[Placenta-derived mesenchymal stem cells with HLA-G positive expression induce Treg in vitro].

OBJECTIVE: To study placenta-derived mesenchymal stem cells with HLA-G (Human Leukocyte Antigen, HLA-G) positive expression induce Treg (regulatory T cell, Treg) in vitro. METHODS: placenta-derived mesenchymal stem cells were separated from neonatal placenta; PEGFP - N1 -HLA-G plasmid was transfected in placenta-derived mesenchymal stem cells by liposome transfection.The cells were divided into 3 groups including control group, PEGFP-N1 group and PEGFP-N1-HLA-G group, 5 complex walls in each group. Expression of HLA-G protein was detected by Western Blotting; after identification of cells, healthy human peripheral blood CD4+ T lymphocytes were cultured with placenta-derived mesenchymal stem cells with HLA-G positive expression, and the ratio of CD4+CD25+Foxp3+Treg in T lymphocytes was accounted. RESULTS: After transfection of PEGFP-N1-HLA-G, the placenta-derived mesenchymal stem cells can express HLA-G protein significantly, compared with the control group and PEGFP - N1 group (P<0.01). After HLA-G positive placenta-derived mesenchymal stem cells and CD4 + T lymphocytes were cultured for 24 h, the ratio of CD4+CD25+Foxp3+Treg in T lymphocytes was (16.41±0.94)%. After HLA - G positive placenta-derived mesenchymal stem cells and CD4+ T lymphocytes were cultured for 48 h, the ratio of CD4+CD25+Foxp3+Treg in T lymphocytes was (16.46±0.59)% significantly, compared with the control group and PEGFP - N1 group (P<0.01). CONCLUSIONS: Placenta-derived mesenchymal stem cells modified by HLA-G gene can effectively induce CD4+CD25+Foxp3+Treg in vitro.

Characteristics and clinical role of bronchoscopy in diagnosis of childhood endobronchial tuberculosis.

BACKGROUND: Endobronchial tuberculosis (EBTB) is the most frequent complication of primary pulmonary tuberculosis (PTB) in children. The aim of the study was to analyze characteristics and clinical role of bronchoscopy in diagnosis of childhood EBTB. METHODS: A retrospective, descriptive study was undertaken in 157 children with EBTB undergone flexible bronchoscopy (FB) between January 2006 and June 2014. RESULTS: The median age of the enrolled patients was 3.4 years, with 73.2% of patients under five years old. The most common subtype was tumorous type (145/157, 92.4%). If only involved bronchus were considered, the common affected sites were right middle lobe bronchus (49/228, 21.5%), left upper lobe bronchus (41/228, 18.0%), right upper lobe bronchus (41/228, 18.0%), right main bronchus (35/228, 15.4%), respectively. Children younger than five years old were at higher risk to have multiple endobronchial lesions (P=0.044), with an odds ratio of 2.313 (95% confidence interval: 1.009-5.299). Before the bronchoscopy, only 16 (10.2%) patients were highly suspected of EBTB, while the others were diagnosed as PTB without EBTB (69.4%), or misdiagnosed as pneumonia or foreign body aspiration (20.4%) on admission. CONCLUSIONS: The patients under five years old are at high risk to progress to EBTB and have multiple endobronchial lesions. The most frequent subtype of EBTB in children is tumorous type. The lesions are seen in the right bronchial system more frequently. FB should be performed to detect the endobronchial lesions in suspected patients as soon as possible.

Temporal Sensing Platform Based on Bipolar Electrode for the Ultrasensitive Detection of Cancer Cells.

