Incidence and correlates of insomnia and its impact on health-related quality of life among Chinese pregnant women: a cross-sectional study.

OBJECTIVE: The aims of this study were to examine the incidence and correlates of insomnia and its impact on health-related quality of life among Chinese pregnant women. METHOD: A cross-sectional study was performed from November 2018 to April 2019 in a university-affiliated general hospital in Guangzhou, China. Seven hundred and seventeen pregnant women completed the 36-item Short-Form Health Survey (SF-36), the Self-Rating Anxiety Scale (SAS), the Self-Rating Depression Scale (SDS), and the obstetric and sociodemographic data sheet. FINDINGS: 24.3% of the pregnant women suffered from insomnia. Compared with women without insomnia, those with insomnia had a significantly lower health-related quality of life during pregnancy. Maternal age, educational level, occupation, economic status, insurance coverage, gestational age, the woman's relationship with her mother-in-law and anxiety were significantly associated with insomnia among pregnant women. CONCLUSION: The incidence of insomnia among pregnant women is high, and insomnia is negatively correlated with health-related quality of life. Appropriate measures and practical therapeutic programmes should be provided to prevent the adverse effects of insomnia in pregnant women with advanced maternal age, lower education, lower economic status, unemployment, lack of insurance coverage, unsatisfied with their relationships with their mothers-in-law, and suffering from anxiety symptoms, especially in the third trimester.

Anxiety symptoms and health-related quality of life in mainland Chinese pregnant women: a cross-sectional study.

OBJECTIVE: Health-related quality of life allows the health care professionals to envisage new axes of improvement in antenatal care and is a core aspect of contemporary maternity care provision. This study aims to estimate the prevalence of anxiety symptoms and explore the relationship between anxiety symptoms and health-related quality of life among Chinese pregnant women. METHODS: A cross-sectional study was conducted in a local teaching hospital in Guangzhou, China between April and June, 2018. Seven hundred and seventy Chinese pregnant women completed the 36-Item Short-Form Health Survey (SF-36), the Self-rating Anxiety Scale (SAS) and socio-demographic questionnaires. RESULTS: 18.2% women were classified as having elevated anxiety symptoms as evidenced by a SAS score ≥50. Compared with women without anxiety symptoms, the pregnant women with anxiety symptoms had worse physical (SF36-PCS) and mental (SF36-MCS) health-related quality of life and a lower level of seven domains of SF-36 (GH, RP, BP, VT, SF, RE and MH). Elevated anxiety symptoms predicted worse physical (SF36-PCS) and mental (SF36-MCS) health-related quality of life. The third trimester predicted a lower level of physical (SF36-PCS) health-related quality of life, while an unsatisfied relationship with mother-in-law predicted a lower level of mental (SF36-MCS) health-related quality of life. CONCLUSIONS: The pregnant women with anxiety symptoms had impaired health-related quality of life. Health care professionals should identify pregnant women with anxiety symptoms and facilitate their treatment, which could improve their health-related quality of life.

Health related quality of life in Chinese pregnant women at advanced maternal age: a cross-sectional study.

OBJECTIVE: This study aims to explore the relationship between advanced maternal age (AMA) and health-related quality of life in Chinese pregnant women. METHODS: A cross-sectional study was conducted in Guangzhou, China between September 2018 and June 2019. Four hundred and twenty-seven AMA women and the equal number of their younger counterparts completed the 36-Item Short-Form Health Survey (SF-36). RESULTS: Compared with younger women, the AMA women were more likely to be employed; have a higher monthly household income and insurance covered; have a satisfied relationship with their husband and mother-in-law; and had a significantly lower level of physical (SF36-PCS) health-related quality of life and a higher level of mental (SF36-MCS) health-related quality of life during the pregnancy. The association of maternal age with health-related quality of life varies according with the trimester of pregnancy. Maternal age was a significant predictor of SF36-PCS and SF36-MCS. The third trimester was the significant predictor of SF36-PCS while the relationship with the mother-in-law was the significant predictor of SF36-MCS. CONCLUSIONS: The SF36-PCS in the AMA women decreased with advancing age. However, their SF36-MCS was better over their younger counterparts. Age-related biological disadvantages may be offset by social/psychological advantages in AMA women.

Lysophosphatidic acid enhances human umbilical cord mesenchymal stem cell viability without differentiation via LPA receptor mediating manner.

