[Efficacy and safety of combination therapy with chemotherapy, programmed death-1 inhibitor and anlotinib in the treatment of advanced dedifferentiated liposarcoma].

Objective: To investigate the efficacy and safety of the combination therapy with chemotherapy, programmed death-1 (PD-1) inhibitor and anlotinib in the treatment of advanced dedifferentiated liposarcoma (DDLPS). Methods: The clinical data of patients with dedifferentiated liposarcoma who received chemotherapy combined with PD-1 inhibitor and anlotinib in the Department of Medical Oncology, Zhongshan Hospital Affiliated to Fudan University from January 1, 2020 to November 30, 2021 were retrospectively analyzed. A total of 24 patients were included in this study, including 12 males and 12 females, with a median age of onset of 56 years (range, 31-69 years). Efficacy and safety in those patients were assessed. Results: All patients had unresectable or metastatic dedifferentiated liposarcoma with G2 (moderate differentiation) or G3 (differential differentiation) in a concise three-grade grading scheme of tumor pathology. Twelve patients received the regimen as the first-line treatment, while the other 7 taken the regimen as second-line treatment and 5 as third-line or above. The median follow-up time for overall survival (OS) was 7.7 months. The overall response rate (ORR) was 20.8% (5/24) and disease control rate (DCR) was 83.3% (20/24) with 5 partial response (PR), 15 stable disease (SD) and 4 progressive disease (PD). Overall, the median progression-free survival (PFS) was 4.9 months (95%CI: 3.4-16.2 months). The ORR of anthracycline-based, eribulin-based or gemcitabine-based regimens was 1/12, 2/6 and 2/6, respectively; and the median PFS was 7.7, 7.3 and 4.4 months, respectively. Waterfall plots showed notable tumor shrinkage of any degree in eribulin and gemcitabine-based regimens(3/6 and 2/6, respectively), while there were more patients presented with SD in anthracycline-based group(9/12). Common adverse reactions included myelosuppression, fatigue, anorexia, rash, pruritus, palpitate, hypothyroidism and hypertension. Conclusions: The combination regimen with chemotherapy, PD-1 inhibitor and anlotinib in the treatment of advanced DDLPS is effective and well tolerable. There are more responders in eribulin or gemcitabine-based regimens.

Minocycline relieves myocardial ischemia-reperfusion injury in rats by inhibiting inflammation, oxidative stress and apoptosis.

OBJECTIVE: Myocardial ischemia-reperfusion (I/R) injury (MIRI) is an important cause of irreversible injury to the myocardium in patients with acute myocardial infarction. The purpose of this study was to investigate the effects of minocycline (MC) on inflammation, oxidative stress and apoptosis of myocardial tissues. MATERIALS AND METHODS: We used rats to establish MIRI model by ligating coronary arteries. The structure and function of rat myocardium were determined by 2, 3, 5-triphenyl tetrazolium chloride (TTC) staining, hematoxylin-eosin (HE) staining and echocardiography. In addition, we detected the expression of inflammatory factors, antioxidant enzymes and apoptosis-related molecules in rats by enzyme-linked immunosorbent assay (ELISA), immunohistochemical (IHC) staining and reverse transcription-polymerase chain reaction (RT-PCR) to determine the effect of MC on inflammation, oxidative stress and apoptosis in I/R rats. Finally, we studied the effect of MC stimulation on the viability of rat cardiomyocytes (H9c2 cells) in vitro. RESULTS: After I/R, the heart function of rats decreased, and the structure of myocardium was destroyed. The levels of inflammation and oxidative stress in I/R rats also increased significantly, manifested by increased inflammatory factors and decreased antioxidant enzymes in serum and myocardial tissue. After treatment of I/R rats with MC, the structure and function of rat myocardium improved significantly, and MC reduced inflammation and oxidative stress levels in rats, thus inhibiting the apoptosis of cardiomyocytes. MC also improved the viability of H9c2 cells in vitro. CONCLUSIONS: MC reduced inflammation and oxidative stress levels in MIRI rat model or H9c2 cells, thus inhibiting cardiomyocyte apoptosis. Therefore, MC has potential application prospects for the treatment of MIRI.