We report a bipolar electrode (BPE) sensing platform for the temporal detection of cancer cells. Combining the advantages of anodic dissolution and electrochemiluminescence (ECL), this strategy shows an ultralow detection limit down to 5 cells/cm(2). At the anode working as the reporting pole, Au NPs were assembled through DNA double strand, which served as both catalyzer for the ECL reaction of luminol/H2O2 and seeds for the chemical reduction of Ag, the anodic dissolution probe. The duration of Ag layer dissolution was positively correlated with the amount of Ag but negatively related to the controlled potential and the conductivity of the circuit. Therefore, it was possible to amplify a slight conductivity change through tuning the other two factors. As the formation of Ag@Au completely quenched the ECL emission of luminol, the ECL emission recovery reflected the extent of anodic dissolution. Through monitoring the ECL recovery time before and after the incubation of cells on the cathode, a few number of cells could be quantified due to slight difference of the conductivity. This method shows several merits. First, the combination of anodic dissolution and ECL significantly increases the detection sensitivity of BPE device. In addition, this strategy broadens the application of BPE for the ultrasensitive monitoring of cancer cells, which was applied to investigate the capture efficiencies of antibodies and aptamers toward MCF-7 and A549.

Joint enhancement strategy applied in ECL biosensor based on closed bipolar electrodes for the detection of PSA.

A highly sensitive electrogenerated chemiluminescence (ECL) biosensor was developed on the basis of a closed bipolar electrode (BPE) apparatus for the analysis of prostate specific antigen (PSA). Bipolar modifications bring up two different stages of enhancement on the same electrode. Anodic enhancement was conducted by modifying gold nanoparticles (Au NPs) to catalyze the anodic ECL reaction between luminol and hydrogen peroxide. Cathodic introduction of thionine tagged PSA antibody led to a further pertinently enhancement synchronized with the PSA amount variation, because the existence of thionine greatly increased the rate of electron gains on cathode, leading to the corresponding acceleration of anodic ECL reaction. The more thionine modified target molecules were introduced, the faster luminol was oxidized, the higher faraday current approached, and sensitive quantification was realized in correlation with the responsive ECL intensity differences. The quantification resulted in a good determination range between 0.1pg/mL and 0.1µg/mL. This strategy mainly took advantage of the special structure of closed BPE to realize a simultaneous amplification on both ends of BPE. Moreover, the platform had a potential of providing a multi-functional strategy for the realization of other bio-detections by simply substituting the PSA sandwich structure with other bio-structures.

Feeding methods, sleep arrangement, and infant sleep patterns: a Chinese population-based study.

BACKGROUND: Findings from prior research into the effect of feeding methods on infant sleep are inconsistent. The objectives of this study were to examine infants' sleep patterns by feeding methods and sleep arrangement from birth to eight months old. METHODS: This longitudinal cohort study enrolled 524 pregnant women at 34-41 weeks of gestation and their infants after delivery in 2006 and followed up until eight months postpartum. The study subjects were recruited from nine women and children hospitals in nine cities in China (Beijing, Chongqing, Wuhan, Changsha, Nanning, Xiamen, Xi'an, Jinan, and Hailin). Participating infants were followed up weekly during the first month and monthly from the second to the eighth month after birth. Twenty-four hour sleep diaries recording infants' sleeping and feeding methods were administered based on caregiver's self-report. Multivariable mixed growth curve models were fitted to estimate the effects of feeding methods and sleep arrangement on infants' sleep patterns over time, controlling for maternal and paternal age, maternal and paternal education level, household income, supplementation of complementary food, and infant birth weight and length. RESULTS: Exclusively formula fed infants had the greatest sleep percentage/24 h, followed by exclusively breast milk fed infants and partially breast milk fed infants (P<0.01). Night waking followed a similar pattern. However, the differences in sleep percentage and night waking frequency between exclusively formula and exclusively breast milk fed infants weakened over time as infants developed. In addition, compared to infants with bed-sharing sleep arrangement, those with room sharing sleep arrangement had greater daytime and 24-hour infant sleep percentage, whereas those with sleeping alone sleep arrangement had greater nighttime sleep percentage. CONCLUSIONS: Our data based on caregiver's self-report suggested that partial breastfeeding and bed-sharing may be associated with less sleep in infants. Health care professionals need to work with parents of newborns to develop coping strategies that will help prevent early weaning of breastfeeding.

