Objective To study the mechanisms of inhibiting proliferous effect of human breast cancer cell line treated with genistein in vitro. Methods Two human breast cancer cell lines (MCF 7 and MDA MB 231) were cultured in vitro , proliferation of them was measured with MTT method, cell morphology was observed with Giemsa stain, cell cycle and apoptosis rate were measured with flow cytometry and apoptosis was detected in situ. Results Genistein markedly inhibited proliferation of MCF 7 and MDA MB 231 cell line. Typical image of apoptosis was observed in MCF 7 cell line treated with genistein in Giemsa stain:cell condensation, blebs formation on cell surface and pieces of condensed chromatin were scattered along nuclear envelope. The results detected with flow cytometry showed G 1 subpeak before G 1 phase peak (apoptosis peak). Apoptosis rate increased with time of genistein treatment. However, MDA MB 231 cell line treated with genistein did not show apoptosis and apoptosis peak, and quantity of cells at G 2 M phase obviously increased.Conclusion Genistein inhibites proliferation of human breast cancer cell line in vitro through inducing apoptosis or arresting cell at G 2 M phase.\;[

Objective To evaluate the efficacy and safety of Sun's tip-flexible ureterorenoscopy (tf-URS) combined with holmium laser lithotripsy in treatment of renal and upper ureteral calculi.MethodsClinical data of 48 patients (29 males and 19 females) with renal and upper ureteral calculi treated by tf-URS with holmium laser lithotripsy in our hospital from November 2015 to May 2016 were retrospectively analyzed.Among 48 patients there were 29 males and 19 females with a mean age of (37.8±12.5) years (21-67), including 31 cases of upper ureteral calculi and 17 cases of renal calculi with a mean stone size of (1.4±0.6) cm (0.8-2.1).The tf-URS was advanced through the ureter under visual direction with rigid mode.When the endoscope reached the ueteropelvic junction, the inner flexible part was extended out to explore the pelvicaliceal stones and then the holmium laser lithotripsy was performed.The therapeutic results were evaluated by KUB 2 days after operation and color ultrasonography or CT 4 weeks after operation.Results The success rate of insertion of tf-URS was 94% (45/48).The calculi were detected in 45 cases, and laser lithotripsy succeeded in 43 cases (96%).The rate of total stone clearance was 87% (39/45).The procedure was transformed to laparoscopic or percutaneous nephrolithotomy (PCNL) in 3 patients with ureterostenosis or ureteral distortion.No serious complications occurred.Conclusion The tf-URS with holmium laser in the treatment of renal and upper ureteral calculi is safe,reliable and effective.

Objective To detect the mouse testicular gene expression pattern differences be-tween spermatogonial stem cell (SSC) proliferative and differential stages and study the molecular reg-ulation mechanism in SSC proliferation and differentiation. Methods With the interval of 24 days, male Kunming mice were injected intraperitoneally with two doses of busulfan (10 mg/kg) to establish spermatogenesis regeneration models. 36 k Mouse Genome Array was used to detect the differential gene expression profiles between the stages of SSC proliferation and differentiation. Bioinforrnsties analysis was conducted in GO (gene ontology) and KEGG (Kyoto Encyclopedia of Genes and Ge-nomes) pathway to describe the potential roles that may play in spermatogonial stern cells behavior regulation. Results Nine hundred and eleven differential expression genes were identified by gene arrays in mice testes, consisting of 608 up-regulated and 303 down-regulated in SSC proliferation stage and SSC differentiation stage. The differential expression genes were classified by their biological process, molecular function and cellular component, respectively. Alterations with statistical signifi-cance (P<0.05)appeared in 84 KEG(;signal pathways, including Notch and Wnt signaling pathways which had been proved to be important for stem cell maintenance. Fifty-six differential expression genes were selected as genes related to stem cells, among which 40 genes were up-regulated, including some stem cell biomarkers(such as Cd9, StraS, hgbl-, Oct4 and Thyl)and some growth factors(such as Fgf2, Pdgfa and Csfl). Conclustion The regulation of SSC proliferation and differentiation involves inmany differentially expressed genes in various signal pathways. This study provides a molecular basis for the elucidation of the molecular mechanism behind self-renewal and differentiation of spermatogonial stem cells.

