Objective To explore the effect of Paclitaxel-Chitosan Sustain film on growth, apoptosis and cell cycle of biliary fibroblasts cells. Methods Human biliary fibroblasts cells were cultured and treated with PTX-CSF and naked PTX,separately, untreated cells as blank control. The experiment was divided into five groups:untreated group, simple PTX-treated group (250nM) and low, medium and high chitosan sustained-release film PTX-treated group (100 nM, 250 nM, 500 nM). The proliferations of cells were determined by MTT assay. The apoptosis and cell cycle of cells were detected by FCM. Results The proliferation of biliary fibroblasts cells was inhibited by PTX-CSF with time-dependent and dose-dependent, and the inhibiting effect was more obvious than naked PTX treatment as the time went on. Meanwhile, PTX-CSF could inhibit the magration of bile duct fibroblasts induced by TGF-β1,and had longer effect than naked PTX. After 72 h, the apoptosis rate of cells treated with PTX-CSF was significantly higher than cells treated with naked PTX or untreated cells(P<0.05), the difference between naked PTX or untreated cells was not significant. Compared with untreated cells, the proportion of G 2/M in cells treated with PTX or PTX-CSF were significantly increased, and the former was sinificantly higher than the latter(P<0.05). Conclusion Compared with naked PTX, PTX-CSF has strong cytotoxic effects and obviously sustained-release effect. The effective concentration can be maintain for a long time by PTX-CSF, and it could be as the novel drug delivery system to continuously inhibit proliferation of bile duct fibroblasts.

Objective To discuss the best resistance factors of paclitaxel(Taxinol)on benign biliary scar fibrosis,in order to provide an effective basis for clinical prevention and treatment of benign biliary scar fibrosis.Methods Human bile duct epithelial cells were cultured in vitro,the prepared PTX at 0.001 uM,0.005 uM,0.1 uM,0.5 uMand 1 uMconcentration were separately added into cells for 48 h.The half inhibitory rate of BEC (IC50) were determined by MTT and the optimal concentration were confirmed.Human bile duct epithelial cells were cultured in 0 h,24 h,48 h and 72 h,the inhibitory rate of BEC at 100 nM,250 nM,and 500 nM PTX-Chitosan Sustained release membranes and the optimal concentration of PTX were determined by MTT and the optimal concentration of PTX-SRM were obtained.Human bile duct epithelial cells were cultured for 48 h and 72 h,the mRNA and protein expression ofα-SMA,E-cadherin,Vimentin were detected by Western Blot and Real-time PCR methods.Results The optimum resistance concentration of PTX to benign biliary scar was 250 nM.PTX and PTX-SRM could effectively inhibit the proliferation and transformation of BEC,and the best effective treatments to resist benign biliary scar fibrosis were low and middle concentrations of PTX-SRM,the best drug loading were 100 nMand 250 nM.The inhibition duration of PTX-SRMon BEC was longer than PTX alone(P<0.05).Conclusion The inhibition of PTX-SRMon BEC proliferation and transformation is better than the single drug of PTX,which provides a new scientific and feasible method for clinical prevention and treatment of benign biliary scar.

Objective:To introduce the experience and clinical efficacy of a new type of negative pressure suction ureteroscope in the treatment of upper ureteral calculi, and compare it with traditional rigid ureteroscope and flexible ureteroscope lithotripsy.Methods:From January 2018 to January 2020, 160 patients with upper ureteral incarcerated calculi who were treated in Shengjing Hospital Affiliated to China Medical University were included in this study, and their clinical data were retrospectively analyzed.There were 50 cases in the rigid ureteroscope group, 54 cases in the flexible ureteroscope group and 56 cases in the new negative pressure combined ureteroscope group.The differences of operation time, hospital stay, stone free rate and postoperative complications were observed and compared among the three groups.According to whether the maximum diameter of the stone is more than 1.5 cm, the subgroups were analyzed to further compare the curative effect of the three groups.Results:All the 160 operations were completed successfully and none of them were converted to open surgery.The stone free rate of the new negative pressure suction ureteroscope Group (90.0% (18/20))was higher than that of the flexible ureteroscope Group (61.9% (13/21)) and the rigid ureteroscope Group (55.6% (10/18)), χ 2=8.49, P<0.001). The stone free rate in the first month after operation was also superior (96.4% (54/56), 77.7% (42/54), 74.0% (37/50), χ 2=5.48, P=0.01). The stone subgroup analysis showed that when the stone diameter increased (>1.5 cm), the net stone rate of the new negative pressure suction ureteroscope Group [91.6% (22 / 24)] was better than that of the flexible ureteroscope Group (62.5% (15/24)) and the rigid ureteroscope Group (50.0 (8/16))(χ 2=6.480, P=0.001). Conclusion:Compared with the traditional ureteroscopic lithotripsy, the new negative pressure suction ureteroscopic lithotripsy has higher initial and overall stone free rate, and higher safety factor.For the patients with stone diameter>1.5 cm, the new negative pressure suction ureteroscope lithotripsy has more advantages.

