Objective To investigate the effects of cobalt chloride(COCl2)induced rat alveolar epithelial cell apoptosis and the significance of related apoptosis protein hypoxia inducible factor-lalpha(HIF-1 a)expression. Methods Alveolar epithelial cells were cultured in vitro and divided into four groups with 6 samples in each.Hypoxia 4 h group:added 500μmol/L COCl2 for 4 hours inducing alveolar epithelial cell hypoxia;hypoxia 24 h group:co-cultured with COCl2 for 24 hours;hypoxia 48 h group:co-cultured with CoCl2 for 48 hours;control group:no CoCl2 added.The apoptosis rates were detected by fluorescent microscope and flow cytometry.The apoptosis morphological changes of the cell were observed by transmission electron microscopy.The protein expression of HIF-la was detected bv Western Blot. Results (1)Fluorescent microscope results showed that apoptosis rate in hypoxia 4 h、24 h、48 h group[(5.83±0.76)%,(15.00±3.28)%and(51.50±3.00)%],were higher than control group[(1.50±0.50)%](P＜0.05).(2)Flow cytometry analysis reached the same results as with fluorescent microscope.(3)Typical morphological changes of apoptosis were observed in hypoxia 24 h group.(4)HIF-1α expression in hypoxia 4 h、24 h and 48 h groups[0.69±0.035,1.02±0.044 and 0.71±0.046],were higher than control group(0.21±0.026)(P＜0.01).(5)Positive correlations were found between relative amount of HIF-1αprotein and apoptosis rate by fluorescent microscope and flow cytometry analysis(r=0.484,P=0.016;r=0.713,P=0.009).Conclusions Hypoxia could induce apoptosis of alveolar epithelial cells,which may participate in the pathogenesis of hypoxia-induced lung injury.

Abscess ; pathology ; Adenoma ; metabolism ; pathology ; Adenoma, Sweat Gland ; metabolism ; pathology ; Biomarkers ; metabolism ; Breast Diseases ; pathology ; Breast Neoplasms ; metabolism ; pathology ; Female ; Fistula ; pathology ; Humans ; Keratin-7 ; metabolism ; Keratins ; metabolism ; Mucin-1 ; metabolism ; Nipples ; pathology ; Paget's Disease, Mammary ; metabolism ; pathology ; Receptor, ErbB-2 ; metabolism ; Sweat Gland Neoplasms ; metabolism ; pathology

Objective To investigate the use of feeding support and influencing factors of oral feeding among premature infants. Methods Totally 103 premature infants with oral feeding problems hospitalized during June to December 2014 were enrolled. The postmenstrual age and feeding support were observed at the time of initiation oral feeding and full oral feeding. Moreover, the progress and performance of oral feeding, the daily increase in weight, the time of body weight regain, the time of assisted ventilation and length of hospital stay and so on were calculated. Results There were 96 premature infants using feeding support at the time of initiation oral feeding and 58 premature infants at the time of full oral feeding. Logistic regression analysis showed that birth gestational age(OR=2.195, P<0.01), birth weight(OR=1.003, P<0.01), severity of illness(OR=0.121, P<0.01) and feeding tolerance(OR=0.007, P<0.01)were important factors of oral feeding among premature infants. Conclusions The premature infants with small gestational age at birth, low birth weight, severity of illness, and feeding intolerance are high risk populations of oral feeding. Nurses should take timely risk assessment and interventions to reduce the occurrence of stopping oral feeding.

Objective To examine expression of PTEN gene in ovarian cancer cisplatin-sensitive cell line OV2008 cells and cisplatin-resistant cell line C13K cells,and evaluate the effect of wild-type PTEN gene on reversing cisplatin-resistance of C13K cells and underlying mechanisms.Methods The expression of PTEN mRNA and protein in OV2008 and C13K cells were detected by semi-quantitative RT-PCR and western blot.Recombinant eukaryotic expression plasmid containing human wild-type PTEN gene was transfected into C13K cells by lipofectamine 2000.The expression of PTEN mRNA was monitored by RT- PCR and the expression of PTEN,protein kinase B(AKT),phospho-AKT(p-AKT)protein were analyzed by western blot in PTEN transfected and untransfected C13K cells.Proliferation and chemosensitivity of cells to cisplatin were measured by methyl thiazolyl tetrazolium(MTT),and cell apoptosis was detected by flow cytometry after treatment with cisplatin.Results(1)The expression of PTEN mRNA and protein(1.02 ?0.05,1.02?0.07)in OV2008 cells were significantly higher than those in C13K cells,which were 0.45 ?0.03 and 0.55?0.03 respectively(P