[Adenocarcinomas with mesonephric features in gynecologic tract: a clinicopathological analysis of seven cases].

Objective: To investigate and compare the histologic characteristics of adenocarcinomas with mesonephric features located in different parts of the gynecologic tract. Methods: Two cases of mesonephric adenocarcinomas (MA) of the cervix and 5 cases of mesonephric-like adenocarcinomas (MLA) of the uterus and ovary were collected in Women's Hospital, School of Medicine, Zhejiang University from January 2018 to October 2020. Hematoxylin-eosin staining, immunohistochemistry and KRAS mutation testing were performed together with review of literature. Results: MA of the cervix as well as MLA of the uterus and ovary had similar morphologic features, showing an admixture of glandular, tubular, papillary and solid growth patterns. However, both MA cases were located in cervical stroma, which demonstrated residual mesonephric ducts present at the periphery. All four uterine MLA cases extensively involved the endometrium and myometrium. The ovarian MLA case was associated with endometriosis. No residual mesonephric ducts were present in the MLA cases. Immunohistochemically, GATA3 was positive in all seven MA/MLA cases. TTF1 was expressed only in 4/5 MLA cases. ER and PR were negative and p53 was wild-type in all cases. KRAS mutation was detected in all five cases. During the 6-32 months of follow-up, one patient developed recurrence and the others were tumor-free. Conclusions: In the gynecologic tract, both MA in cervix and MLA in uterus and ovary have similar morphologic features, immunohistochemical expression and KRAS mutation. However, distinct from MA that originates from mesonephric remnant, MLA is closely related to Mullerian epithelium.

[Comparison of extracorporeal membrane oxygenation applicated in critical patients with COVID-19 and novel influenza A (H1N1) virus pneumonia].

Objective: To compare the clinical characteristics and prognosis of critical patients with COVID-19 and novel influenza A (H1N1) virus pneumonia (influenza pneumonia) applied with extracorporeal membrane oxygenation (ECMO). Methods: A total of 24 patients with influenza pneumonia treated with ECMO in respiratory intensive-care unit (ICU) of Beijing Chaoyang Hospital from March 2016 to December 2019 and 12 patients with COVID-19 hospitalized from February 1, 2020 to March 31, 2020 in 5 government designated infectious hospitals of Beijing and Hebei Province that applied with ECMO were enrolled. The demographic data, clinical manifestations, and ECMO related information were described and analyzed and all numerical variables are described as M (P25, P75). Results: The age of COVID-19 patients was 77 (66, 79) years old, which was older than influenza pneumonia patients [46 (32, 62) years old], P<0.05; acute lung injury score and respiratory ECMO survival prediction (RESP) score before ECMO application were 3.3 (3.0, 3.5) and 1 (0, 2), respectively, which were lower than influenza pneumonia patients [3.8 (3.5, 4.0) and 4 (2, 6), respectively], all P values<0.05. Thrombotic complications, bleeding complications, and ventilator-associated pneumonia occurred in ECMO applied COVID-19 patients were 4, 10 and 5 cases, respectively, which were more than that among influenza pneumonia patients (1, 9, and 2 cases, respectively), all P values<0.05. The length of ICU stay of COVID-19 patients was 31 (28, 75) d, which was longer than that of influenza pneumonia patients [27 (18, 39) d], P<0.05. The cases with successful decannulation of ECMO among COVID-19 and influenza pneumonia patients were 6 and 14 cases, respectively and mortality during ICU stay were 8 cases and 11 cases, respectively, and the difference were not statistically significant (all P values>0.05). Conclusion: COVID-19 patients applied with ECMO have more ECMO-related complications and a longer stay in the ICU than patients with influenza pneumonia.

