Objective To construct programmed cell death protein-ligand 1(PD-LI)^hi tolerogenic dendritic cell (tol-DC) derived from bone marrow of DA rats and identify its immunological function. Methods DA rat bone marrow cells were extracted, combined with recombinant mouse granulocyte macrophage colony-stimulating factor and recombinant mouse interleukin (IL)-4, and cultured for 6 days in vitro to induce the differentiation of bone marrow cells into immature dendritic cells (imDC). Lipopolysaccharide was used to stimulate cell maturation and cultured for 2 days to collect mature dendritic cells (mDC). PD-L1 lentiviral vector virus stock solution or equivalent dose lentiviral stock solution was added, and PD-L1^hitol-DC and Lv-imDC were collected after culture for 2 days. The morphology of PD-L1^hitol-DC was observed by inverted phase contrast microscope and transmission electron microscope. Real-time fluorescence quantitative reverse transcription polymerase chain reaction, Western blotting and flow cytometry were used to detect the expression level of specific markers on cell surface. CD8⁺T cells derived from Lewis rat spleen were co-cultured with imDC, mDC, Lv-imDC and PD-L1^hitol-DC, respectively. The levels of inflammatory factors in the supernatant of each group were detected by enzyme-linked immunosorbent assay. The apoptosis of T cells and the differentiation of regulatory T cells (Treg) in each group were analyzed by flow cytometry. Results The morphology of PD-L1^hitol-DC modified by PD-L1 gene was consistent with tol-DC characteristics, and the expression levels of CD80, CD86 and major histocompatibility complex (MHC) on the surface were low. After mixed culture with CD8⁺ T cells, the levels of IL-10 and transforming growth factor (TGF) -β1 in the supernatant of PD-L1^hitol-DC group were higher, the levels of tumor necrosis factor (TNF) -α and IL-17A were lower, and the apoptosis of T cells and Treg differentiation were increased. Conclusions Overexpression of PD-L1 through lentiviral vectors may successfully induce the construction of bone-marrow derived PD-L1^hitol-DC in DA rats, promote the secretion of anti-inflammatory factors and T cell apoptosis, induce the differentiation of Treg, and inhibit the immune response of allogeneic CD8⁺T cells, which provides experimental basis for the next organ transplantation immune tolerance study.

Objective To explore the diagnostic value of a combined model based on clinicopathological and MRI features in BRCA-mutated ovarian cancer.Methods The data of 132 patients with ovarian cancer who underwent pathology and BRCA gene tes-ting were analyzed retrospectively,including 52 cases of BRCA mutation group and 80 cases of BRCA wild group.The differences of MRI features and clinicopathological features between BRCA mutation group and BRCA wild group were compared.Binary logistic regression was used to construct a joint prediction model and analyze its diagnostic efficiency.Results There were significant differ-ences in cytokeratin 7(CK7),estrogen receptor(ER),Ki-67 and lymphovascular invasion(LVI)between the BRAC mutation group and the BRAC wild group(P＜0.05).Univariate analysis showed that CK7,ER,Ki-67,LVI and the apparent diffusion coefficient(ADC)value of the cystic part of tumor were risk factors for BRCA-mutated ovarian cancer.The combined model based on CK7,ER,Ki-67,LVI,and the ADC value of the cystic part of tumor for the diagnosis of BRCA-mutated ovarian cancer had an area under the curve(AUC)of 0.765.Conclusion CK7,ER,Ki-67,LVI and the ADC value of the cystic part of tumor are risk factors for BRCA-mutated ovarian cancer.The combined model based on the above characteristics demonstrates good diagnostic efficacy for BRCA-mutated ovarian cancer.

Objective To explore the diagnostic value of a combined model based on clinicopathological and MRI features in BRCA-mutated ovarian cancer.Methods The data of 132 patients with ovarian cancer who underwent pathology and BRCA gene tes-ting were analyzed retrospectively,including 52 cases of BRCA mutation group and 80 cases of BRCA wild group.The differences of MRI features and clinicopathological features between BRCA mutation group and BRCA wild group were compared.Binary logistic regression was used to construct a joint prediction model and analyze its diagnostic efficiency.Results There were significant differ-ences in cytokeratin 7(CK7),estrogen receptor(ER),Ki-67 and lymphovascular invasion(LVI)between the BRAC mutation group and the BRAC wild group(P＜0.05).Univariate analysis showed that CK7,ER,Ki-67,LVI and the apparent diffusion coefficient(ADC)value of the cystic part of tumor were risk factors for BRCA-mutated ovarian cancer.The combined model based on CK7,ER,Ki-67,LVI,and the ADC value of the cystic part of tumor for the diagnosis of BRCA-mutated ovarian cancer had an area under the curve(AUC)of 0.765.Conclusion CK7,ER,Ki-67,LVI and the ADC value of the cystic part of tumor are risk factors for BRCA-mutated ovarian cancer.The combined model based on the above characteristics demonstrates good diagnostic efficacy for BRCA-mutated ovarian cancer.