Neurotensin receptor-1 (NTR1), which can stimulate the intracellular cascade signal pathway, belongs to the large superfamily of G-protein coupled receptors. NTR1 is related to the occurrence and development of several kinds of diseases. In order to screen the inhibitors for the cancers associated with NTR1 protein, we established a CHO (Chinese hamster ovary) cell line in which human neurotensin receptor-1 was highly expressed. The method is to construct the recombinant plasmid which was lysed with the hNTR1 gene and transfect it into CHO cells. After selected with G418, the cell line was evaluated by Western blotting analysis and calcium flux assays. Through the calcium flux assays on FlexStation 3, we got the EC50 value of neurotensin peptide which is the natural NTR1 agonist, and the IC 50 value of SR48692 which is the known NTR1 antagonist. The established human CHO/NTR1 cell line can be used to study the profile of NTR1 biological activity and further screen of NTR1 antagonists and agonists.
Animals ; CHO Cells ; Calcium Signaling ; Cricetinae ; Cricetulus ; Humans ; Pyrazoles ; pharmacology ; Quinolines ; pharmacology ; Receptors, Neurotensin ; antagonists & inhibitors ; genetics ; metabolism ; Transfection

Objective To explore the clinical characteristics of neonatal congenital cystic lung lesions. Methods Between January 2008 and June 2014,total 28 cases diagnosed congenital cystic lung lesions in neonatal center of Guangzhou Women and Children's Hospital were collected.The clinical data were analyzed including manifestations,lesion characteristics,imaging,diagnosis,treatment and prognosis,and the related literature were reviewed.Results There were 20 male and 8 female,16 cases of lobar emphysema,7 cases of pulmonary seques-tration,4 cases of congenital cystic adenomatoid malformation and 1 case of bronchogenic cyst.Main symptoms were dyspnea(78.57%),cyanosis (39.29%),wheezing cough (17.86%),feeding difficulties (14.29%),fever (10.71%),asymptomatic(21.43%).Two cases combined with congenital heart disease,4 cases combined with other malformations(such as diaphragmatic hernia,laryngeal stridor,funnel chest,polycystic kidney).There were lesions in chest CT image,4 cases underwent lobectomy,other cases underwent conservative treatment.Seventeen cases got better and discharge,3 cases dead.Conclusion The most common type of congenital cystic lung lesions in neonate is lobar emphysema.Main manifestations are dyspnea,cyanosis.If we suspect congenital cyst-ic lung lesions,we should do chest CT or MRI scan soon ,also do echocardiography and ultrasound to exclude other malformations.The treatment effect is satisfactory,and the outcome need long time to follow up.

BACKGROUND:Corneal lymphangiogenesis is beneficial to the transport of corneal antigenic materials, and accelerates the process of antigen presentation, thereby playing an important role in corneal immunity. However, due to the paral el outgrowth of corneal blood and lymphatic vessels in transplanted corneas, it is often difficult to accurately evaluate the role of corneal lymphatic vessels in allograft rejection. OBJECTIVE:To explore the development of corneal lymphangiogenesis and angiogenesis in transplanted rat corneas after the blockade of vascular endothelial growth factor C (VEGF-C). METHODS:130 rats used to establish corneal al ogenic transplantation models were equally randomized into two groups:the anti-VEGF-C group and the control group. VEGF-C was blocked in the anti-VEGF-C group by intraperitoneal injection of neutralizing monoclonal anti-VEGF-C antibody every other day for 2 consecutive weeks. Meanwhile, rats in control groups received intraperitoneal injections of saline. Corneal angiogenesis and lymphangiogenesis were characterized using whole mount immunofluorescence, and the immune rejection of the grafts was evaluated by scoring the rejection index (RI). In addition, the expression of VEGF-C was examined by real-time PCR. The relationship of corneal lymphangiogenesis and angiogenesis to RI in transplanted corneas was also characterized. RESULTS AND CONCLUSION:VEGF-C expression was markedly downregulated after VEGF-C blockade. Corneal lymphangiogenesis developed in parallel with corneal angiogenesis in the control group. While there was a mild reduction in blood vessel area (BVA) and a significant decrease in lymphatic vessel area (LVA) in the anti-VEGF-C group (P<0.05). In addition, RI was positively correlated with BVA (P<0.05) and LVA (P<0.05) in the control group. However, although RI was significantly correlated with BVA (P<0.05) in the anti-VEGF-C group, the correlation between RI and LVA was not statistically significant (P>0.05). the graft survival time in the anti-VEGF-C group was significantly increased compared with that in the control group (P<0.05). Our results show that the outgrowth of lymphatic vessels is separated from that of blood vessels in transplanted corneas by blocking VEGF-C. The blockade of VEGF-C has a significant role in preventing corneal lymphangiogenesis in corneal beds, which results in higher al ograft survival rates.

