Objective To explore the mechanism of vitamin E on delaying skin aging by observ-ing the expression of hyaluronic acid synthetase-2 (HAS-2) in human dermal fibroblasts in vitro. Methods Human skin fibroblasts were cultured in vitro, and these fibroblast cells were then divided into 3 groups: different concentration of vitamin E (0, 0.1 × 10-10, 1 ×109mol/L) was added in the medium in the different group. 24 hours later, the fibroblasts were collected, RNAs extracted, and then amplified by RT-PCR. The PCR product was determined by agarose gel electrophoresis, to analyze the level of HAS-2 mRNA expression. Results RT-PCR showed the lever of HAS-2 mRNA was higher in the low-dose group than the control group, with significant difference (P<0.05) ; the lever of HAS-2 mRNA was higher in the high-dose group than control group, with significant difference (P<0.05). There was no significant difference in the lever of HAS-2 mRNA between the low-dose group and the high-dose group. Conclusions Vitamin E can enhance the expression of hyaluronic acid synthetase-2 mRNA, may increase the synthesis of HAS in skin fibroblasts and increase water content in the skin, so that it might reverse or delay the skin aging.

Objective To investigate the possible mechanisms using different chemicals to improve the skin aging by the effect of estrogen,vitamin E,vitamin C on the transcriptional level of MMP-1 and HAS-2 mRNAs in cultured human skin fibroblast in vitro.Methods The different concentrations of estrogen,vitamin E,vitamin C were put into cultured human skin fibroblasts.After 24 hours,RT-PCR was used to detect the effects of the different chemicals on the transcriptional levels of the HAS-2 and MMP-1 mRNAs in fibroblasts.Results The effects of estrogen and vitamin C on the MMP-1 mRNA had significant difference between the high dose group and blank control group,and between the low dose group and blank control group (P＜0.05) ; the difference between the low dose group and high dose group was not statistically significant (P＞0.05).The effects of vitamin E and vitamin C on Has-2 mRNA also had significant difference between the high dose group and the blank control group,and between the low dose group and blank control group (P＜0.05),but the difference between the low dose group and high dose group was not statistically significant (P＞0.05).Conclusions Estrogen and vitamin C can reduce the transcription of MMP-1 gene,which decreases the degradation of collagen in the skin; vitamin E and vitamin C can increase the transcription of HAS-2 gene,which may possibly increase the synthesis of hyaluronic acid to alleviate the skin aging.

Objective To summarize the effect of surgical treatment and nursing strategies of patients with growth hormone-secreting pituitary adenoma complicated by sleep apnea syndrome. Method Forty-three cases with growth hormone-secreting pituitary adenoma complicated by sleep apnea syndrome underwent surgical treatment in our department , with their history retrospectively analyzed to compare the hormone levels and blood oxygen saturation between pre-and post-operation. Result After the operation, both levels of the hormone and blood oxygen saturation were improved than those before operation; the incidence of sleep apnea syndrome was also lower than that before operation (all P<0.05). Conclusions Surgical treatment is better for patients with growth hormone-secreting pituitary adenoma complicated by sleep apnea syndrome. Reasonable treatment and effective nursing are favorable for the recovery of patients.

Objective To investigate the influences of lysozyme on the mRNA expressions of MMP-1,-12 and lysyl oxidase(LOX)in cultured fibroblasts in vitro.Methods Primarily cultured fibroblasts isolated from human skin were treated with three concentrations(0.1×10~(-8),1×10~(-7)mol/L)of lysozyme followed by another 24-hour cuhure.Subsequently,total RNA was extracted from the fibroblasts and subjected to RT-PCR for the detection of MMP-1,-12 and LOX mRNA.Results There was a significant difference in the mRNA expressions of MMP-1,-12 and LOX among the fibroblasts treated with the three concentrations of lysozyme (F=6.98,4.44,5.24,respectively,all P<0.05).SNK-q test showed that untreated fibroblasts differed signifi-cantly from those treated with lysozyme of 1×10~(-7) mol/L in the mRNA expression of MMP-1 and MMP-12 (P<0.05),and from those treated with iysozyme of 1×10~(-7) mol/L.and 1×10~(-8)mol/L in the mRNA expres-sion of LOX(both P<0.05),whereas no significant difference was ohserved between fibroblasts treated with lysozyme of 1×10~(-8) mol/L and untreated fibrohlasts or those with lysozyme of 1 x 10~(-7)mol/L in the mRNA expression of MMP-1 and MMP-12.or between fibroblasts treated with lysozyme of 1 x 10~(-8)mol/L and those with that of 1×10~(-7) mol/L in the expression of LOX (all P>0.05).Conclusions Lysozyme upregulates the mRNA expression of MMP-1 and MMP-12 but downregulates the mRNA expression of LOX in cultured fibro-blasts in vitro.

