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To study the inhibitory effect of Nogo-A shRNA on cell line PC12, the Nogo-A shRNA (short hairpin RNA, or shRNA) was designed and synthesized. The annealed shRNA template was inserted into plasmid pGenesil-1 containing enhanced green fluorescent protein (EGFP) gene by gene cloning technique to generate eukaryotic expression vector. The recombinant plasmid was transfected into PC12 cells by lipofecamine2000 and the mRNA and protein expression level of Nogo-A gene was detected by RT-PCR and Western blotting 48 h after the transfection. Gene sequencing showed that that the Nogo-A shRNA eukaryotic expression vector was successfully constructed. No significant change was found in the Nogo-A mRNA and protein expression level in empty vector-transfected group as compared with controls (P>0.05), while the expression level in shRNA-transfected group decreased significantly (P<0.05). It is concluded that the pGenesil-1/Nogo-AshRNA recombinant plasmid can effectively suppress the expression of Nogo-A gene in PC12 cells.
Cloning, Molecular ; Gene Knockdown Techniques/*methods ; Genetic Vectors ; Green Fluorescent Proteins/genetics ; Myelin Proteins/*genetics ; Myelin Proteins/metabolism ; PC12 Cells ; Plasmids ; RNA Interference ; RNA, Messenger/genetics ; RNA, Messenger/metabolism ; RNA, Small Interfering ; Transfection

This study investigated the effect of TNP-470 in combination with carmustine (BCNU) on the growth of subcutaneously implanted human glioblastoma xenografts in nude mice. Human glioblastoma U-251 cells (1×10(7)) were injected into 24 nude mice subcutaneously. The tumor-bearing mice were randomly divided into 4 groups on the seventh day following tumor implantation: TNP-470 group, in which TNP-470 was given 30 mg/kg subcutaneously every other day 7 times; BCNU group, in which 20 mg/kg BCNU were injected into peritoneal cavity per 4 days 3 times; TNP-470 plus BCNU group, in which TNP-470 and BCNU were coadministered in the same manner as in the TNP-470 group and the BCNU group; control group, in which the mice were given 0.2 mL of the mixture including 3% ethanol, 5% acacia and 0.9% saline subcutaneously every other day 7 times. The tumor size and weights were measured. The tumor microvessel density (MVD) was determined by immunostaining by using goat-anti-mouse polyclonal antibody CD105. The results showed that on the 21th day following treatment, the volume of xenografts in the TNP-470 plus BCNU group was (108.93±17.63)mm(3), markedly lower than that in the TNP-470 group [(576.10±114.29)mm(3)] and the BCNU group [(473.01±48.04)mm(3)] (both P<0.01). And the xenograft volume in these 3 treatment groups was even much lower than that in the control group [(1512.61±470.25) mm(3)] (all P<0.01). There was no significant difference in the volume of xenografts between the TNP-470 group and the BCNU group (P>0.05). The inhibition rate of the tumor growth in the TNP-470 plus BCNU group was (92.80±11.37)%, notably higher than that in the TNP-470 group [(61.91±6.29)%] and the BCNU group [(68.73±9.65)%] (both P<0.01) on the 21th day following treatment. There was no significant difference in the inhibition rate of tumor growth between the TNP-470 group and the BCNU group (P>0.05). The MVD of xenografts in the TNP-470 plus BCNU group was decreased significantly as compared with that in the TNP-470 group or the BCNU group (both P<0.05). The MVD of xenografts in the 3 treatment groups was markedly reduced as compared with that in the control group (all P<0.05). No significant changes in weights were observed before and after the treatment in each group (all P>0.05). It was concluded that the combination of TNP-470 and BCNU can significantly inhibit the growth of human glioblastoma xenografts in nude mice without evident side effects.

Background The diseases of neuro-ophthalmology are common,which can influence the quality of patients' life seriously.The 39-item National Eye Institute Visual Function Questionnaire (NEI VFQ-39) is an instrument to assess self-reported visual impairment in studies of vision.However,until now,no studies have been performed to specifically assess the vision-related quality of life in persons with diseases of neuro-ophthalmology.Objective To evaluate vision-related quality of life for neuro-ophthalmology inpatients through the NEI VFQ-39,for the better treatment and nursing for them.Methods Ninety-seven neuro-ophthalmology inpatients were scheduled in research from June 1 to September 30,2012,and we collected all the clinical datas and the Chinese version NEI VFQ-39.The NEI VFQ-39 subscale item scores were compared among subgroups divided by monocular or binocular incidence,diseases and best corrected visual acuity(BCVA),and correlation analysis of the NEI VFQ-39 scores and BCVA for better-seeing and worse-seeing eyes was performed.Results The mean age of scheduled patients was (36.6±14.4)years,with no difference in gender.The mean composite score of VFQ-39 was 57.36(46.50,73.38),mental health subscale score was lowest [45.00 (35.00,60.00)].In the four eye diseases groups,except for eye pains,degree of dependence,periphery vision,anterior ischemic optic neuropathy (AION) group had lower scores compared with the other three groups,but there were no statistical significances among the four groups (all at P＞0.05).Except for eye pains,patients with monocular incidence had better scores than those with binocular incidence,with statistical significant significances between them(all at P＜0.05).Patients with BCVA in the better-seeing eye ＞ 20/32 had better scores than the other two groups,compared with the BCVA in the better-seeing eye ＜20/200 group,except for overall health and eye pains,cornpared with the 20/200 ≤ BCVA in the better-seeing eye ＜20/23 group,except for eye pains,social activity and color vision,and the rest subscale scores had significant differences among the three groups(all at P＜0.05).NEI VFQ-39 scores had positive correlations with the BCVA for better-seeing and worse-seeing eyes,and had strong correlations with the BCVA for better-seeing.Conclusions Neuro-ophthalmology diseases have serious influences on patients' visual function and quality of life.The quality of life has direct correlation with BCVA,so improving their visual function is helpful to their quality of life.