Human umbilical cord mesenchymal stem cells (hUC-MSCs) are potential stromal cells which are regarded as the most feasible stem cell group in cell therapy. The maintenance of cell survival without differentiation is important in cell transplantation and stem cell therapy. However, negative factors exist in cell transplantation. Lysophosphatidic acid (LPA) is a non-antigenic small molecule phospholipid which induced several fundamental cellular responses, such as cell proliferation, apoptosis and migration. In this study we aimed to explore the effects of LPA on the survival and differentiation of MSCs and its availability in cell therapy. We found that LPA stimulated hUC-MSC proliferation and protected hUC-MSCs from lipopolysaccharide (LPS) induced apoptosis. We also observed that CD29, CD44, CD73, CD90 and CD105 were expressed, whereas CD34 and CD45 were not expressed in hUC-MSCs, and these makers have no change in LPA containing medium, which indicated that LPA accelerated the survival of hUC-MSCs in an undifferentiating status. We also demonstrated that higher expressed LPAR1 involved in LPA stimulated cell survival action. LPA stimulated cell proliferation was associated with LPAR1 mediated Gi/o-proteins/ERK1/2 pathway. On the other hand, LPA protected hUC-MSCs from LPS-induced apoptosis through suppressing caspase-3 activation by LPAR1 coupled with a G protein, but not Gi/o or Gq/11 in hUC-MSC. Collectively, this study demonstrated that LPA increased the proliferation and survival of hUC-MSCs without differentiation through LPAR1 mediated manner. Our findings provide that LPA as a anti-apoptotic agent having potential application prospect in cell transplantation and stem cell therapy.

Astragalus polysaccharide stimulates glucose uptake in L6 myotubes through AMPK activation and AS160/TBC1D4 phosphorylation.

AIM: To establish the mechanism responsible for the stimulation of glucose uptake by Astragalus polysaccharide (APS), extracted from Astragalus membranaceus Bunge, in L6 myotubes in vitro. METHODS: APS-stimulated glucose uptake in L6 myotubes was measured using the 2-deoxy-[(3)H]-D-glucose method. The adenine nucleotide contents in the cells were measured by HPLC. The phosphorylation of AMP-activated protein kinase (AMPK) and Akt substrate of 160 kDa (AS160) was examined using Western blot analysis. The cells transfected with 4P mutant AS160 (AS160-4P) were constructed using gene transfer approach. RESULTS: Treatment of L6 myotubes with APS (100-1600 µg/mL) significantly increased glucose uptake in time- and concentration-dependent manners. The maximal glucose uptake was reached in the cells treated with APS (400 µg/mL) for 36 h. The APS-stimulated glucose uptake was significantly attenuated by pretreatment with Compound C, a selective AMPK inhibitor or in the cells overexpressing AS160-4P. Treatment of L6 myotubes with APS strongly promoted the activation of AMPK. We further demonstrated that either Ca(2+)/calmodulin-dependent protein kinase kinase ß (CaMKKß) or liver kinase B1 (LKB1) mediated APS-induced activation of AMPK in L6 myotubes, and the increased cellular AMP: ATP ratio was also involved. Treatment of L6 myotubes with APS robustly enhanced the phosphorylation of AS160, which was significantly attenuated by pretreatment with Compound C. CONCLUSION: Our results demonstrate that APS stimulates glucose uptake in L6 myotubes through the AMP-AMPK-AS160 pathway, which may contribute to its hypoglycemic effect.

HMGB1 Upregulates RAGE to Trigger the Expression of Inflammatory Factors in the Lung Tissue in a Hypoxic Pulmonary Hypertension Rat Model.

Hypoxic pulmonary hypertension (HPH), a form of pulmonary hypertension (PH) caused by hypoxia, could cause serious complications and has a high mortality rate, and no clinically effective treatments are currently available. In this study, we established an HPH preclinical model in rats by simulating clinicopathological indicators of the disease. Our results showed that high mobility group box-1 protein (HMGB1) aggravated the clinical symptoms of HPH. We aimed at establishing protocols and ideas for the clinical treatment of HPH by identifying downstream HMGB1 binding receptors. Our investigation showed that continuous hypoxia could cause significant lung injury in rats. ELISA and western blotting experiments revealed that HPH induces inflammation in the lung tissue and increases the expression of a hypoxia-inducible factor. Testing of lung tissue proteins in vivo and in human pulmonary artery endothelial cells in vitro revealed that the HMGB1-mediated increase in the receptor for advanced glycation end products (RAGE) expression promoted inflammation. In summary, we successfully established an HPH rat model providing a new model for preclinical HPH research and elucidated the role of HMGB1 in HPH. Furthermore, we describe that HMGB1 induced inflammation in the HPH model via RAGE, causing severe lung dysfunction. This study could potentially provide novel ideas and methods for the clinical treatment of HPH.