LncRNA AFAP1-AS1 promotes proliferation ability and invasiveness of bladder cancer cells.

OBJECTIVE: It was the aim of this study to explore the role and mechanism of long non-coding RNA (lncRNA) AFAP1-AS1 in the progression of bladder cancer (BCa) by in vitro experiments. PATIENTS AND METHODS: AFAP1-AS1 levels in 40 pairs of clinical BCa tissue samples and normal ones collected from BCa patients were determined, and paired sample t-test was applied to compare the differences between groups. The prognosis data of patients with BCa were collected, and survival analysis and t-test were performed to specify the interplay between AFAP1-AS1 and the prognosis of BCa patients. Subsequently, AFAP1-AS1 expression level in BCa and normal cells were further confirmed by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR), and Cell Counting Kit-8 (CCK-8), 5-Ethynyl-2'-deoxyuridine (EdU), and transwell assays were performed to figure out the influence of this lncRNA on the proliferation ability and invasiveness of BCa cells. Meanwhile, the interaction between AFAP1-AS1 and its sense mRNA was analyzed. We used co-transfection technology to simultaneously transfect si-AFAP1-AS1 and pcDNA3.1-AFAP1 or their corresponding negative controls into BCa cells, and cell proliferation and invasion ability in different subgroups were determined to explore the underlying mechanism through which AFAP1-AS1 plays a role in BCa progression. RESULTS: No matter in BCa tissues or in cell samples, compared to the corresponding normal controls, AFAP1-AS1 was found highly expressed; at the same time, in invasive bladder cancer tissues, the expression level of AFAP1-AS1 was also higher than that in non-invasive tissues. Meanwhile, survival analysis revealed that patients with BCa with high expression of AFAP1-AS1 owned a shorter overall survival rate than those with low expression, indicating a negative interplay between AFAP1-AS1 expression and patients' prognosis. In addition, in BCa cell lines, according to the results of CCK-8, EDU, and transwell assays, the proliferative capacity, as well as the invasive ability of BCa cells, were found weakened after downregulation of AFAP1-AS1. Meanwhile, a negative interplay was discovered between AFAP1-AS1 and its sense mRNA. Finally, the results of cell reversal experiment using co-transfection technique revealed that overexpression of AFAP1 can reverse the inhibitory impact of lncRNAAFAP1-AS1 on the malignant ability of BCa cells. CONCLUSIONS: AFAP1-AS1 may enhance the proliferation ability as well as the invasiveness of BCa cells so as to aggravate the degree of BCa malignancy.

[Analysis of special ehealth service for corona virus disease 2019 (COVID-19) pneumonia].

OBJECTIVE: To analyze how governments, hospitals and information technology(IT) companies use Internet technology to provide online health services during the early stage of corona virus disease 2019 (COVID-19) epidemic in January 2020 in China, and then provide suggestions and coping strategies for the later stage and post-epidemic time. METHODS: We searched for information on ehealth services related to the outbreak of COVID-19 in China. The sources of information were mainstream search engines such as Baidu and the popular interactive social platforms such as Webchat. The keywords were "Internet+pneumonia", "Internet clinic", "pneumonia online clinic" and so on. The time of information was from January 20 to February 3, 2020. The key information was extracted and encoded by two persons back-to-back. The coding information included: name of organization provider, launching time, location of provider, service items, user, health workers engaging in the service, and so on. The coded information was entered and analyzed with SPSS 24.0 and Excel. RESULTS: There were totally 57 projects launched by local governments, hospitals and IT companies. Most of them were launched from January 24th to 27th, the hospital and government projects services regionally, especially in eastern provinces. In this study, 90.48% of the enterprises and 100.00% of the hospitals had online fever clinic and consultation services for COVID-19, 66.67% of the enterprises and 37.04% of the hospitals serviced derivative health problems. Only a few projects provided tele-medical consultation. There were individual projects that provided online health management for home quarantine people. Physicians were the main force of various projects. In some hospital projects, there were also nurses, pharmacists and professional technicians to provide featured consultation. CONCLUSION: Ehealth is useful and helpful for the health care system to rapidly cope with health demand during instantaneous and post epidemic time. Regional distribution of ehealth is unbalanced. There are institutional and technical feasibilities for the emergency application of Internet technology. However, community health centers seldom provide ehealth or connect with tertiary hospitals with Internet. Therefore, all kinds of providers within healthcare system should promote emergence ehealth. Tele-medical diagnosis and referral should be developed by local governments during COVID-19. The application of "Internet+medical treatment" in community medical institutions and synergy among various institutions should be promoted.