Forty batches of Lonicerae Japonica Fse i collected extensively and prepared as the test solution. Their chromatographic fingerprints and anti-influenza virus IC50 value (half maximal inhibitory concentration) were determined respectively. Then Unscrambler software was used, and spectrum-efficient correlation analysis was done for chromatographic fingerprints data and IC50 data by partial least squares regression method, to establish spectrum-efficient correlation model for anti-influenza virus of Lonicerae Japonicae Flos. Then the other 10 batches of Lonicerae Japonicae Flos were used to verify the model and explore the adaptability of this spectrum-efficient correlation model based on partial least squares regression method. The mathematical model obtained R2 of 0.969489 and RM-SEC of 0.070691 for calibration set; R2 of 0.959042 and RMSECV of 0.084005 for cross validation set. The verification experiment results showed that the relative error between the predicted values and measured values was within 10% in all 10 hatches, and within 5% in 80% of them. The results showed that the established spectrum-efficient correlation model could be used to evaluate the biological activity of anti-influenza virus of Lonicerae Japonicae Flos by determining its HPLC fingerprints.
Antiviral Agents ; analysis ; pharmacology ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; analysis ; pharmacology ; Flowers ; chemistry ; Least-Squares Analysis ; Lonicera ; chemistry ; Molecular Structure ; Orthomyxoviridae ; drug effects

AIM: To screen TIGR/myocilin gene (MYOC) mutation in high myopic Chinese children with family history.METHODS: Gene sequencing was performed in exon 3 of the TIGR gene in high myopic Chinese Children. The coding sequence of TIGR exon 3 was screened by capillary electrophoresis sequencing. The sequence alterations were analyzed by bioinformatics.RESULTS: TIGR gene mutation was not found in high myopic patients and normal controls group.CONCLUSION: No identified gene mutation is found in TIGR gene in high myopic Chinese children.

OBJECTIVETo evaluate the role of brain magnetic resonance imaging (MRI) and tumor markers in the cerebral spinal fluid (CSF) and serum in the diagnosis and treatment of intracranial germinoma in children.

METHODSTotally 5 children (3 girls and 2 boys) who were treated in our hospital between January 2009 and December 2010 due to central diabetes insipidus. All patients received contrast-enhanced brain MRI at presentation and during each follow-up: meanwhile, their anterior pituitary hormones and tumor markers including human chorionic gonadotropin (hCG) and alpha fetoprotein (AFP) were also determined.

RESULTSThree patients presented without prior evaluation, and two patients were referred to our hospital due to exaggerated disease of unknown cause. Their ages at presentation ranged from 8 years to 12 years 1 month, and the duration of symptoms at presentation was between 1 month to 78 months. All of them had polyuria and polydipsia at presentation. Except one child, the other 4 patients had growth retardation and failure in initiation of puberty. Although the growth rate and puberty development were normal during the 2-year follow-up for the excepted child, all child experienced anterior pituitary hypofunction and an increased concentration of plasma prolactin after the lesion became enlarged. Three patients had cerebral hernia, which presented in 18, 24, and 78 months, respectively. In three patients, brain MRI at presentation showed isolated pituitary stalk thickening, which further developed into massive tumor in the hypothalamus pituitary region 18-22 months later; in the remaining two patients, large brain tumor was found via MRI at their first presentations. In all five patients, the posterior pituitary gland (bright spot) disappeared on T1-weighted MRI images. CSF hCG elevated in all five patients, and serum hCG increased in four patients; the level of hCG varied with the mass size of tumor. Serum and CSF AFP increased in only one patient.

CONCLUSIONSPatients with idiopathic central diabetes insipidus must be closely followed to identify the etiology, especially when anterior pituitary hormone deficiencies are detected. For patients with normal brain MRI results or simply isolated pituitary stalk thickening at presentation, the changes of serial contrast-enhanced brain MRI should be observed during follow-up to ensure the early detection of an evolving occult hypothalamic-stalk lesion. Determination of CSF hCG at the first presentation may be useful, because an increased CSF level of hCG precedes MRI abnormalities.

Biomarkers, Tumor ; analysis ; blood ; cerebrospinal fluid ; Brain Neoplasms ; blood ; cerebrospinal fluid ; diagnosis ; Child ; Female ; Germinoma ; blood ; cerebrospinal fluid ; diagnosis ; Humans ; Magnetic Resonance Imaging ; Male ; Retrospective Studies