Background::Bladder cancer, characterized by a high potential of tumor recurrence, has high lifelong monitoring and treatment costs. To date, tumor cells with intrinsic softness have been identified to function as cancer stem cells in several cancer types. Nonetheless, the existence of soft tumor cells in bladder tumors remains elusive. Thus, our study aimed to develop a microbarrier microfluidic chip to efficiently isolate deformable tumor cells from distinct types of bladder cancer cells.Methods::The stiffness of bladder cancer cells was determined by atomic force microscopy (AFM). The modified microfluidic chip was utilized to separate soft cells, and the 3D Matrigel culture system was to maintain the softness of tumor cells. Expression patterns of integrin β8 (ITGB8), protein kinase B (AKT), and mammalian target of rapamycin (mTOR) were determined by Western blotting. Double immunostaining was conducted to examine the interaction between F-actin and tripartite motif containing 59 (TRIM59). The stem-cell-like characteristics of soft cells were explored by colony formation assay and in vivo studies upon xenografted tumor models. Results::Using our newly designed microfluidic approach, we identified a small fraction of soft tumor cells in bladder cancer cells. More importantly, the existence of soft tumor cells was confirmed in clinical human bladder cancer specimens, in which the number of soft tumor cells was associated with tumor relapse. Furthermore, we demonstrated that the biomechanical stimuli arising from 3D Matrigel activated the F-actin/ITGB8/TRIM59/AKT/mTOR/glycolysis pathways to enhance the softness and tumorigenic capacity of tumor cells. Simultaneously, we detected a remarkable up-regulation in ITGB8, TRIM59, and phospho-AKT in clinical bladder recurrent tumors compared with their non-recurrent counterparts.Conclusions::The ITGB8/TRIM59/AKT/mTOR/glycolysis axis plays a crucial role in modulating tumor softness and stemness. Meanwhile, the soft tumor cells become more sensitive to chemotherapy after stiffening, that offers new insights for hampering tumor progression and recurrence.

Objective:To investigate the efficacy of the biopsy strategy combining 6-core systematic and 3-core MRI-targeted biopsy on prostate cancer (PCa) detection in biopsy-na?ve patients.Methods:The clinical data of 121 biopsy-na?ve patients who underwent transperineal prostate biopsy in West China Hospital of Sichuan University from July 2018 to January 2020 were retrospectively analyzed. The average age was (64.7±9.1) years old. Pre-biopsy prostate-specific antigen (PSA) was (12.4±7.5)ng/ml, f/t PSA was 0.13±0.05. Prostate volume was (43.1±26.1) ml and PASD was (0.35±0.27) ng/ml 2. The prostate-imaging and data system (PI-RADS) score of MRI before biopsy was reported to be 3 for 29 patients (24.0%), 4 for 54 patients (44.6%) and 5 for 38 patients (31.8%). All 121 patients underwent 12-core systematic biopsy combined with a 3-core or 5-core MRI-targeted biopsy, of which 61 patients underwent 3-core targeted biopsy and 60 underwent 5-core targeted biopsy. There was no significant difference in the pre-biopsy clinical data between the two groups ( P>0.05). A 6-core systematic biopsy was redefined as the results of 6 cores among the 12-core systematic biopsy. We compared the detection rates among the single 12-core systematic biopsy, 6-core systematic biopsy, MRI-targeted biopsy (3-core or 5-core), and different systematic biopsy combing with targeted biopsy for any PCa and clinically significant PCa, and we also analyzed the cumulative cancer detection rates for MRI-targeted biopsy of different cores. Results:Of the 121 patients in this study, the biopsy results were negative for 43 patients (35.5%) and positive for 78 (64.5%). The detection rate of clinically significant PCa was 55.4% (67/121). The detection rate of the 6-core systematic biopsy combined with MRI-targeted biopsy was 62.0% (75/121) for PCa and 55.4% (67/121) for clinically significant PCa, which was of no difference compared with that for the 12-core systematic biopsy combined with MRI-targeted biopsy ( P>0.05), but the 6-core systematic biopsy combined with MRI-targeted biopsy avoided the overdiagnosis of 3 patients with Gleason score 3+ 3. The detection rate of PCa for MRI-targeted biopsy was 57.9% (70/121), including 42.1% (51/121) for the first core, 55.4% (67/121) for the first two cores, and 57.9% (70/121) for the first three cores. Compared with the single-core targeted biopsy for suspicious lesions, the first 2-core targeted biopsy ( OR=1.7, 95% CI 1.0-2.8) and 3-core targeted biopsy ( OR=1.9, 95% CI 1.1-3.1) can significantly increase the detection rate of PCa, while the fourth or fifth core of targeted biopsy can not increase the detection rate additionally (60%, 36/60). Conclusion:For patients with suspected PCa, the prostate biopsy strategy combing 6-core systematic and 3-core MRI-targeted biopsy performs no inferior than the current 12-core systematic biopsy combined with MRI-targeted biopsy.