Objective To investigate the influence of abaI expression on acinetobacter baumannii biofilm formation.Methods Acinetobacter baumannii strain S isolated from bums patients was collected for the study,while the standard strain ATCC19606 was served as the control.At 6,24 and 48 hours,the gene expressions of abaI,pgaA,pgaB and pgaC were detected by real-time fluorescent quantitative PT-PCR,secretion of N-acyl-homoserine lactones (AHLs) by biological sensor and biofilm formation by MTT method.Results (1) Gene expressions of abaI,pgaA,pgaB and pgaC at 6 hours were 8.63 ±5.93,1.98 ± 1.93,1.01 ± 1.32 and 2.67 ± 3.46 respectively,which showed a quick increase at 24 hours (22.81 ± 17.60,5.13 ± 4.32,5.66 ± 3.97,11.97 ± 7.75 respectively),followed by a rapid decline in 48 hours (3.43 ± 0.88,1.30 ± 0.24,3.01 ± 3.00,3.02 ± 3.29 respectively).Gene expressions of pgaB and pgaC at 6 hours and that of pgaA and pgaC at 48 hours revealed statistically significant differences from those at 24 hours (P ＜ 0.05).(2) AHLs showed a level of 18.49 ± 11.03 at 6 hours,reached a peak of 52.23 ± 15.95 at 24 hours,then descended to 5.53 ± 0.94 at 48 hours.AHLs level at 24 hours showed statistically significant difference from that at 6 hours and 48 hours (P ＜ 0.05).(3)Biofilm formation at 24 hours and 48 hours was 2.83 ±0.44 and 2.71 ±0.15 respectively,far higher than that at 6 hours (0.49 ± 0.11,P ＜ 0.05).(4) In the correlation analysis among AHLs,biofilm formation and gene abaI,pgaA,pgaB and pgaC expressions,significant positive correlation was found between abaI and pgaA and between AHLs and pgaC expression (P ＜ 0.05).Conclusion Acinetobacter baumannii may regulate gene expressions of pgaA and pgaC responsible for biofilm formation to adjust to the external environment by means of changing abaI gene expression and AHLs secretion.

AIM: To explored the treatment effects of retinal laser photocoagulation and the applications of color Doppler flow imaging(CDFI) in the diabetic retinopathy.METHODS: We collected 60 patients(120 eyes) with diabetic retinopathy(Ⅲ~Ⅳstage) from February 2013 to February 2014 in Anyang Eye Hospital admitted in fundus disease department.The health control (HC) group of 55 normal people was established simultaneously.Ocular blood flow velocity of the 55 normal people (110 eyes) and the 60 patients (120 eyes) was examined by CDFI of central retinal artery (CRA) and posterior ciliary artery (PCAs) using the PHILIPS HD6.Peak systolic velocity(PSV) was recorded.The examinations of CDFI, electroretinogram(ERG) and the vision were performed in pre-operation, 1,7d, 1, 3 and 6mo postoperatively.RESULTS: Compared the PSV of CRA of the DR group before and after photocoagulation to the data of the HC group, there was a significant difference(P<0.05);compared the PSV of CRA of the DR group after photocoagulation with the data before, there was a significant difference(P<0.05).Compared the PSV of PCAs of the DR group before and after photocoagulation to the data of the HC group, there was a significant difference(P<0.05);compared the PSV of PCAs of the DR group after photocoagulation with the data before, there was a significant difference at 1d postoperatively(P<0.05), there was no significant difference at 7d, 1, 3 and 6mo(P>0.05).Compared the aA and bA of ERG of the DR group before and after photocoagulation to the data of the HC group, there was a significant difference(P<0.05).Compared the aA and bA of ERG of the DR group after photocoagulation with the data before, there was a significant difference(P<0.05).Compared the aT and bT of ERG of the DR group before and after photocoagulation to the data of the HC group, there was a significant difference(P<0.05);compared the aT and bT of ERG of the group DR after photocoagulation with the data before, there was no significant difference(P>0.05).The vision of 49 cases(98 eyes) was improved 1 to 3 rows, the effective rate was 82%.The vision of 11 cases(22 eyes) remained constant.CONCLUSION: Retinal laser photocoagulation is an effective treatment to diabetic retinopathy patients, which can significantly reduce the peak systolic velocity of the central retinal artery, improve and stabilize the condition, protect visual function in long-term clinical effect.Color doppler flow imaging can observe the changes of ocular vessel flow velocity in diabetic` eyes noninvasively, repeatedly and in real time, providing a basis for clinical treatments.

OBJECTIVETo evaluate the application of shear wave elastography (SWE) combined with transition zone biopsy in the detection of prostate cancer (PCa).

METHODSA total of 489 patients with suspected PCa underwent transrectal ultrasonography (TRUS) and SWE-guided prostatic biopsy. We evaluated the role of SWE combined with transition zone biopsy in promoting the detection rate in comparison with the results of biopsy pathology.