Objective To establish a long-term culture system for mouse spermatogonial stem cells(SSCs). Methods Testis cells from 4-6 days postpartum male transgenic BALB/C mice were collected by a modified two-step enzymatic digestion method.After three differential adherence selections,the enriched germ cells were finally suspended in StemPro-34 SFM medium supplemented with other nutrients factors and plated on mouse embryonic fibroblast(MEF)feeder layer.20 ng/ml Glial cell line-derived neurotrophic factor,10 ng/ml basic fibroblast growth factor and 200 ng/ml GDNF-family receptor al were added to the serum-free medium to promote SSCs proliferation.Aduh male BALB/C mice,4-5 weeks old,underwent intraperitoneal injection of 40 mg/kg busulfan as recipient mice.Cultured SSCs were also injected into the seminiferous tubules of the left recipient testis through micromanipulator and right testis as self-control.Testes of recipient mice were observed by a fluorescence stereomicroscope and HE stains at 2 months after transplantation. Results By improved digestion method,the vitality of isolated testis cells was more than 98%and the stem cells was enriched about 18.5 fold. 1-2 days after transferred to MEF feeder, the round germ cells started to proliferate and had the shape of paired or aligned undifferentiated spermatogonia connected by cytoplasmic bridges. After 3-4 days, SSCs proliferated continuously and formed typical colonies. SSCs from BALB/c mice could be cultured and passaged in a steady state for 3 months. Cryostat section through the transplanted testis showed that most of seminiferous tubules were filled with germ cells expressing EGFP.HE staining further showed clearly that seminiferous tubules contained complete spermatogenesis.Conclusions SSCs from BALB/c mice could be cultured in an improved culture system for 3 months.The culture system could facilitate understanding the regulatory mechanism that governs SSCs and might provide an opportunity for the cure of infertility.

ObjectiveTo study the value of the exfoliative cytology of prostatic fluid obtained from patients with elevated PSA in prostate cancer diagnosis.MethodsProstatic fluid was obtained from 130 patients with elevated PSA before prostate biopsy and then the Wright's stain and cytological class were done.Each cytological class and patient's age,PSA,total prostate volume,prostatic fluid volume and the number of leukocyte in the prostatic fluid were recorded.The relationship of leukocyte number and patient's age,PSA and prostate volume was analyzed by Spearman correlation test.ResultsProstate biopsy pathology results showed that there were 77 (59.2％) cancer cases and 53 (40.8％) non-cancer cases.Patient numbers in cytological class 1 to 5 were 28 (21.5％),32 (24.6％),22 (16.9％),36 (27.7％),12(9.2％),respectively.The prostate fluid cytology had a specificity of 100％ and high sensitivity of 62.5％(10/16) in patients with PSA≥20 μg/L.PSA value had significant difference between class 1,2,3,4and 5.Significant correlation was found among the prostatic fluid volume,total leukocyte number and prostate volume.Prostate volume,leukycyte density and total leukycyte number was significant higher in noncancer patients than in prostate cancer patients.ConclusionsThe exfoliative cytology of prostatic fluid is a valuable method in detecting prostate cancer,particularly in patients with high PSA levels.It has the advantages of non-invasion and less injury than prostate biopsy.There is a relationship between elevated PSA value and high leukocyte numbers.

OBJECTIVE:To determine the therapeutic effect of transpedicular instrumentation without fusion on patients with thoracolumbar burst fractures.METHODS:A total of 63 patients with thoracolumbar burst fractures (the inclusion criteria was neurologically intact spine with a kyphotic angle >20° and/or decreased anterior vertebral body height > 50%) who were treated with transpedicular instrumentation without fusion were studied,including 40 cases treated by AF internal fixation,16 cases by Tennor screw-rod fixation system and 7 cases by Diapason screw-rod fixation.All patients underwent a radiological and clinical assessment (including the loss of kyphotic angle,decreased anterior vertebral body height,the midsagital diameter of the canal and the Low Back Outcome Score) preoperatively,postoperatively and after 24 months.The deformity of angulation was measured by Cobb angle.RESULTS:All pstients were followed for a 24 months,with average stay of 13.4 days.There were averaged 3.8 days from admitted to operation,and the internal fixation was removed within 8-12 months in 51 cases,followed a 9.4-day hospital stay.According to low back outcome score,46 patients achieved excellent,9 good,5 fair and 3 poor,with excellent and good rates of 88%.The Cobb's angle was 20.1° preoperatively,6.2° postoperatively,and 11.9° after 24 months.The average lose of anterior vertebral body height was changed from 49.1% preoperatively to 17.4% postoperatively,which was 20.4% after 24 months.The midsagittal diameters was 49.8% (n=63) preoperatively,78.1% (n=28) postoperatively,and 91.7% (n=25) after 24 months.The implant failure occurred in 5 patients.The radiographic parameters had no associativity to the outcome of LBOS.CONCLUSION:Transpedicular instrumentation without fusion is conductive to treating burst fractures of the thoracolumbar spine without nerve injury.The routine posterior or posterolateral fusion is unnecessary in the operative management of these fractures.