Clinical implications for girdin protein expression in breast cancer.

Girdin is highly expressed in breast carcinomas. Suppression of Girdin inhibited breast cancer cell migration. However, the clinical implications of Girdin as a marker are still unclear. Here we examined 80 breast cancer specimens using immunohistochemistry. Overall, positive Girdin staining was 41.25% in all of the cases. Girdin was strongly expressed in tumors of CerbB2-positive breast cancers (p < .05). Cases with both CerbB2- and Girdin-positive expression had a higher histological grade than the others. These findings indicated the closed relationship between breast cancer progression and Girdin expression. Girdin together with CerbB2 might be a new potential marker for breast cancers.

Multiple organ invasion by viruses: pathological characteristics in three fatal cases of the 2009 pandemic influenza A/H1N1.

To further understand the pathological characteristics of multiple organ involvement of the 2009 pandemic influenza A/H1N1 infection, tissues of bronchial mucosa, lung, myocardium, gastrocnemius, and liver from 3 patients with fatal A/H1N1 infections were investigated by light microscopy and transmission electron microscopy. In all 3 patients, bronchial mucosa showed necrotizing bronchiolitis, epithelial necrosis and desquamation, and squamous metaplasia, while lung consolidation or fibrosis was identified. Myocardium and gastrocnemius exhibited focal necrosis and fibrosis, surrounded by muscle cells showing features of cell damage. In liver, there was widespread fatty degeneration and necrosis, most often around the central lobular vein and portal area. Viral particles were found in all samples, frequently located in endothelium, epithelium, and muscle cells. The observations demonstrate that in fatal cases of A/H1N1 infection, viruses not only infect the respiratory system, but also engage in multiple organ invasions, causing pathologic changes.

Role of PKCzeta translocation in the development of type 2 diabetes in rats following continuous glucose infusion.

AIM: We investigated the molecular mechanisms of hyperglycaemia-induced insulin resistance and type 2 diabetes in rats receiving a continuous glucose infusion (GI). METHODS: Female Wistar rats were infused with either 2.8 mol/L glucose or saline (2 mL/h) for durations varying from 0 to 15 days. Blood samples were analysed daily to determine glucose and insulin dynamics. Subsets of animals were sacrificed and soleus muscles were extracted for determination of protein expression, subcellular location, and activities of insulin-signalling proteins. RESULTS: Rats accommodated this systemic glucose oversupply and developed insulin resistance on day 5 (normoglycaemia/hyperinsulinaemia) and type 2 diabetes on day 15 (hyperglycaemia/normoinsulinaemia). The effect of GI on protein kinase Czeta (PKCzeta) activity was independent of changes in phosphatidylinositol 3-kinase activity, and occurred in parallel with an increase in PDK1 activity. Activated PKCzeta was mainly located in the cytosol after 5 days of GI that was coincident with the translocation of GLUT4 to the plasma membrane, and normoglycaemia. After 15 days of GI, PKCzeta translocated from the cytosol to the plasma membrane with a concomitant decrease in PDK1 activity. This caused an increase in the association between PKCzeta and PKB and a decrease in PDK1-PKB reactions at the plasma membrane, leading to reduced PKB activity. The activity of PKCzeta per se was also compromised. The PKCzeta and PKB activity reduction and the blunted insulin-stimulated GLUT4 translocation eventually led to hyperglycaemia and diabetes. CONCLUSION: Translocation of PKCzeta may play a central role in the development of type 2 diabetes.

EXEL-7647 inhibits mutant forms of ErbB2 associated with lapatinib resistance and neoplastic transformation.