Upregulation of lncRNA DANCR functions as an oncogenic role in non-small lung cancer by regulating miR-214-5p/CIZ1 axis.

OBJECTIVE: Lung cancer has an unfavorable prognosis due to the lack of efficient diagnostic and therapeutic strategies. Therefore, this study sought to figure out the effect of long non-coding RNA (lncRNA) DANCR on lung cancer progression. PATIENTS AND METHODS: LncRNA DANCR and miR-214-5p expressions in non-small cell lung cancer (NSCLC) were detected by Real Time-quantitative Polymerase Chain Reaction (RT-qPCR). Function assays, including Cell Counting Kit-8 (CCK-8) and flow cytometric analysis were conducted to clarify the role of DANCR and miR-214-5p in the progression of NSCLC. Western blot, Dual-Luciferase reporter assay, and RNA immunoprecipitation assay (RIP) were performed to elucidate the underlying mechanism. RESULTS: LncRNA DANCR was upregulated in NSCLC. The knockdown of lncRNA DANCR inhibited cell proliferation and accelerated cell apoptosis in NSCLC. LncRNA DANCR interacted with miR-214-5p. MiR-214-5p over-expression partially reversed the regulatory effects of DANCR on proliferation and apoptosis in NSCLC. In addition, CIZ1 was the downstream gene binding miR-214-5p. LncRNA DANCR could regulate the miR-214-5p/CIZ1 axis. CONCLUSIONS: Downregulation of lncRNA DANCR inhibited cell proliferation and induced cell apoptosis in NSCLC by regulating the miR-214-5p/CIZ1 axis. LncRNA DANCR may act as an oncogene and promote the progression of NSCLC.

Notch signaling is involved in regulation of LPS-induced macrophage apoptosis through JNK/NF-kB signaling pathway.

Notch signaling plays a critical role in the development and function of macrophages. The aim of the present study was to investigate the relationship between Notch signaling pathway and macrophage apoptosis after LPS stimulation. In RAW 264.7 cells, the mRNA expression of Jagged1, Hes1, Hes 5 and GM-CSF, and protein expression of NICD1 and GM-CSF were increased after LPS stimulation. Inhibition of Notch signaling by γ-secretase inhibitor DAPT and the suppression of Notch1 expression using siRNA both significantly prevented LPS induced activation of JNK and NF-kB, and simultaneously the expression of GM-CSF was also down regulated significantly. JNK inhibitor SP600125 was used to block the phosphorylation of JNK signaling, Western blot results showed that the activation of NF-kB was blocked and expression of GM-CSF was down-regulated. Finally, flow cytometry analyses showed that the Notch signaling was involved in the regulation of macrophage apoptosis after LPS stimulation. Our study showed that the Notch signaling pathway was activated and involved in the regulation of macrophage apoptosis after LPS stimulation through JNK/ NF-kB signaling regulated GM-CSF expression.

[Short-term outcomes of minimally invasive reoperation for tricuspid regurgitation after left-sided valve surgery].