Taurine drinking attenuates the burden of intestinal adult worms and muscle larvae in mice with Trichinella spiralis infection.

The parasitic nematode Trichinella spiralis can cause trichinellosis, which leads to pathological processes in the intestine and muscle. The intestinal invasion determines the development, subsequent course, and consequences of the disease. Gastrointestinal nematode infection, including with T. spiralis, is accompanied by a rapid and reversible expansion of mucosal mast cell and goblet cell in the intestinal epithelium, which play important roles in the host immune response to parasite and worm expulsion from the intestine. Taurine and its derivatives have anti-infection and anti-inflammatory properties. We investigated whether taurine supplementation in mice could influence the development and pathological processes of infection with T. spiralis. Supplementing 1% taurine in drinking water in mice infected with T. spiralis could alleviate the burden of intestinal adult worms on days 7 and 10 postinfection (all p < 0.01) and the formation of infective muscle larvae in striated muscle during T. spiralis infection (p < 0.01). As compared with T. spiralis infection alone, taurine treatment increased the number of goblet cells on days 7, 10, and 15 (p < 0.01 and p < 0.05) and alleviated intestinal mucosal mast cell hyperplasia on days 10 and 15 (all p < 0.01). So taurine supplementation in drinking water increased infection-induced intestinal goblet cell hyperplasia and ameliorated mucosal mastocytosis. Thus, taurine can ameliorate the pathological processes of trichinellosis and may be of great value for the treatment and prevention of infection with T. spiralis and other gastrointestinal nematodes.

[Diagnostic and therapeutic methods for perioperative children with congenital heart disease with airway stenosis in pediatric intensive care unit].

OBJECTIVE: To explore the diagnostic and therapeutic methods for perioperative children with congenital heart disease (CHD) with airway stenosis in pediatric intensive care unit (PICU). METHOD: Fiberoptic bronchoscopy was used for the diagnosis of 100 CHD cases in PICU who were clinically considered to have possible airway malformation because of complicated difficult-to-control lung infection, atelectasis and failure with the ventilator after surgery from January 2010 to October 2011. Cases who were confirmed to have severe airway stenosis by bronchoscopy and weaning from the ventilator after surgery were treated with balloon expandable stents into the desired position in the bronchoscopy. RESULT: There were 73 cases (73%) of CHD patients with airway abnormalities, including 31 cases of severe stenosis (31%), moderate stenosis in 29 cases (29%), mild stenosis in 13 cases (13%). Nine of the 10 children in whom the mechanical ventilation was hard to be stopped after surgery because of severe airway stenosis were weaned from mechanical ventilation successfully by fiberoptic bronchoscopy, while one case died from primary disease with severe sepsis after the placement of bronchial stents. CONCLUSION: CHD children with difficult-to-control lung infection, atelectasis and failure with ventilator after surgery are often complicated with airway abnormalities. The therapeutic bronchoscopy with airway stent can be used for cases with weaning from the ventilator because of severe airway stenosis.

[Effects of oxidative stress and NF-kappaB levels in peripheral blood mononuclear cells on development of silicosis].