PURPOSE: Mutations associated with resistance to kinase inhibition are an important mechanism of intrinsic or acquired loss of clinical efficacy for kinase-targeted therapeutics. We report the prospective discovery of ErbB2 mutations that confer resistance to the small-molecule inhibitor lapatinib. EXPERIMENTAL DESIGN: We did in vitro screening using a randomly mutagenized ErbB2 expression library in Ba/F3 cells, which were dependent on ErbB2 activity for survival and growth. RESULTS: Lapatinib resistance screens identified mutations at 16 different ErbB2 amino acid residues, with 12 mutated amino acids mapping to the kinase domain. Mutations conferring the greatest lapatinib resistance cluster in the NH2-terminal kinase lobe and hinge region. Structural computer modeling studies suggest that lapatinib resistance is caused by multiple mechanisms; including direct steric interference and restriction of conformational flexibility (the inactive state required for lapatinib binding is energetically unfavorable). ErbB2 T798I imparts the strongest lapatinib resistance effect and is analogous to the epidermal growth factor receptor T790M, ABL T315I, and cKIT T670I gatekeeper mutations that are associated with clinical drug resistance. ErbB2 mutants associated with lapatinib resistance transformed NIH-3T3 cells, including L755S and T733I mutations known to occur in human breast and gastric carcinomas, supporting a direct mechanism for lapatinib resistance in ErbB2-driven human cancers. The epidermal growth factor receptor/ErbB2/vascular endothelial growth factor receptor inhibitor EXEL-7647 was found to inhibit almost all lapatinib resistance-associated mutations. Furthermore, no ErbB2 mutations were found to be associated with EXEL-7647 resistance and lapatinib sensitivity. CONCLUSIONS: Taken together, these data suggest potential target-based mechanisms of resistance to lapatinib and suggest that EXEL-7647 may be able to circumvent these effects.

Astragalus polysaccharides alleviates glucose toxicity and restores glucose homeostasis in diabetic states via activation of AMPK.

AIM: To establish the mechanism underlying the improvement of glucose toxicity by Astragalus polysaccharide (APS), which occurred via an AMP activated protein kinase (AMPK)-dependent pathway. METHODS: In vivo and in vitro effects of APS on glucose homeostasis were examined in a type 2 diabetes mellitus (T2DM) rat model. The T2DM rat model was duplicated by a high-fat diet (58% fat, 25.6% carbohydrate, and 16.4% protein) and a small dose of streptozotocin (STZ, 25 mg/kg, ip). After APS therapy (700 mg.kg(-1).d(-1), ig) for 8 weeks, blood glucose, glycosylated hemoglobin, and serum insulin were measured. Insulin sensitivity was evaluated by the comprehensive analysis of oral glucose tolerance tests (OGTT) and HOMA IR index. Hepatic glycogen was observed by the PAS staining method. The expression and activity of skeletal muscle AMPKalpha and acetyl-CoA carboxylase (ACC), and the phosphorylation of hepatic glycogen synthase (GS), the glycogen synthase (GS),were measured by Western blotting. Glucose uptake was measured with the 2-deoxy-[(3)H]-D-glucose method in C2C12 cells. RESULTS: The hyperglycemia status, insulin sensitivity, glucose uptake, and activation level of AMPK in diabetic rats were improved in response to APS administration. APS could also alleviate glucose toxicity in cultured mouse cells by the activation of AMPK. CONCLUSION: APS can alleviate glucose toxicity by increasing liver glycogen synthesis and skeletal muscle glucose translocation in the T2DM rat model, via activation of AMPK.

[Effects of hypoxia on the expression of HMGB1 and related inflammatory factors in human pulmonary artery smooth muscle and pulmonary artery endothelial cells].

OBJECTIVE: To investigate the effects of hypoxia on the expressions of high mobility group box-1(HMGB1), HMGB1 receptors and inflammatory factors in human pulmonary artery smooth muscle cell( HPASMC) and human pulmonary artery endothelial cells (HPAEC).The effects of HMGB1 on the proliferation and migration activity of the two kinds of cells were detected. METHODS: HPASMC and HPAEC were cultured under hypoxic conditions (Hypoxia at 1% oxygen, Hypoxia group) and normoxic conditions(Control group) . The mRNA levels of HMGB1, Toll-like receptors 2,4,9 (TLR2 , TLR4, TLR9), the receptor of advanced glycation end products(RAGE) , CD24 and IL-6 ,TNF-α,CXCL8 were detected by real-time PCR. Cell proliferation was measured by MTS. Cell migration was analysed by wound healing test. RESULTS: Compared with control group, the expressions of HMGB1 mRNA and RAGE mRNA in both HPASMC and HPAEC were increased significantly (P＜0.05 and 0.01). Meanwhile, the expressions of CD24 mRNA in HPAEC and IL-6 mRNA in HPASMC of hypoxia group were increased significantly (P＜0.05). MTS results showed that HMGB1 inhibited the proliferation of HPAEC at 345 pmol/L significantly (P＜0.01). HMGB1 had no effect on the proliferation of HPASMC. Wound healing test showed that HMGB1 had no significant effect on HPASMC and HPAEC. CONCLUSION: HMGB1 was produced by HPAEC and HPASMC induced by hypoxia. HMGB1 induces endothelial barrier dysfunction by inhibiting HPAEC proliferation. Hypoxia stimulates HPASMC to produce inflammatory cytokines.