Objective To observe CT features of primary yolk sac tumor (YST).Methods Clinical data and CT findings of 31 patients with primary YST proved by pathology were analyzed retrospectively.Plain CT was performed in 31 patients,while contrast enhanced CT scanning was performed in 23 patients.Results The lesions in 19 patients located in the gonads,including ovaries (n=11) and testes (n=8).Other lesions in 12 patients located out of gonads,including sacrococcygeal region (n =7),anterior mediastinum (n =3) and vagina (n=2).The tumors were oval shaped in 20 patients,while irregular shaped in other 11 patients.Well-defined boundary was found in 20 patients,whereas ill-defined boundary was found in 11 patients.Fat and calcification were found in 2 patients with teratomas.Moderate to marked enhancement of the solid part of tumors were observed in 23 patients,loofahs enhancement were observed in 17 patients,the blood vessels were found in 18 patients,while delayed enhancement of coated edge was found in 21 patients.The rupture of tumor capsule was found in 4 patients.Conclusion CT manifestations of YST have certain characteristics,which can provide imaging diagnostic evidences.

OBJECTIVETo investigate the mechanism of immune escape in renal cell carcinoma(RCC).

METHODSFas and FasL expressions were examined by immunohistochemical technique in 44 RCC patients, with the Ki67 expression and apoptosis of tumor infiltrating lymphocytes(TIL) monitored simultaneously. Cytokines including IL2 and IFN alpha were used to induce the expression of the renal carcinoma cell lines 786-0 cells. Combination treatment of 786-0 with cytokines and Anti-Fas monoclonal antibody (FasAb) was used to induce apoptosis. FasL function was assessed by in vitro co-culture assays using renal cancer cells 786-0 and Fas-sensitive Jurkat T-cells.

RESULTS(1) Fas expression rate in RCC(22.8%) was lower than that in the controlled normal kidney tissues(53.8%, P < 0.01). FasL expression rate in RCC (46.5%) was higher than that in the controlled normal kidney tissues (23.2%, P < 0.01). That of Ki67 was 32.8%, with the expressions of Fas and Ki67 showing a negative correlation (r = -0.62, P < 0.05). In contrast, the expressions of FasL and Ki67 showed a positive correlation. (r = 0.93, P < 0.01). The Fas expression of stage I was significantly higher than that of stages III and IV. The expression rate of FasL in RCC was significantly increased with RCC stage (P < 0.01). (2) The apoptotic rate of TIL in RCC (33.9%) was significantly higher than that of the normal kidney tissues (3.5%, P < 0.01). The expression of FasL and the apoptotic rate of TIL in RCC gave a positive correlation (r = 0.96, P < 0.01). (3) Fas expression rate in 786-0 cells was 13.7%. The apoptotic rate mediated by FAsAb was 9.6%. IFN alpha was able to up-regulate the Fas expression and subsequently augment the FasAb-mediated apoptosis in 786-0 cells. But IL2 did not show similar effects. (4) The FasL expression rate of 786-0 was 18.6%. FasL expressed by 786-0 cells was able to induce apoptosis of Jurkat T-cells in co-culture assays and the apoptosis of Jurkat T-cells was significantly lowered after blocking the effect of FasL with Fas-neutralizing antibody NOK-2, giving the apoptotic rates of 14.9% and 2.0%, respectively, the difference therein is statistically significant (P < 0.01).

CONCLUSIONDown-regulation of Fas expression and up-regulation of FasL-expression are the mechanisms through which the RCC cells escape from immune attack.

