Background The orbital fibroblasts (OFs) in thyroid associated ophthalmopathy (TAO) play important roles in the proliferative and inflammatory response.Seeking the drug which inhibit OFs growth is of a vital significance for the prevention and treatment of TAO.Research documented that colchicine has an anti-fibrosis effect.But its influence on OFs of TAO patient is few known.Objective This study was to investigate the effect of colchicine on growth and apoptosis of OFs in vitro.Methods The retroobital connective tissue was obtained form 3 TAO patients and cultured using explant method.OFs were passaged and identified by immunochemistry,and 3-8 genetaions of cells were used in the study.Colchicine at the concentrations of 1 × 10-8,1 × 10-7,1 × 10-6,1 × 10-5,1 ×10-4 mol/L was added into the RPMI 1640 with 10％ fetal bovine serum(FBS) to incubated the cells for 24,48 and 72 hours respectively,and only RPMI 1640 was used to culture the cells as the control group.Cell counting kit-8 (CCK-8)was used to detect the absorbance value (A450) of OFs for the evaluatuion of OFs and the inhibitory rate of colchicine to OFs.The colchicine of 1 ×10-6,1 ×10-5,1 × 10-4 mol/L was added into the culture medium for 48 hours,and then the apoptotic rate of the cells and the cell percentage in various cellular cycle was assayed by flow cytometry(FCM).The expression of transforming growth factor-β (TGF-β)in the cells was detected by immunochemistry to assess the influence of colchicine on the serection of the cells.Results Cultured cells showed the spindle-like in shape and the cell number was significantly increased with the incubation time.After incubated with 1 × 10-4,1 × 10 5,1 × 10-6,1 ×10-7,1 × 10-8 mol/L colchicines,the A450 values were gradually reduced with the increase of the concentrations of colchicine(F ion =62.004,P＜0.05),and significant differences were found between different contrations of colchicine groups(all P＜0.05).Aslo,gradually declined A450 values of the cells were seen with the lapse of culture time among the groups(Ftime =459.582,P＜0.05).The inbitory rate of colchicine to the cells was elevated with the increase of concentrations.The apoptotic rates of the cells were (1.73 ± 0.15) ％,(21.04 ± 4.56) ％,(31.84 ±6.21)％and(35.32±5.56)％ in the control group and 1 × 10-6,1 × 10-5,1 × 10 4 mol/L colchicine groups respectively,with statistically significant difference among the 4 groups (F =83.905,P＜0.05).With the increase of concentrations of colchicines,the cell percentage in G2 +M phase lessened gradually,showing significant difference among the control group and the 1 × 10-6,1 × 10-5,1 × 10-4 mol/L colchicine groups (F =20.443,P＜0.05).The expression of the TGF-β in the cells was (97.60± 2.09) ％ in the control group,and that in the 1 × 10-4 mol/L colchicine group was (44.43 ± 3.96) ％,presenting a significant difference between them (t =65.330,P ＜ 0.05).Conclusions Colchicine can induce apoptosis of OFs and inhibit the prolilferation of OFs in a time-and dose-dependent manner probably by decreasing the TGF-β secretion