[Effect of reduced nitrogen fertilization on carbon footprint in spring maize-late rice production system]. / 减氮施肥对春玉米-晚稻生产系统碳足迹的影响.

With the growing concerns on global climate change and food security, low carbon agriculture in food production attracts more attention. Low carbon agriculture needs to balance higher-level crop yields and lower greenhouse gas emission in production process. Improving nitrogen mana-gement may help mitigate greenhouse gas emission and achieve stable or higher crop yields in crop production systems. In this study, we investigated the effects of nitrogen application rates (150, 225, 300 kg N·hm-2) on the carbon footprint of spring maize-late rice rotation system in paddy field using the life cycle assessment. The results showed that greenhouse gas emission and carbon footprint increased with the nitrogen fertilizer application rates in both crops. Nitrogen fertilizer was the most important contributor to carbon footprint of spring maize ecosystem, accounting for 36.2%-50.2%. Methane emission increased with nitrogen fertilizer input and contributed the most to the carbon footprint of late rice production, accounting for 42.8%-48.0%. When the nitrogen application rate was reduced by 25% (225 kg N·hm-2) and 50% (150 kg N·hm-2), greenhouse gas emission of maize production decreased by 21.9% and 44.3%, and the carbon footprint decreased by 20.3% and 39.1%, respectively. Meanwhile, the greenhouse gas emissions of late rice decreased by 12.3% and 20.4%, and the carbon footprint of late rice decreased by 13.7% and 16.7%, respectively. The reduction of nitrogen fertilizer rate had no significant effect on maize yield, with the treatment of 225 kg N·hm-2 rate holding the highest yield in late rice ecosystem. The treatment of 150 kg N·hm-2 rate in spring maize production and 225 kg N·hm-2 rate in late rice production was the sustainable N fertilizer application rate for achieving high grain yield and reducing the carbon footprint in crop system.

Efficient generation of CD34+ progenitor-derived dendritic cells from G-CSF-mobilized peripheral mononuclear cells does not require hematopoietic stem cell enrichment.

As a result of their potent antigen-presentation function, dendritic cells (DC) are important tools for cell therapy programs. In vitro-generated DC from enriched CD34+ hematopoietic stem cells (HSC; enriched CD34 DC) have already proven their efficiency in Phase I/II clinical trials. Here, we investigated whether enrichment of CD34+ HSC before the onset of culture was absolutely required for their differentiation into DC. With this aim, we developed a new two-step culture method. PBMC harvested from G-CSF-mobilized, healthy patients were expanded for 7 days during the first step, with early acting cytokines, such as stem cell factor, fetal liver tyrosine kinase 3 ligand (Flt-3L), and thrombopoietin. During the second step, expanded cells were then induced to differentiate into mature DC in the presence of GM-CSF, Flt-3L, and TNF-alpha for 8 days, followed by LPS exposure for 2 additional days. Our results showed that the rate of CD34+/CD38+/lineageneg cells increased 19.5+/-10-fold (mean+/-sd) during the first step, and the expression of CD14, CD1a, CD86, CD80, and CD83 molecules was up-regulated markedly following the second step. When compared with DC generated from enriched CD34+ cells, which were expanded for 7 days before differentiation, DC derived from nonenriched peripheral blood stem cells showed a similar phenotye but higher yields of production. Accordingly, the allogeneic stimulatory capacity of the two-step-cultured DC was as at least as efficient as that of enriched CD34 DC. In conclusion, we report herein a new two-step culture method that leads to high yields of mature DC without any need of CD34+ HSC enrichment.

[A clinicopathologic study on complications of orthotopic liver transplantation in 54 patients with chronic hepatitis B virus infection].