Objective : To observe the effect of hydrogen sulfide( H2 S) on the expression of transforming growth factor⁃β1(TGF⁃ β1) , E ⁃cadherin (E⁃CAD) , Vimentin (VIM) , alpha⁃smooth muscle actin ( α ⁃SMA) during epithelial⁃mesenchymal transformation(EMT) , and to explore the anti⁃fibrosis mechanism of it. Methods : Sixty rats ( male SD) were randomly divided into control group, bleomycin group, NaHS + bleomycin group and prednisolone + bleomycin group , 15 rats per group. 5 rats of each group were sacrificed at random in 7th , 14th and 28th day. The degree of alveolitis and pulmonary fibrosis was observed . The expression of protein and mRNA of TGF⁃ β1 , E ⁃cad , VIM , α ⁃SMA were determined by Immunohistochemi stry and RT⁃PCR. Results : ① HE and Masson staining showed that the lung tissue of fibrosis had the lowest degree in control group. and the most severe in bleomycin group. The lung tissue of NaHS + bleomycin group and prednisolone + bleomycin group also had alveolitis and fibrosis changes , but the degree Were significantly less than bleomycin group. ② The mRNA and protein expression levels of TGF⁃ β1 , VIM and α ⁃SMA in bleomycin group, NaHS + bleomycin group and prednisolone + bleomycin group were all higher than that in control group at 7th , 14th and 28th day ( P < 0. 05 ) , while the expression levels of them in NaHS + bleomycin group and prednisolone + bleomycin group were all lower than that in bleomycin groupat each time (P < 0. 05) , which was significant at 28th day in NaHS + bleomycin. ③ The mRNA and protein expression levels of E ⁃Cad in bleomycin group, NaHS + bleomycin group and prednisolone + bleomycin group were all lower than that in control group at 7th , 14th and 28th day(P < 0. 05) , but the expression levels of E ⁃Cad in⁃NaHS + bleomycin group and prednisolone + bleomycin group were higher than that in bleomycin group at each time(P < 0. 05) , which was significant at 28th day in NaHS + bleomycin. Conclusion H2 S can reduce the degree of pulmonary fibrosis in rats , its mechanism may be related to the down⁃regulation of TGF⁃ β1and the inhibition of the EMT , which can enhance the expression of E ⁃cad and reduce the expression of TGF⁃ β1 , VIM and α ⁃SMA.

With the rapid development of organ donation and transplantation, it becomes increasingly important to improve the management system and ensure the quality and safety of organs for transplantation. Guide to the Quality and Safety of Organs for Transplantation (6th Edition) by European Committee provides the definitions of severe adverse reactions and severe adverse events in the perspective of biosafety early warning, delivers management and reporting processes, and encourages medical staff involved in organ transplantation to identify the adverse reactions and adverse events as soon as possible, and to carry out investigation, assessment and feedback. At the same time, it also systematically illustrates the warning and monitoring of organ transplantation risk, which is worthy of application in clinical study and practice.

