Objective:To discuss the damage effect of vesicular stomatitis virus(VSV)on the vascular endothelial(VE)barrier,and to clarify its mechanism.Methods:The canine kidney cells were used to amplify VSV.The half tissue culture infective dose(TCID50)of VSV was determined using mouse brain endothelial tumor bEnd.3 cells,and subsequent experiment was conducted using 300 times the TCID50.The bEnd.3 cells were divided into infection 0 h group,infection 4 h group,infection 8 h group,and infection 12 h group for VE barrier damage experiments due to VSV infection.The bEnd.3 cells were also divided into control group,infection group,and correction group for experiments to inhibit the VSV replication and restore the VE barrier.The bEnd.3 cells were inoculated into Transwell chambers to construct an in vitro VE barrier model.Cell voltage resistance meter was used to detect the transepithelial resistance(TER)in various groups after the bEnd.3 cells were infected with VSV at different time points;fluorescein isothiocyanate-dextran leakage assay was used to detect the permeability coefficients of the cells in various groups;immunofluorescence staining was used to observe the localization changes of VE-cadherin,β-catenin,and phosphorylated β-catenin(p-β-catenin)in cytoskeleton and adherens junctions(AJs)of the bEnd.3 cells after VSV infection;real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of Wnt and β-catenin mRNA in the cells in various groups;Western blotting method was used to detect the expression levels of Wnt,β-catenin,and p-β-catenin proteins in the cells in various groups.Results:The TCID50 of VSV was 10-4.5·100 μL-1.TheTranswell chamber experiment results showed that compared with infection 0 h group,the TERs in the cells in the other groups were significantly decreased(P＜0.05),and the permeability coefficients were significantly increased(P＜0.05).The immunofluorescence staining results showed that compared with control group,the cytoskeleton of the bEnd.3 cells in infection group was disordered,the cell gaps was increased,the linear index of AJs was significantly decreased(P＜0.05),and β-catenin and p-β-catenin translocated from the cell membrane to the perinuclear area.The RT-qPCR results showed that compared with infection 0 h group,the expression levels of Wnt mRNA in the cells in the other groups were significantly decreased(P＜0.05),while the expression levels of β-catenin mRNA showed no statistically significant difference(P＞0.05).The Western blotting results showed that compared with infection 0 h group,the expression levels of Wnt protein in the cells in the other groups were significantly decreased(P＜0.05),the expression levels of β-catenin showed no statistically significant differences(P＞0.05),and the expression levels of p-β-catenin were significantly increased(P＜0.05).After inhibiting the VSV replication and correcting the low density lipoprotein receptor(LDLR)abnormalities,the Transwell chamber experiment results showed that compared with infection group,the TER in the cells in correction group was significantly increased(P＜0.05),and the permeability coefficient was significantly decreased(P＜0.05).The immunofluorescence staining results showed that compared with infection group,the gaps in the cells in correction group were reduced,and the perinuclear aggregation of β-catenin and p-β-catenin in the cells was restrained.The RT-qPCR results showed that compared with infection group,the expression level of Wnt mRNA in the cells in correction group was significantly increased(P＜0.05).The Western blotting results showed that compared with infection group,the expression level of Wnt protein in the cells in correction group was significantly increased(P＜0.05),the expression level of β-catenin showed no statistically significant difference(P＞0.05),and the expression level of p-P-catenin was significantly decreased(P＜0.05).Conclusion:VSV infection can cause the LDLR inactivation,reduce the expression level of Wnt protein,increase the phosphorylation level of β-catenin and cause its internalization,disrupt the stability of AJs,and ultimately lead to VE barrier damage.