Objective: To examine the short-term outcomes of minimally invasive reoperation for severe tricuspid regurgitation after left-sided valve surgery. Methods: From January 2015 to December 2018, a total of 89 patients with severe tricuspid regurgitation after left-sided valve surgery received reoperation in Department of Cardiovascular Surgery, Zhongshan Hospital, Fudan University were included in this study. There were 21 males and 68 females, aging of (56.4±7.9) years (range: 41 to 74 years). The interval between previous left-sided valve surgery and tricuspid reoperation was (14.1±6.1) years (range: 4 to 33 years). A combination of multiple minimally invasive techniques were adopted, including endoscopy-assist right minithoracotomy approach, peripheral cannulation strategy with the vacuum-assist single venous drainage technique, heart beating technique, and temporary percutaneous pacemaker implantation, with a concomitant enhancement in preoperative right cardiac function optimization. Results: All patients received minimally invasive isolated tricuspid valve replacement (n=81) or tricuspid valve repair (n=8). After the application of multiple minimally invasive techniques, the operative mortality rate was only 3.4% (3/89). The causes of death were progressive right heart failure with multiorgan failure (n=1) and low cardiac output associated with postoperative bleeding (n=2). Regarding to the perioperative complications, renal replacement therapy rate was 5.6% (5/89), permanent pacemaker implantation rate was 1.1% (1/89), and the incidence of stroke was 0. Mechanical ventilation time was 24(24) hours, ICU stay time was 2.5 (3.0) days (M(Q(R))). During the short-term follow-up, there were no case of severe tricuspid regurgitation, 2 cases of moderate regurgitation, 4 cases of mild-to-moderate regurgitation. Conclusions: For severe tricuspid regurgitation after left-sided valve surgery, the advanced minimally invasive techniques can significantly reduce the operative mortality and morbidity. Minimally invasive bioprosthetic tricuspid valve replacement is a reliable alternative for severe tricuspid regurgitation after left-sided valve surgery.

[Surgical treatment for tricuspid regurgitation after left-sided valve surgery].

Late tricuspid regurgitation after left-sided valve surgery can negatively affect long-term prognosis. The surgical timing and strategy of tricuspid valve reoperation will have important impact on the surgical outcomes. However, there is no clear recommendations of the surgical timing for this condition in the current guidelines. Generally, tricuspid valve reoperation should be performed before irreversible right heart failure occurs. Although tricuspid valve repair is the first choice for tricuspid regurgitation, bioprosthetic tricuspid valve replacement might be a reliable alternative when tricuspid leaflets have severe rheumatic damage or right ventricle and tricuspid annulus significantly dilate. Combined minimally invasive surgical techniques, including right minithoracotomy approach, accessing the right atrium directly through the pericardium with limited dissection, peripheral cannulation strategy with the vacuum-assist single venous drainage technique and heart beating technique, can significantly decrease the operative mortality and postoperative bleeding. With development of interventional therapy, transcatheter tricuspid valve repair or replacement may become alternatives for tricuspid regurgitation after left-sided valve surgery in the future.

Micromachined phase-shifted array-type Mirau interferometer for swept-source OCT imaging: design, microfabrication and experimental validation.

OCT instruments permit fast and non-invasive 3D optical biopsies of biological tissues. However, they are bulky and expensive, making them only affordable at the hospital and thus, not sufficiently used as an early diagnostic tool. Significant reduction of system cost and size is achieved by implementation of MOEMS technologies. We propose an active array of 4x4 Mirau microinterferometers where the reference micro-mirrors are carried by a vertical comb-drive microactuator, enabling the implementation of the phase-shifting technique that improves the sensitivity and eliminates unwanted interferometric terms. We focus on the design of the imaging system, the microfabrication and the assembly of the Mirau microinterferometer, and the swept-source OCT imaging.

[Clinical outcome of allogeneic hematopoietic stem cell transplantation with FLAG sequential busulfan/cyclophosphamide conditioning regimen for refractory/relapsed acute myeloid leukemia].