RESULTSThe pathological results confirmed 221 malignant and 268 benign cases. Based on systematic biopsy, SWE combined with transition zone biopsy achieved a detection rate of 45. 19% , significantly higher than that of systematic biopsy alone (33.13%) (P < 0.05). The diagnostic sensitivity, specificity, and accuracy of SWE were significantly better than those of TRUS (P < 0.05). The mean elasticity (Emean) of SWE was remarkably higher for malignant than for benign lesions ([40.1 ± 9.5] vs [21.6 ± 8.3] kPa, P < 0.05). With 28.5 kPa as the threshold of the Emean value, the area under the ROC curve was 0. 899, and the diagnostic sensitivity and specificity were 88.71% and 86.23%, respectively.

CONCLUSIONSWE combined with transition zone biopsy could significantly improve the detection rate of prostate cancer.

Elasticity Imaging Techniques ; methods ; Humans ; Image-Guided Biopsy ; methods ; Male ; Prostate ; pathology ; Prostatic Neoplasms ; diagnosis ; diagnostic imaging ; pathology ; ROC Curve ; Sensitivity and Specificity

Objective To screen and clone the genes related to alopecia areata. Methods Dermal papillae of lesional and non lesional follicles were separated from alopecia areata scalp respectively. Suppression subtractive hybridization was used to investigate the difference of expressed genes in dermal papillae of lesional (tester) and non lesional (driver) follicles, and the differentially expressed genes in dermal papillae of lesional follicles were cloned and sequenced. Results A subtractive library of dermal papillae of lesional follicles from alopecia areata was established. A differentially expressed gene in dermal papillae of lesional follicles was successfully cloned and proved to be an autoantigen gene. Conclusions The subtractive library may contain the differentially genes related to alopecia. The autoantigen gene related to alopecia areata need to be further investigated.

Objectives To construct a cDNA subtractive library of dermal papilla cells (DPCs) in anagen with suppression subtractive hybridization (SSH) and clone differentially expressed genes related to DPCs in anagen. Methods Total RNA was isolated from DPC of anagen and telogen follicles. Then ds cDNAs were synthesized in turn using SMART cDNA synthesis technique. After cDNAs from anagen and telogen follicle DPCs were hybridized with each other twice and underwent two rounds of nested PCR, PCR products were ligated with arms of T/A plasmid vectors to set up the subtractive library. Selected clones were verified by reverse Nothern blot and DNA sequencing, and the acquired sequences were analyzed for homology based on Genbank nucleotide database. Results cDNA subtractive library of DPCs in anagen follicle was set up successfully with high subtractive efficiency. Thirty-five genes were identified with 22 known functional genes and 13 unknown functional genes. Conclusions These results demonstrate the effectiveness and sensitivity of SSH in detecting differentially expressed genes from a small amount of clinical samples. Information about such alterations in gene expression might be useful for elucidating the genetic events in hair follicle growth regulation.

Objective To investigate the influence of factors related to metabolic syndrome(MS)on the outcome in subjects without diabetes mellitus in a community.Methods A two-year follow-up study was conducted in 885 subjects who were enrolled in the epidemiologic survey carried out in Pingliang Community, Shanghai in 2002.Oral glucose tolerance test,lipid prefde,blood pressure(BP),body mass index(BMI),waist and hip circumferences were measured.Results (1)The baseline of BMI,fasting plasma glucose(FPG),2h plasma glucose after glucose loading(2hPG),BP,triglyceride(TG)in the subjects with impaired glucose regulation(IGR)increased significantly as compared to those with normal glucose regulation(NGR)(all P

Myeloid-derived suppressor cells (MDSCs) play a crucial role in T cell dysfunction, which is related to poor outcome in patients with severe trauma. Cyclooxygenase-2 (Cox-2) contributes to immune disorder in trauma and infection via production of prostaglandin E2. However, the role of Cox-2 in the accumulation and function of MDSCs after traumatic stress has not been fully elucidated. In the present study, we treated murine trauma model with NS398, a selective Cox-2 inhibitor. Then the percentages of CD11b+/Gr-1+ cells, proliferation and apoptosis of CD4+ T cells were determined. Arginase activity and arginase-1 (Arg-1) protein expression of splenic CD11b+/Gr-1+ cells, and delayed-type hypersensitivity (DTH) response were analyzed. The results showed that Cox-2 blockade significantly decreased the percentages of CD11b+/Gr-1+ cells in the spleen and bone marrow 48 and 72 h after traumatic stress. NS398 inhibited arginase activity and down-regulated the Arg-1 expression of splenic CD11b+/Gr-1+ cells. Moreover, NS398 could promote proliferation and inhibit apoptosis of CD4+ T cells. It also restored DTH response of traumatic mice. Taken together, our data revealed that Cox-2 might play a pivotal role in the accumulation and function of MDSC after traumatic stress.