Objective To investigate p16 promoter methylation in premature rats with chronic lung disease induced by hyperoxia. Methods Eighty premature Wistar rats were randomly divided into two groups: hyperoxia group (fraction of inspiratory oxygen) 0. 90 and control group (fraction of inspiratory oxygen 0. 21), 40 rats for each group. Semi-nested methylation specific polymerase chain reaction and methylation specific polymerase chain reaction were applied respectively to detect p16 promoter methylation in lung tissues. Additionally, p16 mRNA and protein expressions in lung tissue were detected by reverse transcription- polymerase chain reaction, Western blot and immunohistochemistry method. Results The methylation was not found in control group by seminested methylation specific polymerase chain reaction and methylation specific polymerase chain reaction, while was found in different aged rats of the hyperoxia group. The methylation detection rate was higher by using the semi-nested methylation-specific polymerase chain reaction (52.5%, 21/40) than that by methylation specific polymerase chain reaction (42.5%, 17/40) in the hyperoxia group,but there was no statistically significant difference between the two methods. The p16 mRNA in the hyperoxia group were significantly lower than in the control group at day 7, 14 and 21(1.73 ± 0.40 vs 2.11±0. 37,1.29±0. 19 vs 1.60±0. 27,0. 95±0.25 vs 1.72±0. 34, t=2.19, 2.95 and 10. 43,P＜0. 05). The p16 protein expressions by western blot in the hyperoxia group were significantly lower than in the control group at day 7, 14 and 21 also (88. 1±8. 7 vs 95.0±4.1,65.7±4.5 vs 83. 5±13.6 and 50.4±4.9 vs 86.7±11.9, t=2.27,3.95 and 13.40,P＜0.05). The expression of p16 mRNA (1.06±0.61) and protein (62.32±25.65) in lung tissues of rats with methylation was lower than that without methylation (1.63±0.62 and 94.93±22.21, respectively) (t=2.95, OR=0. 86;t=4.28, OR=0. 85,P＜0.01, respectively). Conclusions Exposure to hyperoxia might induce p16 promoter methylation in lung tissues in premature rats. Methylation risk increases as exposure time extends. p16 promoter methylation induced by hyperoxia might participate in the mechanism of lowering p16 mRNA and protein expression, but might not result in p16 gene silence.

Adrenal Gland Diseases ; diagnosis ; pathology ; therapy ; Female ; Hemorrhage ; diagnosis ; pathology ; therapy ; Humans ; Infant, Newborn ; Magnetic Resonance Imaging ; Male ; Prognosis ; Tomography, X-Ray Computed

AIM:To study the clinical effect of silica gel drainage tube combined with lomefloxacin hydrochloride eye ophthalmic gel in the treatment of lacrimal duct obstruction.METHODS:Totally 86 cases (138 eyes) of lacrimal duct embolism treated in our hospital from February to December 2015 were divided into 43 cases (68 eyes) as control group and 43 cases (70 eyes) of the observation group according to whether they were treated with lomefloxacin hydrochloride eye ophthalmic gel.Patients in the control group were treated with silica gel drainage, while the observation group was treated with lomefloxacin hydrochloride eye ophthalmic gel on the basis of the control group.The total effective rate, serum hypersensitivity C-reactive protein (h-CRP) level, complications and recurrence rate were observed and compared between the two groups.RESULTS:The total effective rate in the patients with obstructive nasolacrimal duct, common lacrimal duct obstruction, and lacrimal duct obstruction in observation group were 95.2%, 100.0%, 96.7%, higher than those in control group (P<0.05).There was no significant difference between the two groups in preoperative serum h-CRP levels (P>0.05).The serum levels of h-CRP in patients in the observation group at 3d and 7d after operation respectively were 2.40±0.84g/mL, 1.94±0.84g/mL, lower than those of control group at the same time (P<0.05).The complication rate of the observation group was 1.4%, which was lower than that of the control group (P<0.05).Follow up for 6-18mo, the recurrence rate was 11.4% in the observation group, which was significantly lower than the control group (P<0.05).CONCLUSION:The combination of silica gel drainage tube and lomefloxacin hydrochloride eye ophthalmic gel in the treatment of obstruction of lacrimal duct has good clinical efficacy, low inflammatory reaction, low complication rate and low recurrence rate.

Objective: To explore the burden of spouse caregivers of patients with dementia at home and factors having influence.Methods:Forty-two patients with dementia were collected from psychiatric out-clinic consecutively. Trained investigators did home visit to collect data on severity of the disease (CDR, MMSE), pathological behavior (BEHAVE-AD), activity of daily living (ADCS-ADL) and burdens of their spouse as caregiver (CBI). The caregivers' anxiety, depression (Zung's Anxiety Scale and Depression Scale) and knowledge about dementia were also collected.Results:The caregivers felt big burden (BDI total score: 30.7?15.1). Nearly half of them (42.9%) were in depression and one quarter (23.8%) were anxious. They were lack of knowledge about dementia and unskillful in care-giving. 26.2% caregivers thought dementia is curable or equal to normal aging, 57.1% of them did not believe it is an improving disease. The burden of caregivers was correlated positively with pathological behavior of patients (r=0.40, P

Objective To establish a LC-MS analytical method for determination of N,N-dimethylaniline which is genotoxic impurity in quetiapine fumarate.Methods The method was achieved by an Waters ACQUITY UPLC CSHTM PhenylHexyl(2.1 mm× 100 mm,1.7 μm) utilizing a mobile phase of buffer-methanol(900∶ 100) (A)-acetonitrile(B) with gradient elution at the flow rate of 0.4 mL·min-1.The temperature of column was set at 50 ℃;The DIONEX Ultimate 3000 HPLC-AB Science 4000 QTrap Tripling Four bar LC-MS to detect N,N-Dimethylaniline (ESI source,in MRM positive mode).Results Standard curve was linear in the range of 0.4-8.0 ng(r =0.999 3);The limit of detection was 0.2 ng;The limit of quantification of N,N-dimethylaniline was 0.4 ng,respectively.The average recovery of N,N-dimethylaniline was 103.3 ％;RSD was 4.3％ (n =9),respectively.Conclusion The method is convenient and sensitive for the determination of N,N-dimethylaniline in quetiapine fumarate.

