The spleen may be an important target of stem cell therapy for stroke.

Stroke is the most common cerebrovascular disease, the second leading cause of death behind heart disease and is a major cause of long-term disability worldwide. Currently, systemic immunomodulatory therapy based on intravenous cells is attracting attention. The immune response to acute stroke is a major factor in cerebral ischaemia (CI) pathobiology and outcomes. Over the past decade, the significant contribution of the spleen to ischaemic stroke has gained considerable attention in stroke research. The changes in the spleen after stroke are mainly reflected in morphology, immune cells and cytokines, and these changes are closely related to the stroke outcomes. Autonomic nervous system (ANS) activation, release of central nervous system (CNS) antigens and chemokine/chemokine receptor interactions have been documented to be essential for efficient brain-spleen cross-talk after stroke. In various experimental models, human umbilical cord blood cells (hUCBs), haematopoietic stem cells (HSCs), bone marrow stem cells (BMSCs), human amnion epithelial cells (hAECs), neural stem cells (NSCs) and multipotent adult progenitor cells (MAPCs) have been shown to reduce the neurological damage caused by stroke. The different effects of these cell types on the interleukin (IL)-10, interferon (IFN), and cholinergic anti-inflammatory pathways in the spleen after stroke may promote the development of new cell therapy targets and strategies. The spleen will become a potential target of various stem cell therapies for stroke represented by MAPC treatment.

Apatinib exerts anti-tumour effects on ovarian cancer cells.

OBJECTIVE: Apatinib, a small molecule inhibitor of VEGFR-2 tyrosine kinase, shows strong anti-tumour activity against various tumours. The function of apatinib in ovarian cancer, however, remains unclear. This study was conducted to investigate the effects and potential mechanisms by which apatinib modulates the biological function of ovarian cancer cells in vitro and in vivo. METHODS: The effects of apatinib on ovarian cancer cells were determined by assessing cell viability, migration and invasion. The cell cycle distribution and apoptosis of ovarian cancer cells were analysed using flow cytometry. Western blotting was performed to determine the levels of signalling pathway markers. A mouse xenograft model was used to evaluate the efficacy of apatinib in preventing tumour growth. RESULTS: Apatinib did not appreciably affect ovarian cancer cell proliferation and vitality, but did inhibit ovarian cancer cell migration. Apatinib suppressed the epithelial-mesenchymal transition in ovarian cancer cells by inhibiting the JAK/STAT3, PI3K/AKT and Notch signalling pathways. Apatinib effectively inhibited tumour growth in vivo. CONCLUSION: Based on our findings, apatinib is a highly potent, orally active anti-angiogenic and anti-ovarian cancer agent.

Association of chronic hepatitis B virus infection with preterm birth: our experience and meta-analysis.

OBJECTIVES: To assess the association of chronic hepatitis B virus (HBV) infection with preterm birth (PTB). METHODS: A cohort of 20,498 pregnant women (497 HBV carriers with 20,001 non-HBV controls) with normal alanine aminotransferase (ALT) levels was selected from the Obstetrics & Gynecology Hospital of Nantong University. The clinical parameters and PTB incidence were compared between HBV carriers and non-HBV subjects. For the meta-analysis, we searched the PubMed, Ovid and Cochrane Library databases for studies comparing PTB incidence between individuals with chronic HBV infection and non-HBV subjects. RESULTS: HBV carriers were slightly older and had slightly higher ALT levels within normal limits. The body mass index, education and history of pregnancy between HBV carrier and non-HBV groups were comparable. PTB incidence was not associated with HBV carrier status [relative risk (RR) 0.98, 95% confidence interval (CI) 0.71-1.37] in our cohort. However, the meta-analysis involving eight published studies and our study revealed a significant association between chronic HBV infection and PTB incidence (pooled RR 1.26, 95% CI 1.19-1.33). CONCLUSION: While maternal HBV carriers did not have a higher incidence of PTB in our cohort, the meta-analysis indicates that individuals with chronic HBV infection appeared to be at risk of PTB as a whole.

Bicyclol attenuates tetracycline-induced fatty liver associated with inhibition of hepatic ER stress and apoptosis in mice.