Background Inflammation and adipogenesis are two parallel processes with increasing activity in severe thyroid-associated ophthalmopathy(TAO),and mevastatin was proved to have the inhibiting effect on the differentiation of adipose.Objective The aim of this work was to investigate the effects of mevastatin on the expression of cyclooxygenase-2(COX-2)and peroxisome proliferator activated receptor-γ(PPAR-γ)and differentiation of TAO-derived orbital preadipocytes,and explore its modulation effects on lipopolysaccharide(LPS)induced inflammation and the differentiation of TAO-derived orbital preadipocytes in vitro.Methods The retroorbital adipose tissue was obtained from 4 TAO patients during the surgery.The orbital fibroblasts were cultured from orbital adipose tissues using explant culture method.To study the suppressing effect of mevastatin on inflammatory response,cultured cells were divided into 5 groups.The 1000 μg/L LPS orbital fibroblasts were stimulated for 8 hours in group A,and 1000 μg/L LPS combined with 5 μmol/L,10 μmoL/L or 20 μmoL/L mevastatin were used respectively for the substitute in the group B,group C and group D.The orbital fibroblasts in group E were cultured routinely without any intervention as control.To observe the inhibiting effect of mevastatin on the differentiation of adipose,the group A were then subdivided into group A1-A6.After 1000 μg/L LPS was used to treat the cells for 8 hours,the ceils were induced to differentiate into adipocytes.All orbital preadipocytes from A1 to A6 were stimulated to differentiate into mature adipocytes with cocktail differentiation medium for a 10-day duration.During the procedure,group A2,A3 and A4 were interfered with 5,10 or 20 μmol/L mevastatin,and in the group A5 and A6,10 μmol/L mevastatin were added at the fourth day or eighth day.Intracellular fat accumulation in differentiated adipocytes was determined by oil red O staining.The absorption(A492 nm)was measured in the ceils by enzyme-linked immunosorbent assay(ELISA).Expression of COX-2 and PPAR-γ mRNA was detected by reverse transcription polymerase chain reaction(RT-PCR),and the expression of COX-2 and PPAR-γ protein was detected by Westernblot.The level of PGE2 in the supernatant was detected by ELISA.Results The expression of COX-2 protein and mRNA as well as the PGE2 levels in B,C,D group decreased markedly in comparison with those in A group(P＜0.05).With the increase of mevastatin concentration,the expression of COX-2 protein and mRNA as well as the PGE2 levels in B,C,D groups decreased successively(F =228.380,101.745,1586.881,P＜0.05).The expression of COX-2 protein and mRNA and PGE2 levels in E group were lower significantly than those in A,B and C groups(P＜0.05),but no significant differences were found between E group and D group(P＞0.05).The A492 value and the expressions of PPAR-γ protein and mRNA in differentiated cells showed the successively decrease in A1-A4 group with the elevation of mevastatin concentration(P＜0.05),and the evidently decreased A492 value and the expressions of PPAR-γ protein and mRNA also were seen in A1 and A5 groups compared with A3 group(P ＜ 0.05).Conclusions Mevastatin inhibits LPS-induced COX-2 expression,PPAR-γ expression,PGE2 secretion and differentiation of TAO-derived orbital fibroblasts in vitro in dose-dependent manner.Mevastatin plays these effect more prominently in early stage of adipocytes differentiation.

Objective To study the kidney histological categories of isolated hematuria in children.Methods Twenty-three children with isolated hematuria were performed renal biopsy under real time ultrasound guidance utilizing menghini style negative pressure biopsy device after local anesthesia or general anesthesia.The renopuncture tissue was directly sent by the mail-boxes to the remote pathologic service.All of the biopsies were examined by light microscopy,electron microscopy and immunohistochemistry.Results Biopsies were classified as measangial proliferative glomerulonephritis(MsPGN)(8 cases),minimal change nephropathy(MCN)(5 cases),IgA nephropathy(IgAN)(4 cases),thin basement membrane nephropathy(TMN)(3 cases),Alport′s syndrome(AS)(1 case),focal segmental glomerulosclerosis(FSGS)(1 case)and IgM nephropathy(IgMN)(1 case).Conclusions In this series,MsPGN,MGA,IgAN are the most common biopsy diagnosis.TMN and Alport′s are account for some proportion.A few IgMN and FSGS may also present as isolated hematuria.