Objective To study the changes of gonadotropin releasing hormone agonist (GnRH-a) in pinopodes during luteal phase and to explore the possible mechanism of GnRH-a in luteal phase support of assisted reproductive technology (ART).Methods Totally 40 primary infertility women who were treated with ART due to male factors were enrolled,according to the order of the group they were randomly divided into experimental group and control group.On the 7th day after ovulation,the experimental group received a subcutaneous injection of 0.1 mg of GnRH-a,while the control group received a subcutaneous injection of placebo only (0.9％ salinc 2 ml),3 days later they came to the clinic again.Serum estradiol and progesterone levels were measured before and after treatment in each group.Pinopodes were collected for electron microscopic examination.Levels of ER and PR were detected by western blot.Results (1) There was no significant difference between the experimental group and the control group in the estrogen level before and after the treatment (all P＞0.05).The level of progesterone in the experimental group after treatment [(66.8± 14.9) nmol/L] was significantly higher than that before treatment(P＜0.05);also significantly higher than the same period of the control group (P＜0.05).(2) There was no significant difference in the expression of ER protein in the experimental group before and after treatment (P＞0.05).The expression of PR in the experimental group after treatment was significantly lower than that before treatment (P＜0.05);also lower than the same period of the control group (P＜0.05).(3) Expression amount of pinopodes in the experimental group after treatment was significantly higher than that before treatment [65％ (13/20) versus 25％ (5/20),P＜ 0.05],and the development trend was more mature [the percentage of maturation:75％ (15/20) versus 35％ (7/ 20),P＜0.05].Expression amount of pinopodes after treatment and the percentage of maturation in the experimental group were significantly higher than those in the same period of control group (P＜0.05).Conclusion GnRH-a in luteal phase support may play a role through the corpus luteum,which may promote the secretion of progesterone,downregulation of PR expression,promote the growth of pinopodes,and improve the endometrial receptivity.

Objective To investigate the role of Fas antigen,Fas ligand(FasL)and soluble Fas(sFas)in the pathogenesis of vitiligo.Methods The expression of Fas and FasL on peripheral blood lym-phocyte(PBLC)was detected by flow cytometry.Serum sFas was quantitated using a dual antibody sandwich enzyme-linked immunosorbant assay(ELISA).Results The expression of Fas and FasL was significantly de-creased in patients with vitiligo vulgaris(43.45%and58.40%)than those in normal controls(58.30%and64.65%)(P

