Objective To investigate effects of neural retina on development of the structure of outer blood retinal barrier in embryogenesis. Methods The retinal neural epithelium (RNE) and pigment epithelium (RPE) layers of 150, 120 and 90 embryonic chicken eyes incubated for 7, 10, and 14 days were peeled off. RNE was used to prepare the culture medium with different conditions (7drcSF3, 10drcSF3, 14drcSF3). RPE cells of 7- and 14-incubated chicken embryos were cultured on laminin-coated transwell filter. The SF3, 7drcSF3, 10drcSF3, 14drcSF3 medium were used respectively in the apical chamber and SF2 was used in basolateral chamber. After the formation of monolayer, the transepithelial electrical resistance of the RPE was detected. After the fixation of RPE cells, the condition of the tight junction among the cells was observed by immunohistochemistry and transmission electron microscopy. Results For the RPE cells of 7- and 14-day incubated embryonic eyes, the difference of TER in various medium of SF3/SF2, 7drcSF3/SF2, 10drcSF3/SF2, 14drcSF3/SF2 was statistically significant (P

Objective To compare the curative effect between minimal invasive internal fixation with Mast Quadrant and tra -ditional open internal fixation for treating thoracolumbar fractures .Methods A total of 46 cases suffered thoracolumbar fractures were randomly divided into the minimally invasive group (MQ) and the traditional open group (TO) ,the patients in MQ group re-ceived minimally invasive pedicle internal fixation under Mast Quadrant minimal invasive channel ;the patients in TO group received pedicle internal fixation under traditional open channel .Perioperative related indicators ,imaging indicators and improvements of low back pain were recorded and statistically compared respectively .Results The different of the volume of blood loss ,operation time and length of incision and postoperative volume of drainage between the two groups were statistically significant (P < 0 .05) ,the different of the volume of hospital duration ,postoperative VAS score between the two groups were statistically significant (P <0 .05) .The different of the volume of flange height in injured vertebral fanterior ,Cobb Angle between preoperative and postopera-tive were statistically significant (P< 0 .05) .And comparison between groups had no statistical significance (P> 0 .05) .Conclusion Compared with traditional open operation ,minimally invasive pedicle internal fixation under Mast Quadrant minimal invasive chan-nel has the advantage of more simple operation ,less intraoperative bleeding and postoperative pain less invasive ,fast recovery and short hospitalization stay .

OBJECTIVE:To determine the retative bioavailability of Diclofenac sodium eye-ointment in rabbit eyes and to evaluate its pharmacokinetics following topical application METHODS:48 rabbits were divided into two groups 0 1% Diclofenac sodium eye-ointment and Diclofenac sodium eyedrops were applied topically to rabbit eyes and aqueous humor and iridic tissue were collected at given times The concentration of Diclofenac sodium in tissue were measured by HPLC RESULTS:Diclofenac sodium eye-ointment group had higher concentration and longer half-time than Diclofenac sodium eyedrops group,however,the peak time were the same in two groups The relative bioavailabilities of Diclofenac sodium ointment in aqueous humor and iridic tissue were 183 33% and 205 68% respectively,compared with those of Diclofenac sodium eyedrops CONCLUSION:Diclofenac sodium eye-ointment can be absorbed well and released slowly,which can reduce the frequency of application of drug It is especially suitable for operation patient or application at night