Objective To study the effect of dexmedetomidine and midazolam on hemodynamics and sedation in patients with nasal intubation.Methods Forty patients whose ASA grade Ⅰ-Ⅱ and anticipated difficult airway were randomly divided into dexmedetomidine group(group D,20 cases)and midazolam group(group M,20 cases)according to the admission number.In group D,dexmedetomidine 1 μ g/kg were constant speed pumped in 10 minutes.In group M,midazolam 0.03 mg/kg were intravenous injected.Then nasal intubation were carried.Systolic blood pressure(SBP),diastolic blood pressure(DBP),mean arterial blood pressure(MAP),heart rate(HR),pulse oxygen saturation(SpO2),Ramsay sedation score,rate-pressure product(RPP),tip perfusion index(TPI)were recorded and compared before anesthesia (T0),fiberoptic bronchoscope pass by later nostril(T1),to spy on epiglottis(T2),intubation succeed(T3),after intubation 1 minute(T4)and after intubation 3 minutes(T5).Airway score and postoperative visit were evaluated.Results SBP,DBP,MAP,HR,RPP in group M were significantly higher at T1-T3 than those at T0 (P＜0.05),and were significantly higher than those in group D at the same time(P＜ 0.05).There was no significant difference in group D(P ＞ 0.05).Ramsay sedation score and TPI at T1-T3 in group M were significantly lower than those at To(P ＜0.05).Ramsay sedation score and TPI at T1-T5 in group D were significantly higher than those at T0(P ＜ 0.05),and were significantly higher than those in group M at the same time(P ＜ 0.05).The rate of airway score 1 score and intubation satisfaction in group D were significantly higher than those in group M[100％(20/20)vs.30％(6/20),90％(18/20)vs.50％(10/20)](P＜ 0.05).The rate of throat ache in group D was significantly lower than that in group M[5％(1/20)vs.35％(7/20)](P ＜0.05).Conclusions For difficult airway patients with nasal intubation during dexmedetomidine infusion,hemodynamics is stable and sedation is satisfied.

Objective To investigate the influence of the two-dimension computer-aided surgery navigation system to the concordance of lumbar spine pedicle screw fixation on both sides.Methods 1355 patients were undergone lumbar spinal pedicle screw fixation during January 2004 to December 2009.All patients were divided into tow groups:the navigation group (743cases) and the fluoroscopy assistant group (612 cases).All patients got standard A-P and lateral X-ray plate of lumbar spine within seven days after surgery.The X-ray images were analyzed by the software of Image-pro plus 5.0 to evaluate the concordance of lumbar spine pedicle screw fixation on both sides.The angle between axial line of pedicle screw and superior lamina terminals (α angle) and the angle between axial lines of pedicle screw on both sides (γangle) were measured.The position of the pedicle screw was checked weather it was in the lumbar pedicle partially.Results There were no significant differences (P＞0.05) between the α angle on both sides of L1-S5 vertebral body in navigation assistant group (L:3.89°±0.47°,R:3.94°±0.37°).The differences of the α angle on both sides of L2 (L:4.55°±1.27°,R:5.12°±1.87°) and L4 (L:4.22°±1.89°,R:6.62°±1.97°) vertebral body in the fluoroscopy assistant group had statistical significance (P＜0.05).There were no significant differences between the α angle on both sides of other bodies (L:4.32°±1.47°,R:4.37°±1.59°,P＞0.05).The γangle in navigation assistant group (2.32°± 0.27°) was obviously smaller than fluoroscopy assistant group (3.32°±1.51°),the differences had statistical significance (P＜0.05).Accuracy of pedicle screw in navigation assistant group was 91.5％ (3604/3938).Accuracy of pedicle screw in fluoroscopy assistant group was 87.6％ (2426/2768).The difference in accuracy of pedicle screw in both groups had statistical significance (x2=26.913,P＜0.0001).Conclusion The accuracy of pedicle screw and the concordance of pedicle screw on both sides can be significantly improved using the two-dimension perspective computer-aided surgery navigation system.