Endoplasmic reticulum (ER) stress is known to be involved in the development of several metabolic disorders, including non-alcoholic fatty liver disease (NAFLD). Tetracycline can cause hepatic steatosis, and ER stress may be involved in tetracycline-induced fatty liver. Our previous study showed that bicyclol has been proven to protect against tetracycline-induced fatty liver in mice, and ER stress may also be involved in bicyclol's hepatoprotective effect. Therefore, this study was performed to investigate the underlying mechanisms associated with ER stress and apoptosis, by which bicyclol attenuated tetracycline-induced fatty liver in mice. Bicyclol (300 mg/kg) was given to mice by gavage 3 times. Tetracycline (200 mg/kg, intraperitoneally) was injected at 1 h after the last dose of bicyclol. At 6 h and 24 h after single dose of tetracycline injection, serum ALT, AST, TG, CHO and hepatic histopathological examinations were performed to evaluate liver injuries. Hepatic steatosis was assessed by the accumulation of hepatic TG and CHO. Moreover, hepatic apoptosis and ER stress related markers were determined by TUNEL, real-time PCR, and western blot. As a result, bicyclol significantly protected against tetracycline-induced fatty liver as evidenced by the decrease of elevated serum transaminases and hepatic triglyceride, and the attenuation of histopathological changes in mice. In addition, bicyclol remarkably alleviated hepatic apoptosis and the gene expression of caspase-3, and increased the gene expression of XIAP. The gene expressions of ER stress-related markers, including CHOP, GRP78, IRE-1α, and ATF6, which were downregulated by bicyclol pretreatment in tetracycline-injected mice. These results suggested that bicyclol protected tetracycline-induced fatty liver partly due to its ability of anti-apoptosis associated with ER stress.

Maternal hepatitis B virus carrier status and pregnancy outcomes: a prospective cohort study.

BACKGROUND: Infection with hepatitis B virus (HBV) in pregnant women may be a threat for both mothers and fetuses. This study was performed to explore the impact of maternal HBV carrier status on pregnancy outcomes. METHODS: We conducted a prospective cohort study at the Obstetrics & Gynecology Hospital of Nantong University between January 1, 2012 and September 30, 2015. A consecutive sample of 21,004 pregnant women, 513 asymptomatic HBV carriers and 20,491 non-HBV controls, was included in this study. The main outcomes of interest were selected pregnancy outcomes including miscarriage, stillbirth, preterm birth (PTB), gestational diabetes (GDM), intrahepatic cholestasis of pregnancy (ICP), preterm premature rupture of the membrane (PPROM), low birth weight (LBW), small for gestational age (SGA) and Apgar scores. The incidence of adverse pregnancy outcomes between asymptomatic HBV carriers and non-HBV controls were compared using the chi-square test and logistic regression. P values were two sided, and P <0.05 was considered to indicate statistical significance. RESULTS: The incidences of stillbirth, PTB, GDM, ICP, PPROM, LBW, and SGA were similar between the HBV carrier and non-HBV groups. The proportion of miscarriage was significantly higher among the HBV carriers than the controls (9.36% vs 5.70%; P <0.001). After using multivariate modelling to adjust for possible socio-demographical variables and obstetric complications, women with HBV carrier status were still more likely to have miscarriage (adjusted OR 1.71, 95% CI 1.23-2.38). In addition, the incidences of other maternal and neonatal outcomes were similar between the two groups. CONCLUSION: Maternal HBV carrier status may be an independent risk factor for miscarriage and careful surveillance is warranted.

Transcriptome analysis of food habit transition from carnivory to herbivory in a typical vertebrate herbivore, grass carp Ctenopharyngodon idella.