Objective To evaluate the feasibility and validity of near infrared fluorescence imaging with indocyanine green in sentinel node biopsy for cN0 oral carcinoma.Methods Thirty cases of previously untreated oral carcinoma staged cT1-3 N0M0 were enrolled in this study.1 ml of indocyanine green (25 mg/ 5 ml) was injected both around the primary tumor in a 4 quadrant pattern and in the base of the tumor before skin incision.After elevation of the platysma flap and posterior retraction of the sternocleidomastoid muscle,fluorescence images were taken with a near infrared fluorescence detector until the hotspots were captured,then the hotspot lymph nodes were removed.Lymph nodes identified with fluorescent hotspots and verified in vivo were defined as sentinel nodes,and they were harvested and sent together with neck dissection specimen for pathologic study.Results Sentinel nodes were successfully harvested in all 30 cases.The number of sentinel nodes per case varied from 1 to 9,with an average number of 3.4.Routine pathology demonstrated that occult metastasis was exclusively found in the sentinel nodes in 5 cases (16.67％),and all the other lymph nodes were free from metastasis.No tracer associated adverse effects occurred in this series.Conclusion Near infrared fluorescence imaging with indocyanine green has a high detection rate in sentinel node biopsy for cN0 oral/oropharyngeal carcinoma and the sentinel nodes can evaluate the cervical metastatic status accurately.It is an easy,feasible and promising method,which is worthy of further investigation.

Objective:To analyze the risk factors of hepatocellular carcinoma microvascular invasion (MVI) and to construct a preoperative prediction clinical scoring system.Methods:A retrospective analysis was made on 113 patients with hepatocellular carcinoma undergoing hepatectomy at Zhongshan Hospital from March 2018 to Jun 2019.Postoperative pathology confirmed 35 cases with microvascular invasion.Results:The multivariate logistic regression model showed that the maximum tumor diameter( OR: 1.028, 95% CI: 1.001-1.005), the smoothness of the capsule edge( OR: 0.208, 95% CI: 0.062-0.699), the positive circulating tumor cells (CTC)( OR: 3.728, 95% CI: 1.029-13.501) and abnormal prothrombin(PIVKA-Ⅱ)( OR: 1.001, 95% CI: 1.000-1.002) were risk factors for MVI. The area, sensitivity and specificity of the clinical score constructed by assigning 1 point to each risk factor were 0.906, 74.29% and 92.31%, respectively. Clinical scores of 0, 1, 2, 3, and 4 predict MVI positive rates of 0 (0/26), 9.09% (3/33), 28.57% (6/21), 77.78% (14/ 18), 85.71% (12/14). Conclusions:Tumor maximum diameter>62 mm, PIVKA-Ⅱ>115 mAU/ml, unsmooth tumor capsule and CTC in peripheral blood are independent high risk factors in patients with MVI.

Objective:To investigate the correlation between preoperative circulating tumor cells (CTC) and microvascular invasion (MVI) in patients with hepatocellular carcinoma.Methods:The data of 227 patients who underwent hepatocellular carcinoma resection in Zhongshan Hospital Affiliated to Sun Yat-sen University from January 2018 to March 2020 were retrospectively analyzed. The peripheral blood CTC was detected by Cyttel detection before operation. The relationship between preoperative peripheral blood CTC and clinical characteristics of patients was analyzed; the multivariate logistic regression model was used to analyze the independent risk factors for MVI; the receiver operating characteristic (ROC) curve was used to compare the efficacy of each independent risk factor in predicting the occurrence of MVI, and the relationship between CTC and MVI was clarified.Results:According to the ROC curve, the cut-off values for predicting MVI of CTC, alpha-fetoprotein (AFP), protein induced by vitamin K absence or antagonist Ⅱ (PIVKA-Ⅱ), and tumor long-axis diameter were 3 CTC/3.2 ml, 158 μg/L, 178 AU/L and 59 mm. CTC-positive group had ≥3 CTC/3.2 ml in peripheral blood, and CTC-negative group had <3 CTC/3.2 ml, and there were 117 and 110 cases in the two groups. The median AFP levels of preoperative CTC-positive group and CTC-negative group were 123.0 μg/L (0-20 000.0 μg/L) and 9.6 μg/L (0-18 676.0 μg/L), and the median tumor long-axis diameter was 50.0 mm (5.0-200.0 mm) and 36.0 mm (2.0-150.0 mm), the differences between the two groups were statistically significant (both P < 0.05). Before operation, AFP≥158 μg/L ( OR = 3.551, 95% CI 1.426-8.843, P = 0.006), PIVKA-Ⅱ≥178 AU/L ( OR = 12.250, 95% CI 4.384-34.231, P < 0.01), peripheral blood CTC ≥ 3 CTC/3.2 ml ( OR = 8.913, 95% CI 3.561-22.306, P < 0.01) and tumor long-axis diameter ≥59 mm ( OR = 3.250, 95% CI 1.339-7.885, P = 0.009) were independent risk factors for the occurrence of MVI; the area under the ROC curve (AUC) of these factors for predicting MVI was 0.752, 0.777, 0.857 and 0.743. CTC was more effective in predicting MVI than AFP and tumor long-axis diameter, and the differences were statistically significant (both P < 0.05). The efficacy of CTC in predicting MVI was slightly better than that of PIVKA-Ⅱ, but the difference was not statistically significant ( P > 0.05). Conclusion:CTC may be one of the important indicators of hepatocellular carcinoma MVI in clinical practice.