Objective To observe the value of dual-source CT(DECT)virtual monoenergetic imaging(VMI)for differentiating prostate cancer(PC)and benign prostatic hyperplasia(BPH).Methods Thirty-three patients with PC(PC group)and 44 patients with BPH(BPH group)were retrospectively enrolled,and 40-100 keV(with 10 keV interval)VMI were reconstructed based on arterial phase DECT images,respectively.Clinical data,focal CT value and contrast-to-noise ratio(CNR)of VMI with different energy levels and conventional linear fusion images of arterial phase were compared between groups.Binary logistic regression models were constructed based on clinical data being significantly different between groups,focal CT value of VMI with energy level with the highest CNR,also focal CT value of conventional linear fusion images of arterial phase,respectively.The area under the receiver operating characteristic curve(AUC)was calculated to evaluate the efficacy of each model for differentiating PC and BPH.Results Free prostate-specific antigen(f-PSA),total prostate-specific antigen(t-PSA),focal CT value of 40-100 keV VMI and conventional linear fusion images of arterial phase in PC group were all higher,while short diameter in PC group was smaller than those in BPH group(all P＜0.05).Logistic regression models were constructed based on f-PSA,t-PSA,short diameter,CT value at 40 keV VMI and CT value at conventional linear fusion images of arterial phase,respectively,with AUC for differentiating PC and BPH of 0.879,0.902,0.701,0.911 and 0.857,respectively.Conclusion DECT VMI could be used as a supplementary examination for prostate diseases,and 40 keV VMI had the best efficacy for differentiating PC and BPH.

Objective: This study aims to analyze factors associated with lymphovascular space invasion (LVSI) and evaluate the prognostic significance of LVSI in Chinese endometrioid endometrial cancer (EEC) patients. Methods: Five-hundred eighty-four EEC patients undergoing surgery in our center from 2006 to 2016 were selected for analysis. Univariate analysis and multivariate logistic regression were used to examine relevant factors of LVSI. To evaluate the prognostic role of LVSI, survival analyses were conducted. In survival analyses, both multivariate Cox regression and propensity score matching were used to control the confounders. Results: The incidence of LVSI was 12.16% (71/584). Diabetes history (p=0.021), lymph node metastasis (p=0.005), deep myometrial invasion (p<0.001) and negative PR expression (p=0.007) were independently associated with LVSI. Both Kaplan-Meier method and univariate Cox regressions showed LVSI negative and positive cases had similar tumor-specific survival (TSS) and disease-free survival (DFS). After adjusting for the influence of adjuvant therapy and other clinicopathological factors with multivariate Cox regressions, LVSI still could not bring additional survival risk to the patients (p=0.280 and p=0.650 for TSS and DFS, respectively). This result was verified by Kaplan-Meier survival analyses after propensity score matching (p=0.234 and p=0.765 for TSS and DFS, respectively). Conclusion LVSI does not significantly compromise the survival outcome of Chinese EEC patients.