BACKGROUND: Although feeding behavior and food habit are ecologically and economically important properties, little is known about formation and evolution of herbivory. Grass carp (Ctenopharyngodon idella) is an ecologically appealing model of vertebrate herbivore, widely cultivated in the world as edible fish or as biological control agents for aquatic weeds. Grass carp exhibits food habit transition from carnivory to herbivory during development. However, currently little is known about the genes regulating the unique food habit transition and the formation of herbivory, and how they could achieve higher growth rates on plant materials, which have a relatively poor nutritional quality. RESULTS: We showed that grass carp fed with duckweed (modeling fish after food habit transition) had significantly higher relative length of gut than fish before food habit transition or those fed with chironomid larvae (fish without transition). Using transcriptome sequencing, we identified 10,184 differentially expressed genes between grass carp before and after transition in brain, liver and gut. By eliminating genes potentially involved in development (via comparing fish with or without food habit transition), we identified changes in expression of genes involved in cell proliferation and differentiation, appetite control, circadian rhythm, and digestion and metabolism between fish before and after food habit transition. Up-regulation of GHRb, Egfr, Fgf, Fgfbp1, Insra, Irs2, Jak, STAT, PKC, PI3K expression in fish fed with duckweed, consistent with faster gut growth, could promote the food habit transition. Grass carp after food habit transition had increased appetite signal in brain. Altered expressions of Per, Cry, Clock, Bmal2, Pdp, Dec and Fbxl3 might reset circadian phase of fish after food habit transition. Expression of genes involved in digestion and metabolism were significantly different between fish before and after the transition. CONCLUSIONS: We suggest that the food habit transition from carnivory to herbivory in grass carp might be due to enhanced gut growth, increased appetite, resetting of circadian phase and enhanced digestion and metabolism. We also found extensive alternative splicing and novel transcript accompanying food habit transition. These differences together might account for the food habit transition and the formation of herbivory in grass carp.

Carboxymethylated Rhizoma alismatis polysaccharides reduces the risk of calcium oxalate stone formation by reducing cellular inflammation and oxidative stress.

This study aims to elucidate the mechanism and potential of Rhizoma alismatis polysaccharides (RAPs) in preventing oxidative damage to human renal proximal tubule epithelial cells. The experimental approach involved incubating HK-2 cells with 100 nm calcium oxalate monohydrate for 24 h to establish a cellular injury model. Protection was provided by RAPs with varying carboxyl group contents: 3.57%, 7.79%, 10.84%, and 15.33%. The safeguarding effect of RAPs was evaluated by analyzing relevant cellular biochemical indicators. Findings demonstrate that RAPs exhibit notable antioxidative properties. They effectively diminish the release of reactive oxygen species, lactate dehydrogenase, and malondialdehyde, a lipid oxidation byproduct. Moreover, RAPs enhance superoxide dismutase activity and mitochondrial membrane potential while attenuating the permeability of the mitochondrial permeability transition pore. Additionally, RAPs significantly reduce levels of inflammatory factors, including NLRP3, TNF-α, IL-6, and NO. This reduction corresponds to the inhibition of overproduced pro-inflammatory mediator nitric oxide and the caspase 3 enzyme, leading to a reduction in cellular apoptosis. RAPs also display the ability to suppress the expression of the HK-2 cell surface adhesion molecule CD44. The observed results collectively underscore the substantial anti-inflammatory and anti-apoptotic potential of all four RAPs. Moreover, their capacity to modulate the expression of cell surface adhesion molecules highlights their potential in inhibiting the formation of kidney stones. Notably, RAP3, boasting the highest carboxyl group content, emerges as the most potent agent in this regard.

Carboxymethylated Rhizoma alismatis Polysaccharides Regulate Calcium Oxalate Crystals Growth and Reduce the Regulated Crystals' Cytotoxicity.

OBJECTIVE: This study explored the effects of polysaccharides (RAPD) extracted from the traditional anti-stone Chinese medicine Rhizoma alismatis and their carboxymethylated derivatives (RAPs) on the crystal phase, morphology, and size of calcium oxalate (CaOx). It also determined the damaging ability of the regulated crystals on human renal tubular epithelial cells (HK-2). METHODS: RAPD carboxymethylation with a carboxyl group (-COOH) content of 3.57% was carried out by the chloroacetic acid solvent method. The effects of -COOH content in RAPs and RAP concentration on the regulation of CaOx crystal growth were studied by controlling the variables. Cell experiments were conducted to explore the differences in the cytotoxicity of RAP-regulated crystals. RESULTS: The -COOH contents of RAPD, RAP1, RAP2, and RAP3 were 3.57%, 7.79%, 10.84%, and 15.33%, respectively. RAPs can inhibit the growth of calcium oxalate monohydrate (COM) and induce the formation of calcium oxalate dihydrate (COD). When the -COOH content in RAPs was high, their ability to induce COD formation was enhanced. In the crystals induced by RAPs, a high COD content can lower the damage to cells. In particular, the cytotoxicity of the crystals induced by RAP3 was the lowest. When the concentration of RAP3 increased, the cytotoxicity gradually increased due to the reduced size of the formed COD crystals. An interaction was observed between RAPs and crystals, and the number of RAPs adsorbed in the crystals was positively correlated with the -COOH content in RAPs. CONCLUSIONS: RAPs can reduce the damage of CaOx to HK-2 cells by regulating the crystallization of CaOx crystals and effectively reducing the risk of kidney stone formation. RAPs, especially RAP3 with a high carboxyl group content, has the potential to be developed as a novel green anti-stone drug.