Objective: To investigate the therapeutic effects of allogeneic hematopoietic stem cell transplantation (allo-HSCT) with FLAG sequential busulfan/cyclophosphamide(Bu/Cy) conditioning regimen for refractory/relapsed acute myeloid leukemia. Methods: From February 2012 to June 2017, 21 patients with refractory/relapsed acute myeloid leukemia underwent allo-HSCT with FLAG sequential Bu/Cy conditioning regimen. Transplantation-related complications and clinical outcome were retrospectively analyzed. Results: After conditioning, no hepatic veno-occlusive disease (VOD) and grade Ⅲ hemorrhagic cystitis occurred. 76.2% (16/21) patients had fever with 4 septicemia. One patient died of septic shock before engraftment. Twenty patients achieved neutrophil engraftment with a median time of 13 days (range, 10 to 21 days). Seventeen patients achieved platelet engraftment with a median time of 18 days (range, 9 to 25 days). The cumulative incidence of acute graft-versus-host disease (aGVHD) was 39.5%, and 3 patients developed grade Ⅲ-Ⅳ aGVHD. Of 19 patients who survived more than 100 days after transplantation, 4 had local chronic graft-versus-host disease (cGVHD). Of 21 patients, the median survival time was 15 months (range, 0.5 to 67 months) post-transplantation. Transplantation-related mortality rate was 28.7%. Leukemia relapse occurred in 4 patients with a median time of 4 months (range, 3 to 8 months) after transplantation. The cumulative relapse rate at 1 year was 21.4%. The 1-year and 3-year overall survival (OS) rates were 60.7% and 54.9% respectively. Log-rank analysis revealed that bone marrow blasts ≥ 20% or extramedullary leukemia before transplantation, poor platelet engraftment and grade Ⅲ-Ⅳ aGVHD were significantly related to shortened OS (P<0.05). Conclusions: Allo-HSCT with FLAG sequential Bu/Cy conditioning regimen in patients with refractory/relapsed myeloid leukemia has acceptable transplantation-related risk and relapse rate. The 1-year and 3-year OS rates are comparable with those in remission patients.

A graphene-like membrane with an ultrahigh water flux for desalination.

An ultrathin nanoporous membrane which combines high water permeability and high salt rejection is the core of ultrafiltration technology. Recently, we reported the synthesis of a chemically robust and nanoporous two-dimensional conjugated aromatic polymer (2D-CAP) membrane. Due to its array of highly regular sub-nanometer pores and channels, the ultrathin 2D-CAP membrane can be potentially used in desalination. Herein, we used molecular dynamics simulations to analyze the transmembrane hydrodynamics of mono- and multi-layer 2D-CAP membranes as a function of layer number. The energy barriers to water and ions across these membranes were calculated to evaluate the potential of 2D-CAP to function as the ultimate RO membrane. Our simulation results show that the bilayer CAP membrane exhibits superior ion rejection (100%) and a water flux (1172 L m-2 h-1 bar-1) with a performance that is three orders of magnitude higher than the commercial reverse osmosis membrane, which is three times higher than the theoretically reported monolayer nanoporous MoS2 membrane (the state-of-the-art membrane reported for desalination). In addition, the 2D-CAP bilayer membrane is highly resistant to swelling even at a high water flux. The monolayer 2D-CAP membrane shows good ion selectivity between monovalent and divalent ions.

Clinical analysis of NSCLC patients reveals lack of association between EGFR mutation and TET1 downregulation.