Objective To evaluate the value of 18F-FDG PET/CT in assessing radiosensitivity enhancement by irisquinone (IR) on rabbit xenografted VX2 lung tumor models.Methods Twenty-four tumor-beating rabbits were randomly divided into 3 groups (8 rabbits/group):group A with radiotherapy alone,group B with combined radiotherapy and IR,and group C without radiotherapy (the control group).18F-FDG PET/CT imaging was performed before radiotherapy and 24 h and one week after radiotherapy.The tumor SUVmax on delayed imaging was calculated in all rabbits.Two rabbits in each group were sacrificed after PET/CT imaging.HE staining was used to assess the differences in cancer cells among groups.Paired t test,one-way analysis of variance and Kaplan-Meier analysis were performed to analyze the data using SPSS 13.0.Results Before radiotherapy,the tumor SUVmax of all the 24 rabbits on standard and delayed imaging were 2.200 ± 0.761 and 3.162 ± 0.833 (t =-5.582,P ＜ 0.01).At 24 h post-radiotherapy,the delayed SUVmax of groups A,B and C were 2.614 ± 0.654,2.349 ± 0.869 and 5.663 ± 1.144,respectively.The differences between pre-radiotherapy and 24 h post-radiotherapy were statistically significant in all three groups (t =2.527,3.620,11.011,all P ＜0.05).One week after radiotherapy,the delayed SUVmax of groups A,B and C were 3.625 ± 1.064,3.058 ±0.850 and 7.424 ± 1.751,respectively.The differences among groups A,B and C were statistically significant (tA∶ B =2.652,tA∶C =3.799,tB∶C =4.366,all P ＜0.05).The cancer cells of group B were fewer than those of groups A and C by pathological findings,which was consistent with 18F-FDG PET/CT results.The survival times of groups A,B and C were (62.375 ±4.534),(69.000 ±4.660) and (54.125 ±5.276) d,respectively.Kaplan-Meier survival curves revealed better survival of group B as compared to groups A and C,respectively (Log-rank,x2 =7.355,16.943,both P ＜ 0.01).Conclusion 18 F-FDG PET/CT is able to evaluate the effect of irisquinone on tumor radiosensitivity enhancement.

Aim To observe the neuroprotective effect of different doses of propofol on ischemic fetal rat brain.Methods Eighteen healthy pregnant SD rats were randomly allocated into the following six groups with three rats in each.Group S: sham operation group, Group IR: ischemia/reperfusion group, Group P1~P3: different doses of propofol groups, Group B: bicuculline group.In group S and group IR, 1 ml saline solution was administered via caudal vein.In group P1~P3, 10, 30, 50 mg·kg-1 of propofol was administered via caudal vein respectively.In group B, when 50 mg·kg-1 propfol was administered via caudal vein, 5 mg·kg-1 bicuculline was injected intraperitoneally at the same time.Bilateral uterine ovarian arteries were clamped for 11 mins to make intrauterine distress model of the fetal rats.The brains of fetal rats were removed after 3 days of reperfusion.Brain sections(5 μm thick) were mounted and stained with Hematoxylin and eosin(HE).The profile of the hippocampus CA1 was evaluated under a light microscope and neuronal Lesion-index(LI) was calculated.MDA content of fetal rat brain was detected by thiobarbituric acid reaction method to determine the lipid peroxidation degree of brain.Results LI was (7.2±0.9) and MDA was (3.86±0.20) μmol·g-1 in group S.LI was 71.9±2.8 and the content of MDA was (9.10±0.45) μmol·g-1 in group IR, which increased significantly compared with those in group S(P<0.01).LI was (40.8±2.6), (21.4±1.4), (20.1±1.3) and the content of MDA was (7.32±0.41), (5.65±0.27), (5.44±0.28) μmol·g-1 in propofol groups, which decreased significantly compared with those in group IR(P<0.05).LI and the content of MDA was (51.2±2.3), (7.54±0.31) μmol·g-1 in group B,respectively, reversing partly the neuroprotevtive effect of propofl.Conclusion Propofol could protect the neurons in hippocampus CA1 region of fetal rat against intrauterine distress by reducing the concentration of MDA in the brain.

