antagonizes the effect of R5020,inhibiting cell growth, PR-B is involved in the antagonizing effect of RU486.