OBJECTIVE: To investigate the change of indicators of oxidative stress in serum and NF-kappaB in peripheral blood mononuclear cells of patients with silicosis, and explore the mechanism of the development of silicosis. METHODS: The subjects were divided into (1) 200 workers exposed to SiO2 for at least 1 years in a foundry served as the dust-exposure group; (2) 130 cases with silicosis (I phase silicosis 64 cases, II phase 46 cases III phase 20 cases) served as the silicosis group; (3) 32 cases with 0+ phase silicosis in the foundry served as the observed group,(4)100 subjects from a hotel served as the control group. The serum including superoxide dismutase (SOD), nitric oxide (NO), serum glutathione peroxidase (GSH-Px), total antioxidant capacity (T-AOC), nitric oxide synthase (NOS), lipid malondialdehyde (MDA) and NF-kappaB protein levels in peripheral blood mononuclear cells were determined, respectively. RESULTS: Compared with the control group, NO levels in dust-exposed group and silicosis group significantly increased, and SOD decreased significantly (P < 0.05 or P < 0.01). Compared with the control group and dust-exposed group, T-AOC, NOS, MDA levels in silicosis group significantly increased (P < 0.05 or P < 0.01). GSH-Px in dust-exposed group and silicosis group were (231.164 +/- 36.484) and (270.469 +/- 39.228)U/ml, respectively which were significantly than that [(223.360 +/- 46.838) U/ml] in control group (P < 0.05 or P < 0.01), and there was significant difference of GSHPx between the silicosis group and the dust-exposed group significantly (P < 0.01) . GSH-Px level [(290.750 +/- 39.129) U/ml] in III phase silicosis group were significantly higher than those [(256.906 +/- 21.41) and (259.594 +/- 34.79) U/ml] in observation group and I phase silicosis group (P < 0.05). NF-kappaB levels [(72.06 +/- 9.12) and (85.25 +/- 11.64) ng/L] in dust-exposed group and silicosis group were significantly higher than that [(59.71 +/- 9.27) ng/L] in control group (P < 0.01), and there was significant difference of between the silicosis group and the dust-exposed group (P < 0.01). There was a positive correlation between serum GSH-Px level and the silicosis stages (r = 0.507, P < 0.01). Also there was a positive correlation between NF-kappaB level and silicosis stages, age, GSH-Px or NO levels (r = 0.376, 0.243, 0.233, 0.221, P < 0.01). CONCLUSION: The imbalance of oxidative and anti-oxidation system and the activation of NF-kappaB are related with the occurrence and development of silicosis. The monitoring of oxidative stress indicators and NF-kappaB is beneficial to the prediction and prognosis assessment of silicosis.

Tricyclo-hex-yl(3,5-dibromo-2-hy-droxy-benzoato-κO)tin(IV).

In the title compound, [Sn(C(6)H(11))(3)(C(7)H(3)Br(2)O(3))], the Sn atom is four-coordinate and possesses a distorted Sn(C(3)O) tetra-hedral geometry, with Sn-C bond lengths in the range 2.132 (6)-2.144 (6) Šand with Sn-O = 2.086 (4) Å. The uncoordinated carboxyl-ate O atom forms a weak contact with the Sn atom, with an Sn⋯O separation of 2.962 (2) Å.

(Benzyl phenyl sulfoxide-κO)dichloridodiphenyl-tin(IV).

The Sn(IV) atom in the title compound, [Sn(C(6)H(5))(2)Cl(2)(C(13)H(12)OS)], displays a distorted C(2)Cl(2)O trigonal-bipyramidal coordination environment, with a mean Sn-C distance of 2.121 (9) Šand with Sn-O = 2.331 (2) Å. The Sn(IV) atom is displaced by 0.169 (2) Šfrom the equatorial C(2)Cl plane towards the direction of the second axially bonded Cl atom.

[Clinical analysis of bronchial foreign bodies in 246 children].

OBJECTIVE: To analyze the characters of bronchial foreign bodies in children and the utilization of bronchoscope in the treatment of bronchial foreign bodies. METHODS: A total of 246 children were diagnosed with bronchial foreign bodies at our hospital during January 2000 until August 2009. Under local mucosal anesthesia, a bronchoscope was inserted through nasal cavity into bronchi. After identifying the site of foreign body, grasping forceps was guided through bronchoscope to remove the foreign body from airway. RESULTS: Among 246 cases, hard nut and skin of melon seed were found (n = 230, 93.5%). The most common site of foreign body was in right lower lobe bronchi (n = 98, 38.9%). The average operative frequency was 1.9 +/- 1.3 and one-time extraction ratio 58.5% (n = 144). The one-time extraction ratio of patients with foreign body obstructed in main bronchi (91.1%), right middle lobe (60.0%) and right lower lobe (55.1%) was higher than others. The operation frequency of using basket grasping forceps (1.4 +/- 0.9) was lower than those using tooth type forceps (2.1 +/- 1.4). And the difference was significant (P = 0.000). CONCLUSION: For bronchial foreign body in pediatric patients, hard nut and skin of melon seed are the most common foreign bodies. The right and left lower lobe bronchi are the predilection site. Foreign body in main bronchus is the easiest to be removed by grasping forceps. For massive foreign bodies, basket grasping forceps fares better than tooth grasping forceps.