The tissue factor expression on CD14++CD16-monocytes is a new markers in the Chinese Han older adult population with sepsis: A prospective study.

OBJECTIVE: To investigate the tissue factor (TF) expression on different subsets of monocyte and tissue factor secretion of peripheral blood and evaluate their association with the prognosis of sepsis in the Chinese older adult population. METHODS: Monocyte subsets and TF expression on different subsets of monocyte were measured using flow cytometry in 80 older adult sepsis patients and 40 age and sex matched healthy controls. Plasma level of TF was measured using ELISA (enzyme-linked immunosorbent assay) method. RESULTS: TF expression on CD14++CD16- (MO1) monocyte was lower in death (28-day non-survivor) group compared with survival (28-day survivor) groups [1.01 % (0.58 %, 1.62 %) vs. 3.66 % (1.32 %, 6.93 %), p = 0.001]. The plasma level of TF was increased in death group compared to survival group according to the 28-day mortality [109.2 (67.3, 154.2) vs. 62.1 (44.7, 115.5) pg/mL, p = 0.031]. Logistic regression analysis showed TF expression on MO1 monocyte (ß = -0.776, OR = 0.460, CI: 0.251, 0.843, p = 0.012) was independently associated with the 28-day mortality. The ROC (receiver operating characteristic) curve showed that the AUC (area under the curve) of the TF expression on MO1 monocyte for predicting 28-day mortality was 0.846 (p < 0.001). CONCLUSION: The TF expression on CD14++CD16- monocyte is a new marker for the prognosis of older adult sepsis.

Interaction between nanometer calcium oxalate and renal epithelial cells repaired with carboxymethylated polysaccharides.

OBJECTIVE: Injury of renal tubular epithelial cells (HK-2) is an important cause of kidney stone formation. In this article, the repairing effect of polysaccharide (PCP0) extracted from the traditional Chinese medicine Poria cocos and its carboxymethylated derivatives on damaged HK-2 cells was studied, and the differences in adhesion and endocytosis of the cells to nanometer calcium oxalate monohydrate (COM) before and after repair were explored. METHODS: Sodium oxalate (2.8 mmol/L) was used to damage HK-2 cells to establish a damage model, and then Poria cocos polysaccharides (PCPs) with different carboxyl (COOH) contents were used to repair the damaged cells. The changes in the biochemical indicators of the cells before and after the repair and the changes in the ability to adhere to and internalize nano-COM were detected. RESULTS: The natural PCPs (PCP0, COOH content = 2.56%) were carboxymethylated, and three carboxylated modified Poria cocos with 7.48% (PCP1), 12.07% (PCP2), and 17.18% (PCP3) COOH contents were obtained. PCPs could repair the damaged HK-2 cells, and the cell viability was enhanced after repair. The cell morphology was gradually repaired, the proliferation and healing rate were increased. The ROS production was reduced, and the polarity of the mitochondrial membrane potential was restored. The level of intracellular Ca2+ ions decreased, and the autophagy response was weakened. CONCLUSION: The cells repaired by PCPs inhibited the adhesion to nano-COM and simultaneously promoted the endocytosis of nano-COM. The endocytic crystals mainly accumulated in the lysosome. Inhibiting adhesion and increasing endocytosis could reduce the nucleation, growth, and aggregation of cell surface crystals, thereby inhibiting the formation of kidney stones. With the increase of COOH content in PCPs, its ability to repair damaged cells, inhibit crystal adhesion, and promote crystal endocytosis all increased, that is, PCP3 with the highest COOH content showed the best ability to inhibit stone formation.

Clinical significance of neutrophil-lymphocyte ratio and monocyte-lymphocyte ratio in women with hyperglycemia.