OBJECTIVE: To study the complication incidence of 54 patients with chronic HBV infection following their orthotopic liver transplantation (OLT), and factors associated with HBV recurrence and hepatocellular carcinoma (HCC) recurrence or metastasis post-OLT. METHODS: The light-microscopic appearance of hepatic allograft biopsies in 54 patients with chronic HBV infection following OLT was examined. The related clinical data were analyzed. The incidence and occurrence time of post-OLT complications were studied. Furthermore, the relationship between recurrent type B viral hepatitis and acute rejection and the relationship among HCC recurrence/metastasis, acute rejection, the tumor diameter, and the portal vein invasion were particularly studied. RESULTS: The frequent complications of patients with chronic HBV infection following OLT were acute rejection [38(70.4%); occurrence time: 5-365 d], chronic rejection [ 1(1.9%); occurrence time: 10.7 months],bile duct complications [24(44.4%); occurrence time: 7-940 d], HBV recurrence [7(13.0%); occurrence time: 1-540 d], HCV infection [3(5.6%); occurrence time: 60 d, 60 d, 33 months], CMV infection [8(14.8%); occurrence time: 67-90 d], and HCC recurrence or metastasis [17(31.5%); occurrence time: 2-41 months]. At the end of 1 year post-OLT, 95% of patients with post-hepatitis B cirrhosis were alive; At the end of 3 years post-OLT, 85% of patients with post-hepatitis B cirrhosis were alive. However, at the end of 1 year post-OLT, 67.6% of patients with post-hepatitis B HCC were alive; At the end of 3 years post-OLT, 50% of patients with post-hepatitis B HCC were alive. The number of acute rejection episodes in patients with recurrent HBV infection and that without recurrent HBV infection was (0.86+/-1.46) time/patient and (1.07+/-0.90) time/patient respectively(P>0.05); the number of moderate acute rejection episodes(RAI>or=score 4) in patients with recurrent HBV infection and that without recurrent HBV infection was (0.29+/-0.49) time/patient and (0.50+/-0.63) time/patient(P>0.05); Incidence of patients with >or=3 episodes of acute rejection in patient with recurrent HBV infection and that without recurrent HBV infection was 14.3% and 10.6%(P>0.05). Furthermore, the number of acute rejection episodes in patients with HCC recurrence or metastasis and that without HCC recurrence or metastasis was (1.12+/-0.93) time/patient and (1.06+/-1.39) time/patient respectively(P>0.05); the number of moderate acute rejection episodes(RAI>or=score 4) in patients with HCC recurrence or metastasis and that without HCC recurrence or metastasis was (0.65+/-0.79) time/patient and (0.65+/-1.06) time/patient respectively(P>0.05); Incidence of patients with >or=3 episodes of acute rejection in patient with HCC recurrence or metastasis and that without HCC recurrence or metastasis was 5.9% and 17.6% respectively(P>0.05). The tumor diameter in patients with HCC recurrence or metastasis was (6.72+/-3.40)cm, however, that in patients without HCC recurrence or metastasis was (3.55+/-2.17)cm(P=0.004 7).The incidence of Portal vein invasion in patients with HCC recurrence or metastasis and that without HCC recurrence or metastasis was 68.75% and 33.3% respectively(P=0.006). CONCLUSION: There was no significant difference among HBV recurrence and acute rejection post-liver transplantation in patients with chronic HBV infection. There was no significant difference between HCC recurrence and acute rejection. The tumor diameter of patients with HCC recurrence or metastasis was significantly greater than that of no HCC recurrence or metastasis. The portal vein invasion of patients with HCC recurrence or metastasis was significantly frequent than that of no HCC recurrence or metastasis.

[Expressions of G2A and OGR1 in peripheral blood cells of patients with hypoxia induced pulmonary hypertension].

OBJECTIVE: To detect the expression changes of proton-sensing receptor G2 accumulation (G2A) and ovarian cancer G protein-coupled receptor 1(OGR1) in human peripheral blood cells in hypoxic pulmonary hypertension patients(HPH). METHODS: Thirty-one patients with HPH were enrolled for HPH group(16 men and 15 women,age:(65.19±5.86) and thirty healthy persons were enrolled for the control group (NC group). The peripheral blood samples were collected and the mRNA expressions of G2A and OGR1 were determined by using real-time fluorescent quantitative PCR. The serum levels of tumor necrosis factor-α (TNF-α) were detected by using enzyme-linked immunosorbent assay (ELISA). RESULTS: PaCO2 was increased significantly in HPH group than that of the NC group (P<0.05). Forced expiratory volume in 1 sencond(FEV1)pro% and FEV1/forced vital capacity(FVC)in HPH group were significant lower than those of the NC group(P<0.05). The expressions of peripheral blood G2A mRNA and TNF-α in HPH group were increased dramatically than those of the NC group(P<0.05). The expressions of OGR1 mRNA in peripheral blood had no difference between HPH group and NC group. The expressions of G2A and TNF-α in HPH group were positively related to pulmonary artery pressure significantly. CONCLUSIONS: The expression of proton-sensing receptor G2A and the level of TNF-α are increased in peripheral blood cells of patients with pulmonary hypertension.The expressions of TNF-α,G2A and pulmonary artery pressure have positive correlation in the HPH group.