OBJECTIVE:To establish a method for the content determination of 8 metal elements in Propylgaclate and sodium chloride injection. METHODS:Microwave digestion-inductively coupled plasma mass spectrometry(ICP-MS) was adopted. Radiofrequency power was 1 530 W;cooling temperature was 4 ℃;collision gas was He gas;carrier gas was argon;flow rate of carrier gas was 1.08 L/min;integration time was 0.3 s;plasma gas flow rate was 15 L/min;the vacuum degree of quadrupole was 3.04×10－4 Pa;sampling cone aperture was 1.0 mm;interception cone aperture was 0.4 mm;the speed of sampling was 0.3 rps;data collection was repeated for 3 times. The microwave digestion power is 1 600 W,and the heating process is heated to 160℃at room temperature for 30 min,and maintained at 5 min,and then heated to 190 ℃ at a temperature of 5 ℃/min and maintained 45 min. RESULTS:The linear range of Mg and Al were 1-250 ng/mL;the linear range of Cr,Mn,Fe,Cu,Zn,Cd were 1-100 ng/ mL(all r≥0.999 0). The limits of detection were 0.063 6-1.785 0 ng/mL. RSDs of precision,stability and reproducibility tests were all lower than 4%. The recoveries were 89.65%-105.60%(RSD were 1.57%-3.98%,n＝9). RSDs of durablity were all lower than 12%. CONCLUSIONS:The method is simple,accurate,precise,stable,reproducible and durable. It can be used for content determination of 8 metal elements in Propylgaclate and sodium chloride injection.

Objective: To establish a method for the determination of eucalyptol, camphor and menthol in compound menthol camphor eucalyptus oil solution by GC.Methods: An HP-INNOWAX 19091N-216 capillary column(60 m× 0.32 mm , 0.50 μm)was used.The carrier gas was nitrogen with the flow rate of 30 ml·min-1 , the gas was hydrogen with the flow rate of 40 ml·min-1 and the oxidant gas was air with the flow rate of 400 ml· min-1.The detector was FID and the inlet temperature was 250℃.The temperature program was as follows: the initial column temperature was 50℃, and then risen to 160℃ with a rate of 10℃·min-1 and kept for 5 min, and finally risen to 220℃ with a rate of 20℃·min-1 and kept for 3 min.The split ratio was 15∶1 and the injection volume was 1 μl.Results: The linear range of eucalyptol, camphor and menthol was 0.031 9-2.550 0 mg·ml-1 (r=1.000 0), 0.041 3-3.305 0 mg·ml-1 (r=1.000 0) and 0.053 7-4.294 0 mg·ml-1 (r=1.000 0), respectively.The average recovery was 98.24% (RSD=0.3% , n =9), 98.97% (RSD=0.4% , n =9) and 98.98% (RSD=0.5% , n =9), respectively.Conclusion: The method is sensitive and accurate with good stability, which can be used to determine the contents of eucalyptol, camphor and menthol in compound menthol camphor eucalyptus oil solution.

Objective:To prepare monoclonal antibodies specifically against an immunogenic fragment in ectodomain of prostate-specific membrane antigen ( PSMA ).Methods: An polypeptide immunogenic fragment in the ectodomain of PSMA was predicted by biological information technology,and then it was expressed prokaryotically.BALB/c mice were immunized with the prokar-ytically expressed recombinant polypeptide antigen,to prepare the monoclonal antibodies specifically against an immunogenic fragment in ectodomain of PSMA by hybridoma technology,purification of monoclonal antibody by affinity chromatography,characterization of the monoclonal antibodies by Western blot.The radioimmunoimaging in prostate cancer model was performed by using the labeled McAb.Results:Throught the software analysis,we got the antigen fragment in the ectodomain of PSMA containing 310aa sequences higher specificity, artificially synthesized gene sequence of the region, and constructed a prokaryotic expression vector pET-32a-r-ectodomain-PSMA,by prokaryotic expression we obtained the 50 kD target antigen,after hybridization,the three positive hybridoma cell lines (5E6,4A5 and 4D7) were selected by ELISA using target antigen,the isotypes of 5E6 and 4A5 were IgG2a,the isotypes of 4D7 were IgG1,the titer of three monoclonal antibodies was above 1∶256 000.Western blot results showed that the prepared monoclonal anti-bodies could binding specifically to the antigen in the ectodomain of PSMA.Radioimmunoimaging in prostate cancer animal model results further confirm that the prepared monoclonal antibodies could combinate with the antigen in the ectodomain of PSMA in the animal body, and make the tumor imaging.Conclusion: The prepared monoclonal antibodies can specifically recognizes the PSMA antigen,which laid the foundation for the immunodiagnosis and immunotherapy of prostate cancer.