AIM: The aim of this work is to investigate the protective effects of Ginsenoside Rb 1(Rb 1) on apoptosis induced by hypoxia /hypoglycemia and reoxygenation in cultured rat hippocampal neurons. METHODS: Apoptosis were measured by flow cytometry; Morphological changes and neuronal necrosis were examined under microscope; The leakage of lactic dehydrogenase(LDH) and the product of nitric oxide(NO) were measured. RESULTS: hypoxia /hypoglycemia cultures for 5 h and reoxygenation induced neuronal apoptosis and necrosis,and significantly increased the leakage of LDH and the product of NO. The effects were enhanced with the extending time of reoxygenation. Rb 1 could significantly decrease the percentage of neuronal apoptosis and necrosis, and reduce the leakage of LDH and the product of NO. CONCLUSION: Rb 1 had an effect of anti-neuronal apoptosis. This effect might be related to the inhibition of the activity of NO synthase and NO production.

Objective To evaluate the clinical efficacy of intra-articular injection of ozone in collagen-induced arthritis (CIA) rats and to assess its effects on serum receptor activator of nuclear factor κB ligand (RANKL) and osteoprotegerin (OPG) levels. Methods Forty weight age malched Wistar rats were randomly divided into normal control group (normal group), the CIA model group (CIA group), ozone (O 3 group), and methotrexate (MTX group). In addition to the normal control group, Freund's complete adjuvant and bovine type Ⅱ collagen were injected to establish the rat model of CIA. After the model was sucessfully developed, double ankle injection concentration ozone group of 40 μg/ml of O3 each 1 ml, 1 times a week, a total of injection for 3 weeks for the experimenal group. MTX group of 0.9 mg/kg was injected 1 times a week for 3 weeks for the MTX group. Degree of foot swelling was measured, and radiographic assessment of arthritis index (AI) score was performed. One week after treatment, angular vein blood was collected for the rats after the intervention, flow multi-factor detection technology was used to test each rat. T test or Wilcoxon rank test was used to compare the difference between groups. Results ① After 3 week administration with O3, dcgree of foot swelling, and AI of the O3 group was reduced significanly than the CIA group during the same period (foot swelling degree: O3 group: (4.21±0.14) ml, CIA group (9.12±0.17) ml, T=64.08, P=0.00; AI O3 group: [(2.97± 0.18) ml, CIA group: 5.76 ±0.13, T=37.24, P=0.00], and X-ray showed joint damage was alleviated. ② The serum level of RANKL in the CIA group was significantly higher than of the normal group [CIA model group 1890.70(797.03, 10571.94)], normal group [74.46(29.21, 95.37), T=43, P=0.005] during the same period; The serum level of RANKL in the O3 group was significantly lower than the CIA group [O3 group 28.09 (14.11, 207.30), CIA group 1890.70 (797.03, 10571.94), T=39, P<0.05]; RANKL level of the Ozone group when compared with MTX group, was not statistically significantly difference (T=52, P>0.05).③Serum RANKL/OPG of the CIA group was significantly higher than that of the normal group during this period, the difference was statistically significant [CIA group 250.68(42.33, 2959.78), normal group 4.32(3.16,5.30), T=36, P<0.01);The serum level of RANKL/OPG in the O3 group was significantly lower than the CIA group [O3 treatment group 5.10 (3.38, 6.64), CIA group 250.68 (42,33, 2959.78), T=36, P<0.01]; There was no statistically significant difference of the serum RANKL/OPG between the O3 group and the MTX group (T=62.5, P>0.05). Conclusion Intra-articular injection of concentration of 40 μg/ml of O3 can reduce RA rat joint swelling degree, which may relate to the mechanism that O3 can lower levels of serum RANKL and RANKL/OPG ratio, reduce osteoclast formation and activation.