Objective: Abnormal pro-inflammatory regulation of the immune system might contribute to the pathogenesis of hyperglycemia during pregnancy. We examined the correlations of neutrophil-lymphocyte ratio (NLR) and monocyte-lymphocyte ratio (MLR) with disease severity and assessed their predictive values. Methods: This retrospective case-control study included 311 cases of hyperglycemia first detected during pregnancy (HFDP) [153 with gestational diabetes mellitus (GDM) and 158 with diabetes in pregnancy (DIP)] and, as a control group, 172 pregnant women with normal glucose tolerance. The NLRs and MLRs were calculated from the blood test data. Results: The absolute leukocyte, neutrophil, monocyte, and lymphocyte counts as well as the NLR and MLR values of HFDP patients significantly differed from control values, but no significant differences were detected in the leukocyte, neutrophil, and monocyte counts of the GDM and DIP groups. Significantly different metrics were selected, binary analysis performed, and odds ratios calculated to identify risk factors. Age, BMI, NLR, and MLR were found to be risk factors for HFDP, and high systolic blood pressure (SBP) at triage and MLR related to the occurrence of DIP. Receiver operating characteristics curve analysis showed that NLR and MLR had better diagnostic accuracy in distinguishing HFDP from controls [NLR area under the curve (AUC) = 0.78; MLR AUC = 0.72] than age and BMI. Values for NLR > 4.394 or MLR > 0.309 correlated with the severity of maternal clinical symptoms and perinatal infant outcomes. MLR was the best predictor of DIP (AUC = 0.72) and MLR values > 0.299 could identify patients at risk for developing DIP and having poor fetal outcomes. Conclusion: Metrics derived from peripheral blood neutrophil, monocyte, and lymphocyte counts are thought to reflect systemic immune-inflammation. Elevated MLR and NLR may be unfavorable prognostic factors for clinical outcomes in patients with hyperglycemia during pregnancy.

Assessment efficacy of neutrophil-lymphocyte ratio and monocyte-lymphocyte ratio in preeclampsia.

OBJECTIVE: Abnormal changes in immune-mediated inflammation contribute to the pathogenesis of preeclampsia (PE). We aim to investigate the value of systemic immune inflammation indices-neutrophil-lymphocyte ratio (NLR) and monocyte-lymphocyte ratio (MLR)-to identify and evaluate the prognosis of patients with PE. METHODS: This study reviewed clinical records of 367 PE patients (162 with mild PE and 205 with severe PE), in addition to a control group of 172 normal pregnancies. Blood cell counts were performed at the first diagnosis of PE, and NLR and MLR were calculated by absolute cell count. RESULTS: Absolute neutrophil, lymphocyte, and monocyte counts and NLR and MLR values in PE were significantly different from controls, although monocyte counts did not significantly differ between mild and severe PE. Receiver operating characteristics curve (ROC) analysis showed NLR and MLR had better diagnostic accuracy in distinguishing PE from controls [NLR area under the curve (AUC) = 0.70; MLR AUC = 0.78]. Further, NLR was the best predictor of disease severity (AUC = 0.71). Cutoff values of NLR > 4.198 or MLR > 0.325 for control and PE groups or a cutoff value of NLR > 4.182 for PE groups indicated that patients were more likely to encounter preterm delivery, have shorter admission-to-delivery interval, and develop maternal and neonatal complications. CONCLUSION: Secondary analyses of white blood cell differential count parameters effectively evaluate the systemic inflammatory/immune state. Compared with absolute cell counts, NLR and MLR offer more effective indicators of clinical assessment, disease severity evaluation, and prognosis evaluation of PE.

[Detecting perfluorooctanoic acid by reversed phase high performance liquid chromatography with pre-column ultraviolet derivatization].

OBJECTIVES: To establish a reversed phase high performance liquid chromatographic method with pre-column ultraviolet derivatization for the determination of perfluorooctanoic acids (PFOA) in sewage and urine. METHODS: PFOA in sewage and urine formed an ion-pair with tetrabutylammonium (TBA) and were extracted into MTBE. The PFOA then reacted with omega-bromoacetophenone (omega-BAP) in acetonitrile and produced materials quantifiable by HPLC with UV detector. RESULTS: The detection limit of the proposed method was 7.0 microg/L with a relative standard deviation of less than 10%. The recoveries for real samples ranged from 85.3% to 116%. CONCLUSION: The proposed method is suitable for the determination of PFOA in sewage and urine.

siRNA­mediated knockdown of T­bet and RORγt contributes to decreased inflammation in pre­eclampsia.