Lung cancer is one of the leading causes of death from cancer worldwide, with a poor prognosis in advanced cases. In the past decade, epidermal growth factor receptor (EGFR) inhibitors have shown significant efficacy towards treatment for EGFR mutant lung cancer. Expanding our knowledge of oncogenic EGFR signaling pathways is therefore of highly importance for the cancer field. Recently it has been proposed that mutant EGFR transcriptionally silences the TET1 (ten-eleven translocation methylcytosine dioxygenase 1) gene in cellular and animal models of lung cancer. Since TET1 is a known DNA demethylase, EGFR-mediated TET1 silencing therefore downregulates demethylation of tumor suppressor genes, which then leads to tumor growth inhibition, potentiating the role of TET1 as a tumor suppressor gene in NSCLC. In our study, we examined the role of EGFR-TET1 silencing in NSCLC patient samples. By independently analyzing the TCGA (The Cancer Genome Atlas) NSCLC data set as well as a cohort of patient samples from our hospital and a data set from publicly deposited databases, we did not observe the aforementioned mutant EGFR silencing of TET1. Conversely, in our cohort, TET1 expression levels were significantly elevated in EGFR mutant samples (P=0.007). Patients with higher TET1 levels showed a trend of better response rates to EGFR inhibitors compared to low TET1 staining levels, although the result was not significant (P=0.08). Furthermore, we did not observe a correlation between TET1 expression levels and patient survival. We conclude that while oncogenic EGFR suppression of TET1 is established in cellular and animal models of lung cancer, its role in patient outcome and prognosis remains inconclusive and warrants further investigation.

Reduction of progesterone, estradiol and hCG secretion by perfluorooctane sulfonate via induction of apoptosis in human placental syncytiotrophoblasts.

INTRODUCTION: Perfluorooctane sulfonate (PFOS) is widely used as surfactants, lubricants, adhesives, fire retardants and propellants. Animal experiments have shown that PFOS can potentially influence reproductive function. The objective of the present study was to investigate the effects of PFOS on the endocrine function of human placental syncytiotrophoblasts. METHODS: Primary human placental cytotrophoblasts were isolated from term placenta. After syncytialization, the levels of aromatase and apoptosis-related proteins including caspase3, Bcl-2 and Bax were examined after treatment with PFOS from 0.0001 µM to 1 µM or PFOS (0.1 µM) in the presence and absence of apoptosis inhibitor Z-VAD-FMK (30 µM) for 24 h. RESULTS: PFOS suppressed aromatase level and the secretion of estradiol, hCG and progesterone in a concentration-dependent manner from 0.0001 µM to 1 µM with a significant inhibition at 0.001 µM and above in human placental syncytiotrophoblasts. Furthermore PFOS reduced cell viability and induced apoptosis in human placental syncytiotrophoblasts as revealed by increases of pro-apoptosis proteins such as Bax and cleaved-caspase3, and decreases of pro-caspase3 and anti-apoptosis protein Bcl-2. The apoptosis induced by PFOS was further illustrated by increased DNA fragmentation and nuclear condensation. Blocking apoptosis with pan-caspase inhibitor Z-VAD-FMK, the impairment of placental endocrine function by PFOS was restored. DISCUSSION: These results indicate that PFOS may disrupt the secretion of hCG, progesterone and estradiol by human placental syncytiotrophoblasts via induction of apoptosis.

In silico prediction and in vitro identification of bluetongue virus 4 VP5 protein B-cell epitopes.

VP5, the outer capsid protein of bluetongue virus (BTV), plays an important role in viral penetration and antibody-mediated viral neutralization. Therefore, VP5 represents an important target for development of vaccines and diagnostic tests. In this study, we use bioinformatic tools to predict nine antigenic B cell epitopes in the VP5 protein of a BTV serotype 4 (BTV4) isolate from China. Further, we generate five BTV4 VP5-specific monoclonal antibodies (MAbs) and define their corresponding epitopes using a set of VP5-derived peptides expressed as maltose-binding protein (MBP) fusion proteins. The five identified epitopes map to amino acids 119-134, 257-272, 286-301, 322-337, and 481-496 of the VP5 protein. Importantly, the epitopes identified using VP5-derived peptides do not correlate with our bioinformatic prediction of antibody epitopes. Identification and characterization of BTV4 VP5 protein epitopes may aid the development of diagnostic tools and provide information with which to study the structure of the BTV VP5 protein.