Purpose To evaluate the value of real-time contrast-enhanced ultrasound (CEUS) combined with comparative observation in determining the initial time of peripheral lung lesion,and to provide a new method for the diagnosis of peripheral lung disease by CEUS.Materials and Methods Sixty patients with peripheral lung disease diagnosed by chest X-ray or CT examination were examined by CEUS,and the lesions and perfusion of surrounding lung tissue were observed in real time and comparatively.The time of initial enhancement of peripheral lung lesion was determined by real-time comparative observation method,that is,to compare the time of contrast agent to reach the peripheral lung lesion with the time to reach the adjacent gas-containing lung tissue.All lesion specimens were obtained by surgery or ultrasound-guided percutaneous biopsy,and the pathological diagnosis was performed.According to the pathological results,the peripheral lung lesions were classified into the malignant group,the chronic inflammation group and the pneumonia group.Results Sixty cases of peripheral lung lesions were identified as 38 cases of malignant tumors (including 15 cases ofsquamous cell carcinomas,9 cases of adenocarcinoma cell carcinomas,2 cases of large cell carcinomas,9 cases of small cell carcinomas and 3 cases of metastatic carcinomas),14 cases of chronic inflammation lesions (including 7 cases of tuberculomas,3 cases of inflammatory pseudotumors and 4 cases of lung abscess) and 8 cases of pneumonia.The detection rates of pulmonary arterial phase and bronchial artery phase using the real-time comparative observation method were 100％ (60/60) and 85％ (51/60),respectively.The initial time of pulmonary arterial phase and bronchial artery phase were (6.1 ± 0.9) s and (10.5 ± 1.6) s,respectively.The initial time of pulmonary arterial phase was sooner than bronchial artery phase,the difference was statistical significance (P＜0.05).Conclusion The real-time CEUS combined with comparative observation method has important diagnostic value in judging the initial enhancement time of peripheral lung lesion and can provide a new method for the accurate diagnosis of peripheral lung disease,which is worth popularizing.

Objective To observe the effects of intra-articular ozone injection on the secretion of tumor necrosis factor (TNF)-α,Interleukin (IL)-6,IL-17A,and vascular endothelial growth factor (VEGF) in the serum of rats with collagen-induced arthritis (CIA) and explore the therapeutic mechanism of ozone in RA treatment.Methods Thrity-two Wistar rats were randomized into 4 groups,including the ozone groups that receivedintra-articularinjection of 40 μg/ml ozone (O3 group),a blank control group (normal group),a methotrexate (MTX) group (MTX group) anda collagen-induced arthritismodel (CIA group).All the rats,except for those in the blank control group,were subjected to hypodermic injection of bovine collagen Ⅱ and complete Freund's adjuvant to induce CIA.Ozone treatment was administered once weekly for 3 weeks starting at 14 days after the model were established.MTX group were treated with methotrexate 0.9 mg/kg,once a week,a total of three weeks.The swelling degree of the foot were observed,and the serum contents of TNF-oα,IL-6,IL-17A and VEGF were detected.One-way analysis of variance or Kruskal-Wallis test was used to evaluate the experimental data.Results At the end of treatment,the degree of foot swelling was reduced significantly in rats with O3 group compared with that in the CIA group [(4.21±0.14) ml vs (9.12±0.17) ml,t=8.43,P=0.023].The serum concentration of TNF-α,IL-6 and VEGF showed significant difference between the CIA group and the O3 group[91.55(86.55,98.53) pg/ml vs 14.45 (12.55,16.15) pg/ml,x2=6.216,P=0.002;145.08(37.44± 362.82) pg/ml vs 5.84(5.47,15.93) pg/ml,x2=13.136,P=0.004;51.56(46.09,74.10) pg/ml vs.36.22(32.18,41.69) pg/ml,x2=3.732,P=0.002].There was no statistically significant difference between the O3 group and MTX group [14.45(12.55,16.15) pg/ml vs [12.45(11.80,15.60) pg/ml,x2=0.243,P＞0.05;5.84(5.47,15.93) pg/ml vs 7.86(5.25,15.23) pg/ml,x2=0.058,P＞0.05;36.22(32.18±41.69) pg/ml vs 40.17(35.47,50.73) pg/ml,x2=0.516,P＞0.05].The serum concentration of IL-17A showed no significant difference between the normal group,the CIAgroup,the MTX group and the O3 group (F=1.827,P=0.165).Conclusion Intra-articular injecfion of 40 μg/ml ozone can attenuate synovitis in rats with CIA,the mechanism may relate to the inhibition of TNF-oα,IL-6 and VEGF in the serum.