Objective:To investigate the strategy of balanoplasty in penile reconstruction with free forearm flap.Methods:From October 2010 to October 2015, 20 patients (19-41 years old) underwent penile reconstruction with free forearm skin flap in Xijing Hospital of Air Force Military Medical University, including 14 cases of transexuals (female to male), 3 cases of hermaphroditism, 2 cases of congenital penile dysplasia and 1 case of penile defect after trauma. According to the tissue quantity of forearm flap and the development of glans, three methods were adopted to reconstruct the glans: free forearm skin flap + E-shaped cartilage framework, free dorsalis pedis flap + E-shaped cartilage framework or free glans transplantation. The patients were followed up for a long time postoperation to observe the morphological changes of the glans after different method of balanoplasty.Results:Among the 20 patients, 18 cases were treated with distal part of free forearm flap and E-shaped cartilage framework for glans reconstruction, 1 case of transexuals used free dorsalis pedis flap and E-shaped cartilage framework for glans reconstruction, and 1 case of hermaphroditism used free glans transplantation to reconstruct glans. After 10-36 months of follow-up, all reconstructed penises survived. The shape of glans reconstructed by dorsalis pedis flap was the most plump and natural with clear coronal sulcus; the amount of glans tissue reconstructed by forearm skin flap was slightly insufficient, and the coronal sulcus was slightly flat, while the overall shape was satisfactory. The reconstructive glans with free glans transplantation was not plump, but they retained the shape of glans, which was lifelike.Conclusions:If the tissue quantity of forearm skin flap is sufficient, the distal part of free forearm flap and E-shaped cartilage framework is used; if the tissue quantity of forearm is seriously insufficient, the free dorsalis pedis flap and E-shaped cartilage framework is used. For hermaphroditism patients with well-developed glans, autologous glans free transplantation is used.Individualized strategy of balanoplasty according to different conditions of patients is conducive to obtaining satisfactory appearance of the glans.

Objective:To investigate the strategy of balanoplasty in penile reconstruction with free forearm flap.Methods:From October 2010 to October 2015, 20 patients (19-41 years old) underwent penile reconstruction with free forearm skin flap in Xijing Hospital of Air Force Military Medical University, including 14 cases of transexuals (female to male), 3 cases of hermaphroditism, 2 cases of congenital penile dysplasia and 1 case of penile defect after trauma. According to the tissue quantity of forearm flap and the development of glans, three methods were adopted to reconstruct the glans: free forearm skin flap + E-shaped cartilage framework, free dorsalis pedis flap + E-shaped cartilage framework or free glans transplantation. The patients were followed up for a long time postoperation to observe the morphological changes of the glans after different method of balanoplasty.Results:Among the 20 patients, 18 cases were treated with distal part of free forearm flap and E-shaped cartilage framework for glans reconstruction, 1 case of transexuals used free dorsalis pedis flap and E-shaped cartilage framework for glans reconstruction, and 1 case of hermaphroditism used free glans transplantation to reconstruct glans. After 10-36 months of follow-up, all reconstructed penises survived. The shape of glans reconstructed by dorsalis pedis flap was the most plump and natural with clear coronal sulcus; the amount of glans tissue reconstructed by forearm skin flap was slightly insufficient, and the coronal sulcus was slightly flat, while the overall shape was satisfactory. The reconstructive glans with free glans transplantation was not plump, but they retained the shape of glans, which was lifelike.Conclusions:If the tissue quantity of forearm skin flap is sufficient, the distal part of free forearm flap and E-shaped cartilage framework is used; if the tissue quantity of forearm is seriously insufficient, the free dorsalis pedis flap and E-shaped cartilage framework is used. For hermaphroditism patients with well-developed glans, autologous glans free transplantation is used.Individualized strategy of balanoplasty according to different conditions of patients is conducive to obtaining satisfactory appearance of the glans.

Objective: This study aimed to present our clinical experience of the utilization of remnant ear for the inferior part and lobule reconstruction in microtia patient with the fully pre-expansion ear reconstruction technique. Methods: During 2017.01—2018.12, 156 cases of unilateral ear reconstruction were performed. The operation was divided into three stages. Six months after the ear reconstruction, the third stage of the remnant ear utilization was performed. The surgery was planned according to the microtia classification and the remnant morphology. Results: All the patients had no complications. Follow-up period ranged from 7—24 months, and ten months on average. All the patients with typeⅠmicrotia had satisfying results of the inferior part and lobule reconstruction. Sixteen cases of types Ⅱand Ⅲ microtia patients had unfortunate outcomes, because the residual ears were too small to reconstruct the earlobe, and the incisure appeared when remnant and reconstructed ear was connected in cases of severe remnant malposition. The rest of patients were satisfied with the reconstruction. Conclusions The utilization of the remnant ear based on the types of microtia and the remnant morphology could achieve the satisfying result of reconstruction for most of the microtia patients.