Involvement of CRH and hCG in the induction of aromatase by cortisol in human placental syncytiotrophoblasts.

INTRODUCTION: Increased estrogen production in placenta towards the end of gestation plays a pivotal role in the onset of human labor. Estrogen transforms myometrium from a quiescent to a contractile status. Glucocorticoids have been shown to induce estrogen production through the transcription factor specificity protein 1 (Sp1)-mediated induction of aromatase transcription upon elevation of cyclic adenosine mono-phosphate (cAMP) level in human placental syncytiotrophoblasts. However, it is unclear how glucocorticoids activate cAMP pathway thereby inducing aromatase expression in human placental syncytiotrophoblasts. MATERIAL AND METHODS: We investigated this issue in cultured primary human placental syncytiotrophoblasts prepared from placentas collected at term without labor. RESULTS: We demonstrated that cortisol (0.01-1 µM) dose-dependently increased corticotropin-releasing hormone (CRH) and human chorionic gonadotropin (hCG) α/ß subunit expression and their production in the syncytiotrophoblasts. The induction of intracellular cAMP level, Sp1 expression, Sp1 enrichment at the aromatase promoter as well as aromatase expression by cortisol could be partially attenuated by either hCG antibody (1:100) or CRH receptor antagonist α-helical-CRH (1 µM), and further attenuated by combination of hCG antibody and α-helical-CRH. CONCLUSIONS: Cortisol increases aromatase expression via induction of CRH and hCG production and subsequent elevation of cAMP level and enrichment of Sp1 at the aromatase promoter in human placental syncytiotrophoblasts. These findings may account for the parallel increases of cortisol and estrogen production prior to the onset of parturition.

An effective dose of ketamine for eliminating pain during injection of propofol: a dose response study.

BACKGROUND AND PURPOSE: Ketamine can completely eliminate pain associated with propofol injection. However, the effective dose of ketamine to eliminate propofol injection pain has not been determined. The purpose of this study was to determine the effective dose of ketamine needed to eliminate pain in 50% and 95% of patients (ED50 and ED95, respectively) during propofol injections. METHODS: This study was conducted in a double-blinded fashion and included 50 patients scheduled for elective gynecological laparoscopy under general anesthesia. The initial dose of ketamine used in the first patient was 0.25mg/kg. The dosing modifications were in increments or decrements of 0.025 mg/kg. Ketamine was administered 15 seconds before injecting propofol (2.5mg/kg), which was injected at a rate of 1mL/s. Patients were asked to rate their pain during propofol injection every 5s econds using a 0-3 pain scale. The highest pain score was recorded. The ED50, ED95 and 95% confidence intervals (CI) were determined by probit analyses. RESULTS: The dose of ketamine ranged from 0.175 to 0.275 mg/kg. The ED50 and ED95 of ketamine for eliminating pain during propofol injection were 0.227 mg/kg and 0.283 mg/kg, respectively (95%CI: 0.211-0.243 mg/kg and 0.26-0.364 mg/kg, respectively). CONCLUSION: Ketamine at an approximate dose of 0.3mg/kg was effective in eliminating pain during propofol injection.

Identification of a novel bluetongue virus 1-specific B-cell epitope using a monoclonal antibody against the VP2 protein.

Bluetongue virus (BTV) VP2 is an important antigenic protein that can be used for the differential diagnosis of different BTV serotypes. Here, we generated a serotype-specific monoclonal antibody (mab) against BTV1. A series of peptides synthesized based on the amino acid sequence of BTV1 VP2 were screened to define (115)AQPLKVGL(122) as the minimal linear peptide epitope recognized by mab 4B6. Using an immunofluorescence assay (IFA), we found that mab 4B6 reacted strongly with BTV1, but did not react with other BTV serotypes (BTV2-24). The 4B6 will serve as a novel reagent in the development of diagnostic tests for BTV1 infection.