Objective To observe and compare the resistance changes of ciprofloxacin (CIP), levofloxacin (LEV), norfloxacin (NOR) to Pseudomonas aeruginosa (PA), and choose the drug which had the best curative effect for PA from the drugs mentioned above.Methods Seven strain of non-repetitive PA which had sensitiveness to ciprofloxacin (CIP), levofloxacin (LEV), norfloxacin (NOR) and the same minimal inhibitory concentration (MIC) of the three drugs were co-cultured with CIP, LEV, NOR in concentrations of 0.5, 1.0, 2.0, and 4.0 times of MIC for evoked drug resistance, respectively.The MIC of CIP, LEV, and NOR to PA which evoked drug resistance by 3 kinds of concentration were detected by the methods of Epsilometer Test and Two-fold Agar Dilution.Results The MIC value of CIP to PA after evoking by 3 concentrations increased from 0.09 μg/ml before evoking to 0.25-16.00 μg/ml.The MIC after evoking was 3 to 178 times that before evoking.The MIC value of NOR to PA after evoking increased from 0.50 μg/ml before evoking to 2.00-64.00 μg/ml.The MIC after evoking was 4 to 128 times that before evoking.The MIC value of LEV to PA after evoking by 3 concentrations increased from 0.50 μg/ml before evoking to 2.00-32.00 μg/ml.The MIC after evoking was 2 to 64 times that before evoking.The differences of MIC of CIP to PA after evoking by different concentrations of CIP, NOR and LEV were not statistically significant (all P>0.05).The MIC value of NOR to PA after evoking by 4.0×MIC CIP was higher than that of evoking by 0.5×MIC CIP (P=0.006).The MIC value of NOR to PA after evoking by 4.0×MIC LEV was higher than that evoked by 1.0×MIC LEP (P=0.006).The MIC value of LEV to PA after evoking by 4.0×MIC CIP was higher than that of evoking by 0.5×MIC CIP (P=0.006).The MIC value of LEV to PA after evoking by 4.0×MIC NOR and LEV were higher than those evoked by 0.5×MIC NOR and LEV (all P=0.007).Conclusion PA can be evoked to develop resistance to different degrees after co-culturing with different concentrations of CIP, LEV, and NOR.The change of resistance of CIP to PA is the least among the three drugs mentioned above.It is suggested that CIP is still one of the best fluoroquinolones for the treatment of PA infection.

Objective To observe and compare the resistance changes of ciprofloxacin (CIP), levofloxacin (LEV), norfloxacin (NOR) to Pseudomonas aeruginosa (PA), and choose the drug which had the best curative effect for PA from the drugs mentioned above.Methods Seven strain of non-repetitive PA which had sensitiveness to ciprofloxacin (CIP), levofloxacin (LEV), norfloxacin (NOR) and the same minimal inhibitory concentration (MIC) of the three drugs were co-cultured with CIP, LEV, NOR in concentrations of 0.5, 1.0, 2.0, and 4.0 times of MIC for evoked drug resistance, respectively.The MIC of CIP, LEV, and NOR to PA which evoked drug resistance by 3 kinds of concentration were detected by the methods of Epsilometer Test and Two-fold Agar Dilution.Results The MIC value of CIP to PA after evoking by 3 concentrations increased from 0.09 μg/ml before evoking to 0.25-16.00 μg/ml.The MIC after evoking was 3 to 178 times that before evoking.The MIC value of NOR to PA after evoking increased from 0.50 μg/ml before evoking to 2.00-64.00 μg/ml.The MIC after evoking was 4 to 128 times that before evoking.The MIC value of LEV to PA after evoking by 3 concentrations increased from 0.50 μg/ml before evoking to 2.00-32.00 μg/ml.The MIC after evoking was 2 to 64 times that before evoking.The differences of MIC of CIP to PA after evoking by different concentrations of CIP, NOR and LEV were not statistically significant (all P>0.05).The MIC value of NOR to PA after evoking by 4.0×MIC CIP was higher than that of evoking by 0.5×MIC CIP (P=0.006).The MIC value of NOR to PA after evoking by 4.0×MIC LEV was higher than that evoked by 1.0×MIC LEP (P=0.006).The MIC value of LEV to PA after evoking by 4.0×MIC CIP was higher than that of evoking by 0.5×MIC CIP (P=0.006).The MIC value of LEV to PA after evoking by 4.0×MIC NOR and LEV were higher than those evoked by 0.5×MIC NOR and LEV (all P=0.007).Conclusion PA can be evoked to develop resistance to different degrees after co-culturing with different concentrations of CIP, LEV, and NOR.The change of resistance of CIP to PA is the least among the three drugs mentioned above.It is suggested that CIP is still one of the best fluoroquinolones for the treatment of PA infection.