Objective:To explore the seizure recurrence and prognosis of epilepsy in relapse after anti-epileptic drugs (AEDs) withdrawal, and the influencing factors for these conditions.Methods:From December 2009 to August 2018, patients from the Affiliated Brain Hospital of Nanjing Medical University who relapsed after AEDs withdrawal were collected and followed up for at least 18 months. The seizure recurrence and prognosis of these patients were prospectively observed. The Kaplan-Meier method was used for survival analysis. The associated risk factors of the second relapse in the enrolled patients were analyzed by multivariate Cox analysis. The included patients were divided into good prognosis group and poor prognosis group according to whether they had achieved seizure freedom for at least one year after the first relapse. A multivariate Cox regression model was used to analyze the independent risk factors affecting their prognosis.Results:A total of 56 patients with epilepsy in relapse after AEDS withdrawal were collected. The average follow-up period was 46.23 months (18-120 months) from the initial time of seizure recurrence, and 21 patients (37.5%) had the second seizure recurrence. The relapsing risk in patients who continued to be observed without adding AEDs was higher than those who were treated immediately with drugs [9/16 vs 30.0% (12/40)], but without statistically significant difference (χ2=2.220, P=0.071). The results of univariate analysis showed that focal seizures, seizure frequency more than once per month before remission and poly-drug therapy before AEDs withdrawal were associated with high risk of the second relapse. Poly-drug therapy was an independent risk factor for the second relapse by multivariate Cox analysis ( HR=3.383, 95% CI 1.257-9.105). Of the 56 patients with epilepsy in relapse after AEDs withdrawal, 47 patients (83.9%) had a good prognosis without seizure for at least one year, and of 33 patients who were followed up for three years or more, 26 (78.8%) had no seizure for at least two years. Between the group retreated immediately after the first recurrence and the group without immediate treatment [87.5% (35/40) vs 12/16],there were no statistically significant differences on the proportions of good prognosis (χ2=2.333, P=0.258). Univariate analysis showed that the course of epilepsy>6 months before initial treatment, the frequency of seizures>1/month before remission, symptomatic epilepsy and poly-drug therapy were associated with the poor prognosis. However, none of independent risk factors was found for the poor prognosis through the multivariate analysis. Conclusions:The prognosis of patients with epilepsy in relapse after AEDs withdrawal is well, and about 2/3 patients with epilepsy in relapse after AEDs withdrawal have no more seizure recurrences. The poly-drug therapy before AEDs withdrawal may be an independent risk factor for the second seizure relapse.

Objective To explore the relationship between the mean platelet volume (MPV) and varicocele, so as to contribute to the etiopathogenesis of varicocele. Methods The MPV values were examined in 50 varicocele subjects and 46 controls with benign non-varicocele diseases. Parameters including MPV and their relation with varicocelewere analyzed in the two groups. Results The average ageof patients at the timeof examination was (23. 50 ± 5. 08) yearsold for varicocele subjects and (24. 67±4. 33) for controls (P = 0. 599). MPV value for varicocele subjectswas significantly higher than that for controls (P= 0. 030). MPV value was found to be significantly correlated with varicocele grade (r= 0. 497, P = 0. 03). Conclusion The MPV value is increased in varicocele patients and the varicocele grade is positively associated with MPV value.

Aim To study and compare the pharmaco-kinetic parameters of roxithromycin under normoxic and hypoxic rats. Methods A highly effective and rapid ultra-performance liquid chromatography with tandem mass spectrometry ( UPLC-MS/MS) method with posi-tive electrospray ionization source was successfully de-veloped and validated for quantification of roxithromy-cin in rat plasma. Sprague-Dawley rats were randomly divided into the hypoxia and normoxic groups. Each rat obtained a single dose of roxithromycin with 10 mg · kg-1 via intragastric administration. The pharmacoki-netic parameter comparison between normoxic and hy-poxic groups was calculated by SPSS software using in-dependent sample t test method. Results The main pharmacokinetic parameters of roxithromycin between the normoxic and hypoxic rats were:the AUC(0-t) 7 576 and 3 761 μg·h·L-1 , MRT(0-t) 5. 6 and 7. 7 h, T1/2 3. 4 h and 3. 9 h, CL 1. 5 and 3. 0 L · h-1 · kg-2 , tmax3. 1 and 3. 4 h, Cmax 1 116 and 372 μg·L-1 , re-spectively. The levels of Cmax and AUC of roxithromy-cin in hypoxic rats were statistically lower than those in normoxic rats. Conclusion The exposure level of rox-ithromycin in hypoxic rats markedly decreased. Our re-sults may provide an important experimental basis to adjust the dosage for roxithromycin in hypoxic clinical practice.