(E)-2-[(2-Hydroxy-ethyl)iminiometh-yl]-6-methoxy-phenolate.

The title Schiff base compound, C(10)H(13)NO(3), obtained by the reaction of 2-hydr-oxy-3-methoxy-benzaldehyde and 2-amino-ethanol in methanol solution, crystallizes in a zwitterionic form, in which the mol-ecule adopts a trans configuration about the central C=N bond. An intra-molecular N-H⋯O hydrogen bond occurs. In the crystal structure, mol-ecules are linked into chains by inter-molecular O-H⋯O hydrogen bonding.

[Measurement of pulmonary function with body plethysmograph in infants].

OBJECTIVE: To observe the alterations of pulmonary function in infants with respiratory diseases. METHODS: A total of 900 infants with respiratory diseases were recruited and pulmonary function measured in 30 healthy infants. The tests were performed in the sleeping infants with sedation. Tidal breathing flow-volume (TBFV) loops were recorded when infants were breathing quietly. Passive flow-volume technique was used to obtain static respiratory system compliance and resistance. Functional residual capacity was measured by body plethysmograph. RESULTS: The TBFV loop showed proximate round or oval curve in healthy infants. The shape and parameters of TBFV loop had significant differences in infants with respiratory diseases as compared with healthy controls. The TBFV loop displayed a concave expiratory curve and ratio of time to reach tidal peak flow to total expiratory time, the expiratory volume till peak flow divided by the total expiratory volume significantly decreased in infants with small airway obstruction. The expiratory or inspiratory curve showed a plateau and the ratio of mid-expiratory to mid-inspiratory flow was less than 60% or over 150% in infants with upper airway obstruction. The TBFV loop turned narrow and lung volume decreased in infants with restrictive diseases. CONCLUSION: The TBFV loop show proximate round or oval curve in healthy infants. Pulmonary function has significant differences between healthy controls and infants with respiratory diseases. Pulmonary function test is useful in the assessment of respiratory diseases.

Proteomic analysis of human NK-92 cells after NK cell-mediated cytotoxicity against K562 cells.

To better understand the natural killer (NK) cell cytotoxicity mechanism at the proteome level, we comparatively analyzed the proteome of the human NK-92 cells which participate in NK cell-mediated cytotoxicity assay and that of control cells. Soluble proteins were separated by two-dimensional gel electrophoresis (2-DE), 75 protein spots were found to be reproducibly differentially expressed between control and cytotoxic human NK-92 cells. A total of 60 different proteins were unequivocally identified by MALDI-TOF MS coupled with database interrogation; 37 proteins were up-regulated, whereas 23 proteins were down-regulated. Western blotting analysis of heat shock protein 60 (HSP60) and cathepsin W verified their proteome results. Some of identified proteins are involved in NK-92 cytotoxicity, which is consistent with the literature. In addition, we modeled the pathway networks between differentially expressed proteins and cellular processes of secretion and exocytosis through PathwayStudio software. The results of this study help to provide insight into the molecular mechanism of NK cell cytotoxicity.

[Study on prevalence and risk factors of sleep disorder among Chinese children aged 0 to 23 months in city].

OBJECTIVE: To investigate the prevalence and main influences on sleep disorder among Chinese children aged 0 to 23 months, as to providing scientific interventions for infant sleep disorder. METHODS: All 7601 children under two years old were selected by stratifying samples from twelve cities in China. The objects' parents were surveyed with questionnaire. All data were analyzed with SPSS statistical software. RESULTS: The total incidence of sleep disorders at 0 to 23 months was 21.94%. The main problems were difficulty falling asleep, nighttime waking and snoring. Feeding manner, sleep environment, sleep-associated habits and medical conditions were all influences on infant's sleep disorder. CONCLUSIONS: Enhancing sleep health education to change parents' nurturing modes should be an important role in preventing infant sleep disorders.