Objective To obtain monoclonal antibodies ( mAbs ) against neutrophil gelatinase-associated lipocalin ( NGAL ) and a chemiluminescense immune quantification assay based one paired mAbs.Methods Six-to-eight weeks old female BALB/c mice were immunized with the purified recombinant human NGAL antigen( rhNGAL) that was produced by the Escherichia coil expression system.The spleen was fused with hybridoma for screening anti-NGAL monoclonal antibodies by indirect ELISA.Western blot was implemented to identify the reactivity with native NGAL. Results The rhNGAL antigen was found to form disulfide cross-linked dimers and present excellent immunogenicity.The reaction titer of the immune serum of NGAL immunized mice was about 106.Thirty mAbs were screened by indirect ELISA, hereinto;the EC50 values of mAb23C12 and 38D10 were 0.034 g/mL, 0.022 g/mL respectively.The antibodies pair, 38D10/23C12-SAE labeled with AcridiniumEster(AE), were shown to work well in chemiluminescense immune response quantitative detection which was screened by NGAL standardand clinical urine samples.This detection can resolve positive and negative samples with a statistically significant difference (P<0.0001).And the correlation coefficient R2between NGAL quantitative results and that of the Abbott's NGAL chemiluminescence immune assay kit was greater than 0.97.The detection linear range was 10-1500 ng/mL, analytical sensitivity of the method was 0.63 ng/mL.Conclusion Highly purified rhNGAL antigen and specific anti-NGAL monoclonal antibodies are generated in this study.The detection capability of method is comparable with that of the international commercial kit.

Objective: To investigate the impact of minimal residual disease (MRD) by multiparameter flow cytometry (MPFC) during aplasia on efficacy and prognosis of de novo acute myeloid leukemia (AML) (non M₃) patients. Methods: The MRD data by 8-color MPFC during aplasia (day 14-15 of induction therapy) in 85 de novo AML (non M₃) patients and the MRD impact on efficacy and prognosis were retrospectively analyzed. Results: Data of 85 patients, including 42 males (49.4%) and 43 females (50.6%) , were collected, with a median age of 35 (15-54) years. The median MRD by MPFC during aplasia was 0.58% (0-81.11%) , and 70 (82.4%) patients achieved complete remission (CR) after first induction chemotherapy. The cutoff of MRD by receiver operating characteristic (ROC) analysis was 2.305% (Se= 0.867, Sp=0.800) . The CR rate after one course was significantly higher in patients with MRD<2.305% [96.6% (56/58) ]than in patients with MRD≥2.305%[51.9% (14/27) ] (χ²=22.348, P<0.001) ; no significant difference with respect to relapse-free survival rate (χ²=1.08, P=0.299) or overall survival rate (χ²=0.42, P=0.516) could be demonstrated for the comparison of the two groups. Multivariates analysis showed MRD divided by 2.305% was the only independent prognostic factor for CR after one course (OR= 21.560, 95% CI 4.129-112.579, P<0.001) . Conclusion Flow cytometric MRD divided by 2.305% during aplasia could be a predictor of efficacy after first induction therapy in AML patients.