AIM: To study the effect of Jiunaoning Injection on apoptosis and the changes in cytosolic calcium induced by hypoxia/hypoglycemia in SH-SY5Y human neuroblastoma cells. METHODS: The apoptosis and the changes in cytosolic calcium were induced by hypoxia and hypoglycemia .Apoptosis was measured via flow cytometry(FCM), and cytosolic calcium was determined by Fluo-3 fluorometry. RESULTS: The hypoxia/hypoglycemia induced apoptosis and increased the cytosolic calcium in SH-SY5Y cells. Jiunaoning Injection inhibited the apoptosis of the SH-SY5Y cells and decreased the concentration of cytosolic free calcium. CONCLUSION: Hypoxia/hypoglycemia induced apoptosis and increase in cytosolic free calcium in SH-SY5Y cells, which were significantly inhibited by Jiunaoning Injection.

Objective To investigate the feasibility and safety of surgical device closure of doubly committed sub-arterial ventricular septal defect via left sub-axillary.Methods A total of 45 patients diagnosed as doubly committed sub-arterial ventricular septal defect (dcVSD) with transthoracic echocardiography (TTE) and transesophageal echocardiography(TEE) were enrolled from June 2014 to August 2016 in Henan Children Heart Center,Henan Provincial People's Hospital.There were 39 males and 6 females,with the mean age of (2.2 ±2.1) years old(0.5-8.0 years),the body weight (13.8 ± 7.1) kg(7.0-34.1 kg),the defect size (4.5 ± 1.0) mm (3.0-8.0 mm).After general anesthesia,the patients were in supine and evaluated by TEE which indicated whether they were fit to closure.Then,they were turned to the right lateral position while this technique was determined.A vertical incision of 2-3 cm was made between the third and the fifth intercostal space and invasion in thoracic space via fourth intercostal space.Puncture was done at the anterior surface of right ventricular outlet tract to build a delivery tract.The occluder was released and the VSD was occluded under transesophageal echocardiography guidance.Results Forty-one patients had a successful surgical dcVSD closure with asymmetric occluders sized (6.0 ± 1.5) mm(4-10 mm).Among 4 failure cases,2 cases (4.4％) were switched to open-heart surgical repair,1 case (2.2％) due to device related aortic regurgitation,the rest 1 case (2.2％) experienced a dislocation of occluder into pulmonary artery and was converted to surgical repair after retrieve of occluder.Trivial residual shunt was detected in 2 cases (4.4％) postoperatively,a spontaneous closure was observed by 1 month follow-up and 3 months follow-ups,respectively.All the patients were discharged 5 to 8 days after the operation.With a follow-up of (10.4 ±5.0) months [3-24 months],there were no complications such as pericardial effusion,displacement of device,atrioventricular block or new valvular dysfunction.Conclusions Minimally invasive device closure of doubly committed sub-arterial ventricular septal defect via left sub-axillary is a feasible and safe treatment for closure of dcVSD.This technique has advantages of minor wound,less exudation,covert incision,however,long term follow-up is necessary.