Abnormal immune response resulting from disordered T helper (Th)1/Th2 and Th17/regulatory T cells (Treg) cytokine expression has been demonstrated to serve an important role in the pathogenesis of preeclampsia (PE). However, the role of transcription factors regulating Th cell differentiation contributing to PE remain unclear. To determine whether a decrease in the expression of the T cell lineage transcription factor T­bet can restore immune balance and alleviate the systemic inflammatory response present in PE, 30 patients diagnosed with PE were assessed and compared with healthy pregnant controls. The expression of the transcription factors T­bet and retinoic acid receptor­related orphan receptor (ROR)γt were increased in the peripheral blood mononuclear cells of PE patients compared with controls, consistent with the presence of abnormally high T­bet:GATA3 and RORγt:forkhead box (FOX) P3 ratios. The present study additionally identified a high­efficiency, specific small interfering (si)RNA that can downregulate RORγt and T­bet mRNA levels and inhibit protein expression. This effective siRNA was transfected into activated CD4+ T cells derived from patients with PE to observe the changes to transcription factor expression and attempt to elucidate the regulatory mechanism of T cell subsets. It was identified that knockdown of RORγt induced increased expression of FOXP3 and that the ratios of RORγt:FOXP3 and interleukin (IL)­17A:IL­10 were subsequently decreased. The results suggested that siRNA­mediated knockdown of T­bet regulated the immune balance of Th17/Tregs via changes to RORγt and FOXP3. When siRNA against RORγt and T­bet were used in combination, a stronger ability to regulate immune balance was observed. These results imply that Th1­ and Th17­type immunity is dominant in PE and that the siRNA­mediated knockdown of certain Th1 and Th17 cell transcription factors may be an effective therapeutic target for promoting immune balance in CD4+ T cell subgroups and ameliorating local and generalized inflammation in PE.

Liver Dysfunction during Pregnancy and Its Association of With Preterm Birth in China: A Prospective Cohort Study.

BACKGROUND: Liver dysfunction is common in pregnancy but its association with adverse pregnancy outcomes such as preterm birth (PTB) remains unclear. METHODS: A prospective cohort of HBV-infected or uninfected pregnant women attending antenatal care was recruited at Nantong Maternal and Child Health Hospital between January 1, 2012, and June 30, 2016. Liver function tests (LFTs) were monitored through pregnancy. The primary outcomes were PTB and very PTB (delivery prior 37 and 32weeks' gestation respectively). Poisson regression was used to estimate adjusted risk ratios (RR) for women with HBV infection and LFT abnormalities. RESULTS: Among 36,755 pregnant women (1,113 HBV carriers and 35,642 non-HBV subjects), 3,519 (9.57%) had abnormal LFTs. The commonest cause for liver dysfunction during pregnancy was non-alcoholic fatty liver diseases (NAFLD, 51.3%). Abnormal aspartate aminotransferase (AST), gamma-glutamyltransferase (GGT) and two folds upper limit of normal total bilirubin (RR and 95%CI: 2.73, 1.30-5.76; 2.24, 1.35-3.31; 2.01, 1.22-3.31 respectively), rather than HBsAg positivity, were identified as independent risk factors for preterm birth. Besides, GGT abnormality was associated with increased risk of very PTB. CONCLUSIONS: We suggest that surveillance of LFTs among pregnant women should be warranted, given the increased risk of PTB.

Butorphanol suppresses fentanyl-induced cough during general anesthesia induction: A randomized, double-blinded, placebo-controlled clinical trial.