Adult ; Aged ; Carcinoma, Renal Cell ; immunology ; Fas Ligand Protein ; Female ; Humans ; Ki-67 Antigen ; immunology ; Kidney Neoplasms ; immunology ; Male ; Membrane Glycoproteins ; immunology ; Middle Aged ; fas Receptor ; immunology

Objective:The aim of this study was to study the characteristics of neoplasm invasion type in ovary of two orthotopic models established with human epithelial ovarian cancer solid tumor tissue slices and human ovarian carcinoma cell line OVCAR-3 in nude mice and human epithelial ovarian cancer.Methods:Tumor tissues and cell line OVCAR-3 of human epithelial ovarian cancer were grown in subcutaneous tissue and the subcutaneous tumor source was fetched and inoculated in ovarian capsule of nude mice to establish the orthotopic implantation model. The neoplasm invasion type in the two kinds of models were observed. The neoplasm invasion types were also analyzed by pathological examination in 54 cases of International Federation of Gynecology and Obstetrics (FIGO) stageⅠ-Ⅱepithelial ovarian cancer.Results:Three neoplasm invasion types were found as follows: type of pseudocapsule, type of pseudocapsule invasion, type of pseudocapsule penetration. Pseudocapsule rate in the solid tumor slices group (18.2%) were lower than those in the cell line group (42.3%) ( P<0.05), while the pseudocapsule penetration rate in the solid tumor slices (50.0%) were higher than those in the cell line group (23.1%) ( P<0.05). No difference was found of pseudocapsule invasion rate between two groups ( P>0.05). Neoplasm invasion type in ovary changed with tumor planting time. High proportion of pseudocapsule type was found at the beginning of tumor planting, and the pseudocapsule penetration rate raised with tumor planting time increased. High proportion of pseudocapsule type was also found in patients with FIGO stageⅠepithelial ovarian cancer, and pseudocapsule penetration rate increased in those with FIGO stageⅡ. No difference in neoplasm invasion type was found between two kinds of pathological types ( P>0.05). Conclusions:There are differences between the two kinds of orthotopic models established with human epithelial ovarian cancer solid tumor tissue slices and human ovarian carcinoma cell line OVCAR-3. Compared to the solid tumor slices model, the cell line model is more stable for the follow-up study. The proportion of three neoplasm invasion types in ovary were more balanced in 8 weeks after tumor planting, and 8 weeks after tumor planting is the best start time for the follow-up experiment.

Objective To investigate the value of modified YOLO-V5 model for identifying inflammatory bowel disease(IBD)displayed on CT enterography(CTE).Methods Totally 192 patients with IBD(103 cases of Crohn disease[CD subgroup]and 89 cases of ulcerative colitis[UC subgroup])and 103 patients with clinically suspected IBD but CTE showed no abnormality(no abnormality subgroup)were retrospectively collected as study group,while 5 patients with CD and 3 with UC were collected as test group.CTE images with diseased intestinal tubes present as thickened intestinal wall or no abnormality intestinal tubes were selected as data set(n=3 511).CTE in study group were divided into training set(n=3 160,including 1 063 from CD subgroup,931 from UC subgroup and 1 166 from no abnormality subgroup)and verification set(n=351,including 118 from CD subgroup,103 from UC subgroup and 130 from no abnormality subgroup)at the ratio of 9∶1,while 25 CET images(17 from 5 cases of CD and 8 from 3 cases of UC)in test group were used as test set.Diseased tubes of CD,UC and no abnormality tubes were labeled.Then 5 sub-models,including YOLO-V5n,YOLO-V5s,YOLO-V5m,YOLO-V5l and YOLO-V5x were constructed and trained with modified YOLO-V5,and their efficacy were verified in test set.Precision(Pr),recall(Rc)and mean average precision(mAP)were used to evaluate the efficacy of each sub-model for identifying IBD lesions displayed on CTE.Results The complexity of the above 5 sub-models increased successively.YOLO-V5l and YOLO-V5x sub-model had better diagnostic efficacy,the overall Pr,Rc,mAP_0.5 and mAP_0.5.0.95 of the former for identifying IBD lesions in training and validation sets was 0.97,0.93,0.96 and 0.91,while of the latter was 0.97,0.95,0.96 and 0.92,respectively.In test set,the efficacy of YOLO-V5n sub-model for identifying IBD lesions was low,with mAP_0.5∶0.95 of 0.66 and AUC of 0.82,whereas mAP_0.5∶0.95 of YOLO-V5x sub-model for identifying CD was as high as 0.92,and of YOLO-V5l sub-model for identifying UC was as high as 0.91.Conclusion YOLO-V5l and YOLO-V5x sub-models based on modified YOLO-V5 could effectively identify IBD lesions displayed on CTE.