Statins are collectively referred to as hydroxymethylglutaryl-CoA (HMG-CoA) reductase inhibitors.They exert powerful cholesterol-lowering effects by inhibiting key steps in the sterol biosynthetic pathway.At present,nearly 200 million people worldwide use statins.As clinical research progresses,the pleiotropic effects of statins are gradually discovered,especially those in the prevention and treatment of diseases of the circulatory system,nervous system and digestive system with high incidences in the elderly.We should to be alert to the adverse effects of statins and pay attention to the pleiotropic effects of statins in the elderly at the same time.

Objective To investigate the expression and effect of miR-22-3p in non-small cell lung cancer (NSCLC). Methods The miR-22-3p expression level in seventy-six NSCLC tissues and para-cancer tissues was detected by qRT-PCR. The relationship between the expression of miR-22-3p and gender,age,tumor size,histolo-gy grade,pathological type and lymph node metastasis was analyzed. The function of miR-22-3p on the prolifera-tion of NSCLC cells was tested by growth curve assay. Target genes of miR-22-3p were predicted by online software Targetscan. Luciferase reporter assay and qRT-PCR was used to certificate the prediction. Results The expression of miR-22-3p was increased in NSCLC tissues than the para-cancer tissues and was correlated to lymph node metas-tasis. Overexpression of miR-22-3p could suppress the proliferation of A549 cells. Astrocyte-Elevated Gene-1(AEG-1) was predicted to be a target of miR-22-3p. MiR-22-3p was revealed to bind to AEG-13′UTR by luciferase report-er assay. Overexpression of miR-22-3p could inhibit the expression of AEG-1 in A549 cells. Suppression of miR-22-3p could increase AEG-1 expression. Conclusion MiR-22-3p could inhibit the proliferation of NSCLC by tar-geting AEG-1.

Objective To study whether methicillin-resistant Staphylococcus aureus (MRSA)will increase the burden of patients with hospital-acquired pneumonia (HAP).Methods Patients with Staphylococcus aureus HAP in a hospital between January 1 ,2013 and November 31 ,2014 were selected,patients with MRSA HAP were as case group,patients with methicillin-sensitive Staphylococcus aureus (MSSA)HAP were as control group,propen-sity score matching (PSM)analysis were conducted to compare the prognosis of MRSA HAP and MSSA HAP (length of hospital stay, duration from infection to discharge, mortality, total therapeutic cost ). Results APACHE II score in case group was higher than control group before PSM was conducted,length of hos-pital stay and duration from infection to discharge were both longer than control group (40[20,94]d vs 28[21 ,53] d;19[10,46]d vs 17[8,29]d,respectively,both P <0.05).APACHE II score in case group and control group af-ter conducting PSM was not significantly different(P >0.05 ),data were balanced and comparable;there were no significant difference in length of hospital stay between two groups (28[21 ,52]d vs 28[21 ,53]d),duration from in-fection to discharge (15[9,25]d vs17[8,29]d),mortality(10.87% vs 15.22%),and total therapeutic cost (121 013.5[80 747.21 ,176 200]yuan vs 119 911 .2[66 994.08,241 184.7]yuan)(all P >0.05).Conclusion APACHE II score is an important factor affecting prognosis;after balancing this factor,there is no difference in the prognosis of patients with MRSA and MSSA HAP,MRSA HAP can not increase the burden of disease.