Objective To analyze the value of the potential risk factors on predicting primary graft dysfunction (PGD) after bilateral lung transplantation for the patients with idiopathic pulmonary fibrosis (IPF).Methods A retrospective study was conducted. Fifty-eight patients with IPF who underwent the bilateral lung transplantation admitted to Wuxi People's Hospital Affiliated to Nanjing Medical University from June 2014 to March 2017 were enrolled. The grade 3 PGD happened within 72 hours after transplantation was taken as the outcome event, and these patients were divided into PGD and non-PGD groups. The age, gender, body mass index (BMI), underlying disease, and N-terminal-probrain natriuretic peptide (NT-proBNP) before operation, pulmonary artery systolic pressure (PASP), pulmonary artery diastolic pressure (PADP), and mean pulmonary artery pressure (mPAP) before and after operation, duration of operation, the volume of blood transfusion during operation and postoperation, the use of extracorporeal membrane oxygenation (ECMO) during the operation, blood purification treatment after operation, and shock within 3 days after operation were recorded. The differences of parameters mentioned above between the two groups were compared. The predictive factors of PGD were searched by binary logistic regression analysis, and the receiver operating characteristic curve (ROC) was plotted to analyze the predictive value of preoperative PADP for grade 3 PGD after transplantation.Results Among 58 patients who underwent the bilateral lung transplantation, 52 patients were enrolled. The rest patients were excluded because of incomplete clinical data. There were 17 patients in the PGDgroup, with a mortality rate of 47.06%. The non-PGD group included 35 patients with a mortality rate of 8.57%. PADP and mPAP ahead of operation, the dosage of red cells suspension after the operation, and the total amount of blood transfusion during and after the operation in PGD group were significantly higher than those in non-PGD group [PADP ahead of operation (mmHg, 1 mmHg = 0.133 kPa): 33.7±10.5 vs. 25.3±10.1, mPAP ahead of operation (mmHg): 40.4±14.1 vs. 32.8±11.1, the dosage of red cells suspension after the operation (mL): 700 (300, 1500) vs. 300 (300, 500), the total amount of blood transfusion during and after the operation (mL): 2250 (1850, 4275) vs. 1800 (1550, 2800)], with statistically significant differences (all P < 0.05). There were no significant differences in age, gender, BMI, underlying disease, NT-proBNP before operation, PASP before and after operation, PADP and mPAP after operation, duration of operation, amount of plasma and red cells suspension as well as total amount of blood transfusion during operation, plasma amount and total amount of blood transfusion after operation, amount of plasma and red cells suspension during and after operation, use of ECMO during operation, blood purification treatment after operation, and shock after operation between the two groups (all P > 0.05). It was shown by binary logistic regression analysis that the preoperative PADP was the independent risk factor of grade 3 PGD after lung transplantation [odds ratio (OR) = 1.084, 95% confidence interval (95%CI) = 1.016-1.156,P = 0.015]. It was shown by ROC curve that the area under the ROC curve (AUC) of the PADP before operation for predicting the grade 3 PGD after lung transplantation was 0.728. When the cut-off value was 36 mmHg, the sensitivity was 47.1%, and the specificity was 91.4%.Conclusions Compared with the non-PGD group, the patients with higher preoperative PADP were more common in the PGD group, and the patients in the PGD group were more likely to be characterized by grade 3 PGD after lung transplantation. The preoperative PADP was an effective predictor of grade 3 PGD after lung transplantation.

Objective To observe the influence of phentolamine on N terminal B-type natriuretic peptide precursor(NT-proBNP),blood gas analysis,hypersensitive c-reactive protein and plasma D-dimer in patients with chronic cor pulmonale Methods One hundred and one cases patients with chronic cor pulmonale were randomly divided into two groups,51 patients in the treatment group,50 patients in the control group.All patients were treated with normal anti-infection,eliminating phlegm to smooth wheezing antithrombotic for one week,as the same time patients in the treatment group were treated with phentolamine for one week.Through observed the treatment effect of phentolamine to chronic cor pulmonale,the level of NT-proBNP,blood gas analysis,hypersensitive c-reactive protein and plasma D-dimer in patients with chronic cor pulmonale before and after the treatment were analyzed.Results Compared with before treatment,the levels of NT-proBNP,PCO2,hypersensitive c-reactive protein,plasma D dimer were lower than after one week in two groups,while the level of PO2 was higher.Treatment group:NT-proBNP (1 712.76±572.32) ng/L vs.(271.59±163.05) ng/L,t=20.42,P<0.05,PCO2 (66.34±5.81) mmHg vs.(52.58±5.82) mmHg,t=16.46,P<0.05,PO2 (59.28±6.13) mmHg vs.(73.64±6.10) mmHg,t=23.02,P<0.05,hypersensitive c-reactive protein 86.0(28.0) mg/L vs.23.0(12.0) mg/L,Z=-6.22,P<0.05 mg/L,plasma D-dimer (4 953.37±1 654.09) μg/L vs.(1 847.90±838.66) μg/L,t=17.11,P<0.05.Control group:NT-proBNP (1 527.24±658.70) ng/L vs.(612.58±357.59) ng/L,t=14.52,P<0.05,PCO2 (65.41±5.23) mmHg vs.(56.46±5.65) mmHg,t=13.04,P<0.05,PO2(60.57±5.84) mmHg vs.(67.21±5.19) mmHg,t=-10.06,P<0.05,hypersensitive c-reactive protein 79.0(29.0) mg/L vs.43.0(20.0) mg/L,Z=-6.16,,P<0.05,plasma D-dimer (4 408.02±1 682.83) μg/L vs.(2 598.28±1 242.73) μg/L,t=12.15,P<0.05.But the levels of NT-proBNP,PCO2,hypersensitive c-reactive protein and plasma D-dimer reduced significantly,the level of PO2 increased more significantly in treatment group(t(z)=-6.19,-3.39,-7.16,-3.56,5.70,all P<0.05).Conclusion Phentolamine can reduce the level of NT-proBNP,PCO2,hypersensitive c-reactive protein and plasma D-dimer and increased the level of PO2 in patients with chronic cor pulmonale.Phentolamine combined with routine treatment can improve the clinical efficacy of patients with chronic cor pulmonale.

To establish diagnostic criteria for common traditional Chinese medicine (TCM) syndromes in osteoporosis.

To establish a quantitative estimate model for diagnosing traditional Chinese medicine (TCM) syndromes of patients with osteoporosis.