Objective:To This study was to investigate the expression and possible clinical significance of microRNA-146b (miR-146b) and signal transducer and transcriptional activator 1 and 3 (STAT1/3) in peripheral blood mononuclear cells (PBMCs) of patients with primary gouty arthritis (GA).Methods:The peripheral blood samples, clinical data and laboratory indexes of 120 male cases of GA [including 57 cases of acute (AG group) and 63 cases of intermittent (IG group)]and 66 healthy subjects (HC group) were collected. The expression levels of miR-146b and STAT1/3 in PBMCs were detected by real-time fluorescence quantitative PCR (RT-qPCR). The differences among the three groups were compared and the correlation between them and clinical indexes was analyzed. The receiver operating characteristic curve (ROC) was constructed to evaluate its diagnostic value in GA. After the PBMCs of 18 healthy subjects were stimulated by 100 μg/ml MSU for 3 hours to simulate acute gout inflammatory environment, the transcriptional changes of IL-1β, miR-146b and STAT1/3 were detected by RT-qPCR, and the expressions of IL-1β, STAT1/3 protein and phosphorylated protein were detected by Western blotting. T test or one-way ANOVA and LSD- t test were used in accordance with the normal measurement data, Kruskal-Wallis H and Mann-Whitney U test were used in the non-normal data, Spearman correlation analysis was used in the correlation between variables, and the diagnostic value was evaluated by the receiver working characteristic curve ROC. Results:①There were statistical differences in the expression of miR-146b, STAT1 and STAT3 among the three groups ( F=7.02、19.52、17.07, all P<0.001). The expression of miR-146b in gout group [(0.32±0.28)] was significantly higher than that in HC group (0.19±0.18)( t=2.96, P=0.003), while STAT1(0.019±0.012) and STAT3(0.014±0.010) were significantly lower than those in HC group (0.038±0.029),(0.025±0.016)( t=6.26, 5.56, both P<0.001). Further subgroup analysis showed that the expression of miR-146b in AG and IG groups was higher than that in HC group[(0.27±0.17), (0.38±0.35), (0.19±0.18), t=2.09, 3.30, both P<0.05], but that in AG group was lower than that in IG group ( t=2.02, P<0.05). The expression of STAT1 mRNA in AG and IG groups was lower than that in HC group [(0.020±0.012), (0.019±0.012), (0.038±0.029), t=4.89, 4.56, both P<0.001], but there was no statistical significance between AG and IG groups ( t=0.24, P>0.05). The expression of STAT3 mRNA in AG and IG groups was lower than that in HC group [(0.016±0.012), (0.012±0.008), (0.025±0.016), t=5.64, 3.33, both P<0.01], and the expression of STAT3 mRNA in AG group was higher than that in IG group ( t=2.12, P<0.05). ② Spearman correlation analysis showed that the expression of miR-146b in GA was negatively correlated with HCY ( r=-0.37, P=0.014), STAT1 was negatively correlated with Crea ( r=-0.29, P=0.019), positively correlated with eGFR ( r=0.25, P=0.047), and STAT3 was negatively correlated with HDL-C ( r=-0.27, P=0.033). ③ROC curve showed that the AUC (95% CI) of miR-146b, STAT1 and STAT3 were 0.679(0.582, 0.776), 0.710(0.629, 0.791) and 0.705(0.626, 0.783), and the combined AUC(95% CI) of the three was 0.836 (0.765, 0.907). ④Compared with blank control group and negative control group, the expression of miR-146b in PBMCs of 18 cases of HC was significantly decreased ( H=14.44, P=0.003), while the expression of IL-1β, STAT1 and STAT3 mRNA was significantly increased after 3 h of MSU stimulation ( H=26.44、27.26、15.90, all P<0.001). The expression of IL-1β, STAT1 and STAT3 protein and phosphorylated protein in the model group were significantly higher than those in the blank control group, and the differences were statistically significant ( t=9.97、6.63、7.48、11.25、6.28, all P<0.01). Conclusion:The abnormal expression of miR-146b and STAT1/3 in GA is related to some clinical indicators, suggesting that it may be involved in the regulation of gout immune inflammatory response and metabolism, and the specific mechanism is worth further study.