Objective To explore the significance of detecting miR-145 hypermethylation in non-small cell lung cancer (NSCLC). Methods Seventy-five NSCLC tissues and adjacent tissues was collected from May 2012 to January 2015. The methylation status of miR-145 promoter was tested by bisulfite sequencing PCR (BSP) and the expression of miR-145 evaluated by ISH. The correlation between miR-145 methylation and clinical parameters , and the relationship between miR-145 methylation and expression were statistically analyzed. Results The hypermethy-lation ratio of miR-145 in NSCLC tissues was higher than that in the adjacent tissues. Hypermethylation of miR-145 was correlated with NSCLC differentiation stage, but not with gender and age. In NSCLC, miR-145 hypermethyla-tion was negatively related with its expression. Conclusion MiR-145 hypermethylation is closely related with dif-ferentiation stage in NSCLC tissues. MiR-145 hypermethylation may be a potential biomarker for estimating NSCLC differentiation stage.

Objective To investigate the relationships among free triiodothyronine( FT3 ), free thyroxine (FT4 ), and thyroid-stimulating hormone( TSH) in both plasma and breast milk of patients with thyroid diseases. Methods A total of 102 female subjects with hyperthyroidism(GD), normal thyroid function(NC), and Hashimoto′s hypothyroidism(HT or hypothyroidism)were enrolled. Their plasma and breast milk were collected for measurement of FT3 and FT4 , and TSH. Meanwhile, 11 infants of patients with hyperthyroidism and another 11 infants of patients with hypothyroidism were selected, blood FT3 , FT4 , and TSH content were determined during lactating period and 2 months after lactation. Results (1) FT3 and FT4 contents in breast milk among 3 groups were different[(1. 48 ± 0. 81), (7. 79 ± 3. 56), and (0. 77 ± 0. 42)pg/ ml; (2. 94 ± 1. 43), (14. 78 ± 7. 40), and (1. 51 ± 0. 40)pg/ ml, P<0. 05], TSH in breast milk was similar between hyperthyroidism and normal groups(P>0. 05). (2) FT3 ratio of breast milk to plasma of the hyperthyroidism group was different to other 2 groups(0. 42 ± 0. 04 vs 0. 35 ± 0. 03, 0. 36 ± 0. 03, P<0. 05), but no difference existed in FT4 and TSH among 3 groups(both P>0. 05). (3)Blood FT3 , FT4 , and TSH contents from infants of patients with hyperthyroidism and hypothyroidism were different, both during lactating period and 2 months after lactation[(5. 06 ± 1. 76)vs (6. 51 ± 2. 23)pg/ ml, (17. 39 ± 2. 78)vs (19. 87 ± 3. 26)pg/ ml, (1. 34 ± 1. 33)vs (0. 74 ± 0. 78)mIU/ L; (1. 43 ± 0. 74)vs (1. 83 ± 0. 91)pg/ ml, (4. 28 ± 1. 55)vs (5. 00 ± 1. 75)pg/ ml, (6. 48 ± 2. 70) vs (5. 49 ± 2. 39) mIU/ L; all P<0. 05]. (4) FT3 and FT4 contents were positively correlated in plasma and breast milk(all P<0. 05), while TSH contents were positively correlated only in hypothyroidism group(P<0. 05). Conclusion FT3 , FT4 , and TSH in blood and breast milk are correlated.

Objective To analyze the present medical teaching textbooks and practice skill guidelines,and explore the profound causes of poor hand hygiene idea among doctors.Methods Three sets of unified textbook series used for domestic medical colleges and universities and two sets of manipulation skill guidelines were studies.Statistical method was conducted to analyze whether concepts and methods of hand hygiene,hand-washing and antiseptic han-drubbing were included in these teaching textbooks;as to eight aseptic manipulation skills,coverage of knowledge, steps of hand-washing and antiseptic handrubbing in manipulation skill guidelines were also analyzed.Results The mentioning rate of hand hygiene,hand-washing and antiseptic handrubbing in 8-year and 5-year program teaching textbooks were both 0 ,in nursing teaching textbooks was 1 00 % ;as to 8 aseptic manipulation in 2 sets of skill practice guidelines,mentioning rate of hand washing was 37 .50 % ,and method and steps of antiseptic handrubbing were both 0 .Conclusion School teaching and skill assessment are the basis,it is difficult to form the right idea by only relying on continuing education without basic education.Hand hygiene should be stressed in the written of teaching textbooks,guidelines should be written following the newest progress,so as to form the correct idea of hand hygiene among doctors.