BACKGROUND:Previous studies have indicated that resistin stimulates a large set of chemokines in chondrocytes that are known to be important in inflammatory joint lesions.
OBJECTIVE:To further investigate the mechanism of co-regulation roles of transcription and post-transcription in the up-regulation of two chemokine genes CCL3 and CCL4 in chondrocytes in response to resistin.
METHODS:Human chondrocytes, T/C-28a2 and ATDC5 cels were cultured. The function of resistin on the chemokine genes, and the expression of C/EBPβ, nuclear factor-κB isoforms and chondrogenic specific miRNAs were tested by qPCR. The co-regulation of C/EBPβ and nuclear factor-κB was investigated by nuclear factor-κB inhibitor (IKK-NBD) and C/EBPβ inhibitor (SB303580) treatments, and subcelular localization was detected with or without resistin stimulation.
RESULTS AND CONCLUSION:Resistin could increase the expression of chemokine genes independently. Chondrocytes reacted in a non-restrictedly cel-specific manner to resistin; C/EBPβ inhibitor, nuclear factor-κB and some chondrogenic specific miRNAs in a combinatorial manner regulated chemokine gene expression. The activity of C/EBPβ was augmented by a transient increase in activity of nuclear factor-κB, and both transcription factors acted independently on the chemokine genes, CCL3 and CCL4.

BACKGROUND:The outcomes of SL-Plus? stem in primary total hip arthroplasty have been proved good, but whether SLR-Plus? revision stem can obtain a good outcome in revision hip arthroplasty needs more studies. OBJECTIVE:To observe the clinical results of revision hip arthroplasty with Zweymül er SLR-plus ? stem. METHODS:Revision hip arthroplasty of 41 hips in 39 patients was performed from November 1997 to May 2013 using SLR-plus ? stem. There were 26 hips (male) and 15 hips (female). They were at the age of 34-73 years old, 53 on average. In the 41 hips, 36 cemented and 5 uncemented femoral prostheses were used. Prosthetic changes on radiographs were observed. In accordance with Brooker classification, heterotopic ossification was classified to record the region and incidence of heterotopic ossification. Hip joint function was evaluated using Harris Hip Score, and survival rate of the prosthesis was analyzed. RESULTS AND CONCLUSION:A total of 31 patients (33 hips) were fol owed up for 1 to 16 years. The mean preoperative Harris hip score of 34 (range, 13-64) points improved to 85 (range, 55-94) points at the time of final fol ow-up. 32 hips (97%) had an excellent result. A 1 mm width radiolucent line was found in 1 femoral component without any symptom. Osteolysis and migration were seen in 1 hip, which needed re-revision. Heterotopic ossification developed in 9 hips, including 2 hips of Brooker grade 1, 4 hips of grade 2 and 3 hips of grade 3. No re-infection was found. Kaplan-Meier survivorship was 92%with radiographic loosening as the end point. Results demonstrated that the SLR-Plus? stem has sufficient immediate and long-term stability, which is reliable for patients undergoing hip revision surgery.

Objective To observe the distribution of circum-patella nerve fibers in the soft tissue to provide experimental evidence,which is significant in denervation for Total Knee Arthroplasty (TKA).Methods Patella specimens were collected from 4 cadavers (2 cases of diabetic foot,lcase of lower extremity arterial occlusive,and 1case of car accident),all 4 of which were resected soft tissue with a dimension of 0.5cm × 0.5cm and full depth thickness around patella more than 0.5cm for histology and morphology observation.The nerve fibers histology and morphology were observed in all resected specimens with HE staining and silver-gilt glycine staining in the same field of microscopic vision.Results Anatomy found that the vascular network form skins directly involved in the patella nourish hole area and in the 10,2,4,7 clock point have found that blood vessels into the patella.There have a large number of nerve fibers near to the patella under the microscope,but there were no significant difference in the nerve fibers region distribution of all specimens.There were some into patella nerve fiber paths in side of patella soft tissue,which lied in 7,11 and 13 clock point,but outside no this phenomenon.The distribution of circum-patellar nerve fibers were described as distribution of regional concentration,which lied in much more 5,6,7 clock points and 10,11,12,1,2,clock points,in which the quadriceps tendon and patellar tendon have more than the others.In the 13 clock point,the fascia and periosteum of nourish hole area were also found in a large number of nerve fibers,and there were laminar distribution in different soft tissue layers,which were collected much more in synovial layer,fat pad,tendon near to patella.Conclusion There are much more nerve fibers near to the patella and some into patella nerve fiber paths in the medial side and nourish hole area.Nerve fibers distribution of circum-patella can be described as laminar distribution and regional concentration ,which is more in the centre,bottom more than top,outside more than inside,the bipolar more than the others.The patella denervation operation by reducing the number of peripheral nociceptors to achieve desensitization is feasible in TKA.