Using hitochemical techniques showing cholinesterase and catecholamines,weinvestigated the changes of these neurotransmitters in the sympathetic nervous centers(posterior nucleus and lateral area of hypothalamus,as well as intermediolateralnucleus of spinal cord)of acupunctured rats,and compared with those in the cor-responding nuclei and area in normal controls.A total of 109 cases were performed,among which 73 cases were of acupunctured and 36 cases were of controls.Theresults obtained were as follows:(1)It was shown that the number of cases withstronger activity of acetylcholinesterase in sympathetic nervous centers of acupunc-tured group was higher than that of controls.The more obviously elevated in painthreshold after acupuncture,the higher number of cases with stronger ACHE enzy-matic activity in the sympathetic nervous centers was evident.(2)The number ofcases with stronger intensity of NA fluorescences within the adrenergic fiber's terminalsof sympathetic nervous centers was found to be higher in acupunctured group thanthat of control group.On the basis of the results obtained,the possible relationship between the effectsof acupuncture and the changes of neurotransmitters in sympathetic nervous centerswas discussed.We considered that the sympathetic nervous centers might have beeninduced into a relative inhibition state following acupuncture,and this state is ingood coincidence with the increase of NA in adrenergic fiber's terminals.

AIM: To establish the quality control for Lidanzhitong Tablet (Radix Paeoniae Rubra,Fructus Aurantii,Rhizoma Corydalis,etc.). METHODS:Radix Paeoniae Rubra, Fructus Aurantii and Rhizoma Corydalis were identified by TLC, and the content of paeoniflorin in Radix Paeoniae Rubra was determined by HPLC. RESULTS: Paeoniflorin showed a good linear relationship at a range of 0.597-4.997 ?g, r=0.999 9. The average recovery was 99.06%, and RSD was 3.4%. CONCLUSION: The method is accurate and reliable, and can be used to control the quality of this preparation effectively。

Objective To compare the curative effects between domestic and imported monosialotetrahexosyl ganglioside sodium (GM 1) on neonatal hypoxic ischemic encephalopathy(HIE).Methods 104 neonates with HIE were randomly divided into Shenjie group(domestic GM 1,53 cases )and Pharma group (imported GM 1,51 cases).At the basic of conventional therapy,the two groups received domestic or imported GM 1 20 mg /d intravenous infusion for 7～ 28 d,rspectively.The curative effects and adverse reaction were observed.Results The total effective rates in Shenjie group and Pharma group were 98.11% and 88.24%,respectively.There was no significant difference between the two groups.While,there was no obvious adverse reaction in the two groups.Conclusion Both the domestic and imported GM 1 have the same good curative effects and safety on neonatal HIE.

Objective To investigate the functional role of mur in e hepatocyte growth factor (HGF) in inducing in vitro the mouse embryonic st em cells (ESC) to differentiate unidirectionally into hepatocyte. Method s Both spontaneous and HGF induced differentiations of the primordial c ell elusters formed by suspended culture in vitro of ESC were daily observed with microscope for 5-7 days. After 4 weeks of culture, the cells were stained with HE and glycogen staining, their morphology were observed, The synthesis of urea nitrogen and triglycerides in the culture medium, the expression of myocard ial MHC, albumin, AFP and CK18, and the indocyanine green (ICG) staining were a lso detected. Results It was hard to control the spontaneous di fferentiation of ESC. HGF could promote the differentiation of ESC into endoderm , and more likely into myocardium. HGF could also induce the expression o f albumin, AFP and CK18, and positive staining of ICG and FDA. Conclusio ns HGF may induce the differentiation of ESC into hepatocyte, but the f unctional role is limited. It implies that a comprehensive effort of multiple fa ctors might be needed in inducing the hepatocyte differentiation.

Objective To observe the changes of neutrophil gelatinase associated lipocalin,NGAL after endoplasmic reticulum stress by thapsigargin in renal tubular epithelial cells.Methods The renal tubular epithelial cells were treated with thapsigargin to establish the model of endoplasmic reticulum stress (ER group).CHOP and GRP78 expression were detected by Western-blot to confirm endoplasmic reticulum stress in renal tubular epithelial cells by thapsigargin.NGAL expression were detected by Western-blot in ER group and control group(normal renal tubular epithelial cells).Results After treated with 2.5 μmol/L and 5 μmol/L thapsigargin for 4 hours and 8 hours respectively,endoplasmic reticulum stress in the renal tubular epithelial cells were significantly increased comparing with control group (0.585 ± 0.045 to 0.523 ± 0.030;0.785 ± 0.049 to 0.728 ± 0.064),which was approved by the increased expression of CHOP and GRP78 (P ＜ 0.05).The expression of NGAL was significantly increased in ER group (0.567 ± 0.024 to 0.826 ± 0.057,P ＜ 0.05) and it was corresponded with the changes of CHOP and GRP78,which increased significantly with the severity of endoplasmic reticulum stress.Conclusion Endoplasmic reticulum stress can be induced by thapsigargin in renal tubular epithelial cells.The expression of CHOP,GRP78 and NGAL were significantly increased correspondingly with the severity of endoplasmic reticulum stress.It is speculated that the expression of NGAL may be related with endoplasmic reticulum stress in tubular epithelial cells.