Soluble expression of active human beta-defensin-3 in Escherichia coli and its effects on the growth of host cells.

BACKGROUND: Human beta-defensin-3 (HBD(3)) is an epithelial peptide that has been demonstrated to have a salt-insensitive broad spectrum of potent antimicrobial activity. Expressing antimicrobial peptides in Escherichia coli (E. coli) is very difficult for it can result in death of the bacterial host cells. Our aim was to establish a prokaryotic system expressing soluble HBD(3) protein and demonstrate the antimicrobial activity of the expressed protein. We then studied whether the host cells would activate the suicide pathways. METHODS: We first cloned the complementary DNA coding for the mature chain of HBD(3), inserted it into the vector PGEX-KG then transformed E. coli BL21 (DE3) with the appropriate recombinant plasmid. After induction with 0.5 mmol/L isopropyl-1-thio-beta-D-galactopyranoside (IPTG) the transformed E. coli produced a recombinant glutathione S-transferase and HBD(3) (GST-HBD(3)) fusion protein. The fusion protein was treated with thrombin to produce pure HBD(3) protein then the antimicrobial activity of HBD(3) was evaluated in a liquid microdilution assay. RESULTS: The fusion protein GST-HBD(3) was efficiently cleaved by thrombin and yielded HBD(3) that had anti-staphylococcus aureus activity with a minimal inhibitory concentration level of 12.5 microg/ml. The E. coli strain expressing the recombinant protein did not grow slower than the empty vector strain. CONCLUSION: Active HBD(3) in E. coli by expressing the recombinant protein GST-HBD(3) could be produced, and suicide did not occur in the E. coli strain expressing the recombinant protein.

[Galectin-7 is associated with bronchial epithelial cell apoptosis in asthmatic children].

OBJECTIVE: It is supposed that bronchial epithelial cells responses to the environmental stimuli are different between asthmatic and non-asthmatic individuals, which contribute to the pathogenesis of asthma. These different responses produce different mediators. If differential gene expressions are found in bronchial epithelial cells of asthmatic and non-asthmatic individuals after the same stimuli in vitro, and these genes are overexpressed in asthmatic children in vivo, then it is concluded that these genes may be associated with asthma. Therefore the authors analyzed the differential gene expressions in the bronchial epithelium cells of asthmatic and non-asthmatic children after RSV infection in vitro. Among these genes, Galectine-7 (lectin, galactoside-binding, soluble, 7, Galectin-7) was 8 times up-regulated in asthmatic children. Galectine-7 was associated with skin keratinocyte apoptosis. The authors hypothesized that Galectin-7 may also be associated with bronchial epithelial cell apoptosis in asthmatic children. The aim of this study was to understand the role of Galectine-7 in bronchial epithelial cell apoptosis in asthma. METHODS: The bronchial mucosae of one asthmatic child and one non-asthmatic child were obtained by biopsy and cultured in vitro. The bronchial epithelial cells were infected by RSV. The differential gene expressions were analyzed with micro array. Among those differentially expressed genes, Galectin-7 was 8 times up-regulated in asthmatic children. The bronchial mucosae from 10 asthmatic children and 17 non-asthma children were investigated for cell DNA break, Galectine-7 and mRNA expression, Caspase-3 expression by TUNEL, hybridization in situ and immunochemistry. Image analysis was used for quantitative assessment. RESULTS: Galectine-7 gene was 8 times up-regulated in bronchial epithelial cells from asthmatic children after RSV infection in vitro. Galectin-7 and mRNA were overexpressed in bronchial epithelial cells in asthma in vivo. Bronchial epithelial cell apoptosis increased in asthma in vivo. CONCLUSION: Galectin-7 may be associated with bronchial epithelial cell apoptosis in asthma.