The regulatory role of dendritic cells in the immune tolerance.

Immune homeostasis is important for the protection of a host from pathogen aggression, as well as for preventing autoimmunity. Dendritic cells (DCs), the most potent antigen presenting cells, are critical in innate, adaptive immunity and in central tolerance. Recently, their involvement in peripheral tolerance has been shown. Whether DCs induce immunity or tolerance depends on their state of maturation. Different subsets of tolerogenic DCs have been identified in vivo, either in physiological, or pathological conditions, such as tumors, or GVHD. Moreover, tolerogenic DCs can be generated in vitro, by using different culture conditions, such as IL-10 or TGF-beta. In our study, we obtained tolerogenic DCs, by culturing them in the presence of human mesenchymal stem cells (MSCs).

[The expression and significance of urotensin II, nitric oxide, and C-type natriuretic peptide in hypoxia-induced pulmonary hypertension in rats].

OBJECTIVE: To investigate the alteration of plasma and lung tissue homogenate urotensin II (U-II), nitric oxide (NO) and C-type natriuretic peptide (CNP) levels in rats with hypoxia-induced pulmonary hypertension (HPH) and to study the role of these factors and oxygen treatment in the development of HPH. METHODS: Thirty male Wistar rats were randomly divided into three groups: a control group, a group with hypoxia for 14 days and a group with oxygen treatment after hypoxia for 14 days, 10 rats per group. Mean pulmonary arterial pressure (mPAP), mean systematic arterial pressure (mSAP) and right ventriculo hypertrophy index (RVHI) were measured. The plasma levels of U-II, NO, CNP, and the lung tissue homogenate levels of U-II and CNP were measured. The structure of the pulmonary arteries was examined using optical microscope. The microstructure and ultrastructure changes in pulmonary small arteries were examined using electron microscope. RESULTS: The mPAP and RVHI in the hypoxia group [(34.1 +/- 2.8) mm Hg, 0.43 +/- 0.11] were higher than those in the control group [(18.9 +/- 2.0) mm Hg, 0.28 +/- 0.04, all P < 0.01]. The plasma levels of U-II, NO, and CNP [(4.4 +/- 0.9) pg/ml, (20.8 +/- 7.0) micromol/L, (6.6 +/- 1.2) pg/ml], and the lung tissue homogenate levels of U-II and CNP [(6.3 +/- 0.5), (1.89 +/- 0.43) pg/ml] in the hypoxia group were higher than those in the control group [(0.9 +/- 0.4) pg/ml, (13.2 +/- 2.0) micromol/L, (4.0 +/- 0.6) pg/ml, (2.6 +/- 0.5) pg/ml, (0.69 +/- 0.21) pg/ml, respectively, all P < 0.01]. Compared with the hypoxia group, the mPAP, the plasma levels of U-II, NO, and CNP, and the lung tissue homogenate levels of U-II and CNP in the oxygen treatment group [(31.4 +/- 2.0) mm Hg, (2.1 +/- 0.7) pg/ml, (14.8 +/- 1.7) micromol/L, (4.4 +/- 0.7) pg/ml; (3.5 +/- 0.8) pg/ml, (0.74 +/- 0.17) pg/ml, respectively] were significantly decreased (all P < 0.01). The pulmonary vessel morphology changes of the oxygen treatment group were ameliorated. CONCLUSIONS: U-II, NO and CNP are involved in the pathophysiologic process of HPH. Imbalance of these factors may be partially responsible for the development of the disease.

[Monoclonal antibodies against human tumor metastasis suppressor gene-1 TMSG-1: preparation, characterization and application].