Fentanyl-induced cough (FIC) is unwanted in the patients requiring stable induction of general anesthesia. This study was designed to evaluate the suppressive effects of butorphanol pretreatment on the incidence and severity of FIC during the induction of general anesthesia. A total of 315 patients of American Society of Anesthesiologists physical status I and II, scheduled for elective surgery under general anesthesia were randomized into 3 equally sized groups (n = 0105). Two minutes before fentanyl bolus, group I received intravenously 5 mL normal saline, groups II and III received butorphanol 0.015 and 0.03 mg/kg (diluted with saline to 5 mL), respectively. Patients were then administrated with fentanyl 2.5 µg/kg within 5 s. The incidence and severity of FIC was recorded for 2 minutes after fentanyl bolus. During experimental period, the mean arterial pressure, heart rate, and peripheral capillary oxygen saturation (SpO2) were recorded before the administration of butorphanol or normal saline (T0), 2 minutes (T1) after butorphanol injection, and 2 minutes (T2) after fentanyl injection. The incidence of FIC was 31.4% in group I, 11.4% in group II, and 3.8% in group III. Group III had a lowest incidence of FIC among 3 groups (P < 0.001, vs group I; P < 0.05, vs group II). The severe FIC was not observed in groups II and III, but was recoded from 6 patients in group I. At 2 minutes after fentanyl injection (T2), the mean arterial pressure was significantly higher in group I than that in groups II and III (P < 0.01, vs group II; P < 0.05, vs group III), but the values remained within safe limits. In conclusion, pretreatment with butorphanol could effectively and safely suppress FIC during anesthesia induction.

[Establishment of the human acute promyelocytic leukemia pathological model in the SCID beige mice by using NB4 cell line].

OBJECTIVE: This study was to establish a stable, effective and reproducible human acute promyelocytic leukemia model in severe combined immunodeficient (SCID) mice by using NB4 cell line, and to investigate the disease course character and biological behaviors. METHODS: Three-five-week-old SCID beige mice were divided randomly into two groups: experimental and control group. SCID mice of experimental group were transplanted by tail vein (iv) injection of 5×10(6) NB4 cells. The WBC cell count and the positive rate of promyelocytes in peripheral blood were dynamically monitored by using smears. Morphological examination and histopathological assay were employed to confirm NB4 cell infiltration in organs (liver, spleen, lung, kidney and brain). The expression level of PML-RARα fusion protein was detected by Western blot. RESULTS: Within two weeks there was no significant difference in peripheral blood WBC count between two groups (P > 0.05), meanwhile, NB4 cells were not found. At the day 21 and 28 after inoculation, the peripheral blood white blood cell count of experimental group reached to (4.79 ± 1.13)×10(9)/L and (7.62 ± 2.24)×10(9)/L respectively, which were significantly higher than that in control group (P < 0.05); simultaneously, the positive rates of promyelocytes on smears were (2.14 ± 0.63)% and (6.6 ± 2.76)%, respectively. Morphological observation showed single or multiple tumor lumps at day 21 after inoculation; HE staining of tissue biopsies demonstrated a large number of promyelocyte in the liver, spleen, lung, kidney and brain tissue. Cell immunofluorescence results showed that the CD33 expression of bone marrow cells in mice of experimental group was strongly positive (P < 0.05). Western blot confirmed that the PML-RARα fusion protein was expressed variously in liver, kidney and brain tissue. CONCLUSIONS: The human acute promyelocytic leukemia SCID mouse model is succesfully established by tail vein injection of NB4 cells. This model can mimic the characters of involved bone marrow and diffuse growth of cells. This model is a useful tool to explore the pathogenic mechanism and experimental treatment of human leukemia.

Molecular cloning, expression and single nucleotide polymorphisms of protein phosphatase 1 (PP1) in mandarin fish (Siniperca chuatsi).

In the wild, mandarin fish (Siniperca chuatsi) only feed on live prey fish, refusing dead prey. When reared in ponds, training will result in some mandarin fish accepting artificial diets. However, little is currently known about the molecular mechanism of the individual difference. Serine/threonine protein phosphatase 1 (PP1) is a suppressor of learning and long-term memory (LTM) in mammals. In the present study, the relationship between PP1 and the individual difference in acceptance of artificial diets in mandarin fish was investigated. The complete CDS (coding sequence) of four PP1 isoforms (PP1caa, PP1cab, PP1cb and PP1cc) were cloned in mandarin fish. The amino acid sequences of these PP1 isoforms are highly conserved in different species. The mRNA expressions of PP1caa and PP1cb in brain of artificial diet feeders were significantly higher than those in nonfeeders, suggesting the deficiency in the maintenance of long-term memory of its natural food habit (live prey fish). The SNP loci in PP1caa and PP1cb were also found to be associated with the individual difference in acceptance of artificial diets in mandarin fish. These SNPs of PP1caa and PP1cb genes could be useful markers for gene-associated breeding of mandarin fish, which could accept artificial diets. In conclusion, different mRNA expression and SNPs of PP1caa and PP1cb genes in feeders and nonfeeders of artificial diets might contribute to understanding the molecular mechanism of individual difference in acceptance of artificial diets in mandarin fish.