Objective To establish regression m odel betw een craniofacial lines and body height by m ea-suring craniofacial lines in Southw est H an m ales using C Tand to accum ulate data for the study of foren-sic anthropology. Methods H ead C Tdata of 273 H an m ales in Southw est w ere collected and 7 cranio-facial lines w ere determ ined. M ultiplanar reconstruction and volum e rendering w ere perform ed by im age post-processing softw are and the selected lines w ere m easured. The relationship betw een each m easuring indicator and body height w as analyzed using SPSS 21.0 softw are. The regression equation of body height estim ation w as established and 50 sam ples w ere selected again and put into the m athem atics m odels to verify its accuracy. Results The linear regression equations of 7 lines w ere established (P<0.05). The correlation coefficients of the unary linear regression equations w ere 0.190-0.439 and the standard errors of the estim ate (SEE) w ere 4.597-5.023 cm . The correlation coefficients of the m ultiple linear regression equation w ere 0.494-0.524 and the SEEw ere 4.418-4.458 cm . The return tests show ed that the highest ±1SEEaccuracy of the m ultiple regression equation:y=83.959+3.589 x6+2.573 x2, w ere 30%;and the highest ±2SEEaccuracy of the m ultiple regression equation: y=72.646+3.316 x6+1.586 x2+1.553 x4+2.211 x3, w ere 92% . Conclusion There is significant linear correlation betw een 7 selected lines and the stature in this study, and the plural linear regression equation established could be applied for estim ating the stature of Southw est H an m ales.

Objective To establish the linear regression equation between body height and com bined length of manubrium and mesosternum of sternum m easured by CTvolum e rendering technique (CT-VRT) in southw est H an population. Methods One hundred and sixty subjects, including 80 m ales and 80 fem ales w ere selected from southw est H an population for routine CT-VRT(reconstruction thickness 1 m m ) ex-am ination. The lengths of both manubrium and mesosternum w ere recorded, and the com bined length of manubrium and mesosternum was equal to the algebraic sum of them . The sex-specific linear regression equations between the com bined length of manubrium and mesosternum and the real body height of each subject w ere deduced. Results The sex-specific sim ple linear regression equations between the com bined length of manubrium and mesosternum (x3) and body height (y) w ere established (m ale:y=135.000+2.118x3 and fem ale:y=120.790+2.808x3).Both equations show ed statisticalsignificance (P<0.05) w ith a 100% predictive accuracy. Conclusion CT-VRTis an effective m ethod for m easurem ent of the index of sternum . The com bined length of manubrium and mesosternum from CT-VRTcan be used for body height estim ation in southw est H an population.

Objective To evaluate the roles of signal transducer and activator of transcription 3(STAT3) played in delayed xenograft rejection.Method Mice-to-rats cardiac xenograft model was established and recipients were administrated by inhibitor of phosphorylation of STAT3,5,15-Diphenylporphyrin(DPP),and immunosuppressive agent,ciclosporin A (CsA) alone or both.The survival of graft was analyzed.Immunohistochemistry and immunoblotting were utilized to detect the phosphorylated STAT3 (p-STAT3),and real-time polymerase chain reaction was used to assess STAT3-targeted genes in grafts.Result The expression of p-STAT3 was increased significantly with the prolonged survival in grafts (P ＜ 0.05).Administration of DPP and CsA both significantly prolonged survival of the grafts (P＜0.05),and decreased the expressions of STAT3-targeted genes including Bcl-xl,Bcl-2,Cyclin D1,C-myc and VEGF (P＜0.05).DPP and CsA exerted the synergistic effects.Conclusion STAT3,which is persistently activated in cardiac xenografts,probably up-regulates downstream genes to promote proliferation and to inhibit apoptosis of endothelial cells,leading to the activation of endothelial cells and delayed xenograft rejection.