OBJECTIVE: In order to clarify the exact molecular weight of tumor metastasis suppressor gene-1 (TMSG-1) protein and its cellular localization, a monoclonal antibody against TMSG-1 was prepared, characterized and applied to evaluate the metastatic potential of human tumors. METHODS: A dominant epitope-TMSG-1(15)-derived from TMSG-1 was synthesized based on Fmoc method, and the hapten was conjugated to Imject Maleimide activated mcKLH as a carrier protein. The antigen preparation was used to immunize BAL B/C mice. Hybridomas were generated and screened by ELISA for specific monoclonal antibodies, which were further characterized by western blotting and immunohistochemical staining. RESULTS: One hybridoma cell line secreting anti-TMSG-1 antibody, designated as C8, was eventually established after primary ELISA screening, followed by rapid limited dilution procedure. It was confirmed that C8 was of IgM isotype. Result of competitive inhibition assay showed that the antibody was TMSG-1 specific. Using this antibody, an expected protein band of about 45,000 (relative molecular mass) was detected in the non-metastatic variants PC(3)-2B4 and PG-LH7 cells by Western blotting, but not in the isogenetic metastatic variants of PC3-1E8 and PG-BE1 cells. Immunohistochemistry using C8 showed a positive staining of cell membrane and cytoplasm of 2B4 and LH7 cells, whereas 1E8 and BE1 cells were non-reactive. Immunostaining using C8 of paraffin sections of 52 breast carcinomas and 41 colon cancers demonstrated a strong positivity in non-metastatic tumors, but none to weakly reactive in metastatic tumors (P < 0.05). CONCLUSION: C8 monoclonal antibody against the synthetic peptide is TMSG-1 specific and is effective for Western blot and immunohistochemistry assays to detect TMSG-1 expression in cancer cells. TMSG-1 protein is about 45 000 (relative molecular mass) at cell membrane and cytoplasm of tumor cells. Expression of TMSG-1 protein correlates well, inversely with the tumor metastatic potential.

[Lung pathology and pathogenesis of severe acute respiratory syndrome: a report of six full autopsies].

OBJECTIVE: Severe acute respiratory syndrome (SARS) is an emerging infectious disease that first manifested in humans in November 2002. The SARS-associated coronavirus (SARS-CoV) has been identified as the causal agent, but the pathology and pathogenesis are still not quite clear. METHODS: Post-mortem lung samples from six patients who died from SARS from April to July 2003 were studied by light and electron microscopy, Masson trichromal staining and immunohistochemistry. Evidence of infection with the SARS-CoV was determined by reverse-transcription PCR (RT-PCR) , serological examination and electron microscopy. RESULTS: Four of six patients had serological and RT-PCR evidence of recent infection of SARS-CoV. Morphologic changes are summarized as follows: (1) Diffuse and bilateral lung consolidation was seen in all patients (6/6) with increasing lung weight. (2) Diffuse alveolar damage was universal (6/6) with hyaline membrane formation (6/6), intra-alveolar edema/hemorrhage (6/6), fibrin deposition (6/6), pneumocyte desquamation (6/6). A marked disruption in the integrity of the alveolar epithelium was confirmed by immunostaining for the epithelial marker AE1/AE3 (6/6). (3) Type II pneumocytes, with mild hyperplasia, atypia, cytomegaly with granular amphophilic cytoplasm and intracytoplasmic lipid accumulation (5/6). (4) Giant cells in the alveoli were seen in five of 6 patients (5/6) , most of which were positive for the epithelial marker AE1/AE3 (5/6), but some cells were positive for the macrophage marker CD68(2/6). (5) A pronounced increase of macrophages were seen in the alveoli and the interstitium of the lung (6/6), which was confirmed by histological study and immunohistochemistry. (6) Haemophagocytosis was present in five of the 6 patients(5/6). (7) Lung fibrosis was seen in five patients(5/6), with alveolar septa and interstitium thickening(5/6), intraalveolar organizing exudates (6/6) and pleura thickening (4/6). Proliferation of collagen was confirmed by Masson trichromal staining, most of which was type III collagen by immunostaining. The formation of distinctive fibroblast/myofibroblast foci was seen in five patients (5/6) by light microscopy and immunochemistry. (8) Squamous metaplasia of bronchial mucosa was seen in five patients(5/6). (9) Thrombi was seen in all patients(6/6). (10) Accompanying infection was present in two patients, one was bacteria, the other was fungus. In addition, electron microscopy revealed viral particles in the cytoplasm of alveolar epithelial cells and endothelial cells corresponding to coronavirus. CONCLUSION: Direct injury of SARS-CoV on alveolar epithelium, prominent macrophage infiltration and distinctive fibroblast/myofibroblast proliferation may play major roles in the pathogenesis of SARS.