Inhibitory effects of the recombinant human serine protease inhibitor Hespintor on the proliferation, migration and invasion of hepatocellular carcinoma cells.

Hespintor is a new Kazal-type serine proteinase inhibitor (Serpin) screened from the HepG2 hepatoblastoma cell line using the suppression subtractive hybridization (SSH) technique. Seprin is closely associated with the progression and remission of malignant tumors, and has certain significance in the diagnosis and treatment of tumors. Investigations on the antitumor activity of Serpin are expected to aid in the development of a new method for tumor treatment based on the serine protease inhibitor. Although the Hespintor prokaryotic expression strain and recombinant Hespintor protein (recombinant fusion protein of Hespintor and rHespintor) have already been obtained, the protein extraction efficiency is low due to the low initial amount of extracted protein and large number of purification steps, which affect the study of the protein function. The aim of the present study was to improve the purification method of rHespintor, increase the protein extraction efficiency, and investigate its effects on the proliferation, migration and invasion of the HepG2 hepatoblastoma cell line. The results demonstrated that the application of urea gradient washing of inclusion body of the protein may effectively remove the majority of impure proteins from the targeted protein. After one-step purification, the target protein rHespintor exhibited a high inhibitory effect of Trypsin Hydrolysis, which was exhibited in a dose-dependent manner. Hoechst 33258 staining was used to determine cell apoptosis. After treating HepG2 hepatoblastoma cells with rHespintor, the cell growth was inhibited, the proliferation ability was reduced, and the number of migrated and invaded cells were significantly decreased. Hoechst 33258 staining and flow cytometry assay results showed clear cell apoptosis. The results reveal showed that rHespintor significantly inhibited proliferation, migration and invasion of the HepG2 hepatoblastoma cell line in vitro, and induced cell apoptosis to a certain extent, indicating that the recombinant protein Hespintor exerts an antitumor effect in vitro, and has the potential and feasibility to become an antitumor drug.

Vascular endothelial growth factor affects dendritic cell activity in hypertensive disorders of pregnancy.

Vascular endothelial growth factor (VEGF) activity is involved in the growth and stability of the placenta, and its signaling has been implicated in the development of pregnancy­induced hypertension (PIH). The present study sought to evaluate VEGF levels and dendritic cell (DC) profiles in patients with PIH, and to investigate the effects of VEGF expression on DC phenotypes. The present study assessed 162 patients, 112 of whom were diagnosed with PIH. Serum VEGF was measured by ELISA, while myeloid DC (mDC; (Lin1­HLA­DR+CD11c+) and plasmacytoid DC (pDC; Lin1­HLA­DR+CD123+) counts were determined using flow cytometry. In order to determine the effect of VEGF treatment on DC phenotypes, immature DCs (iDCs) were separated from monocytes by culturing in the presence of cytokines (GM­CSF, IL­4), and then pretreated with VEGF or lipopolysaccharide (LPS). The phenotype of dendritic cells (CD14, CD80, CD83, or CD86) was determined by flow cytometry. The levels of VEGF in the serum of patients with PIH were significantly lower than those in control subjects (P<0.05). The percentage of pDCs found in the serum of patients with preeclampsia was significantly lower than that in the other groups. The percentage of mDCs in the serum of patients with preeclampsia and eclampsia was significantly higher than that in the control and hypertensive disorder groups (P<0.05). The percentage of mDCs was significantly negatively correlated with the levels of VEGF in the preeclamptic and eclamptic patients (r=­0.34 and r=­0.42, respectively; P<0.05). Detected levels of CD80, CD83 and CD86 in DCs treated with VEGF were significant lower than those in DCs treated with LPS alone (P<0.05). In conclusion, abnormal expression of VEGF and an altered dendritic cell profile may be involved in the development of PIH.