Objective To investigate the change of apoptosis in LNCaP cells after inhibition of autophagic process under androgen removal conditions. Methods The autophagic level was deter-mined by using confocal microscopy and RT-PCR. The DAPI staining was used to indicate the apopto-sis of LNCaP cells after inhibition of autophagic by 3-MA. Also, Z-VAD-FMK was used to extend the apoptosis results. Results ①Androgen deprivation led to increased autophagy in LNCaP cells. LN-CaP cells cultured in complete medium(CM) presented low autophagic process with 1.9 scores. After 24 hours, the punetate GFP-LC3 structures were accumulated in the cells cultured in serum-free medi-um (SF)(2.64 scores). In contrast, the number of punctate GFP-LC3 remained at a very low level (1.85 scores), when cells were incubated with DHT in SFA(serum-free medium+DHT). Statistical analysis showed the significant difference between SF and SFA (P<0.01). Semiquantitative RT-PCR was employed to examine the mRNA expression of LC3. Indeed, cells grown in the medium without serum had a higher LC3 mRNA expression with the highest at 12 hour time point as compared with the cells grown in CM. DHT treatment reduced the level of LC3 mRNA. ②Blockage of autophagy by 3-MA increased the apoptosis of LNCaP cells. LNCaP cells in SF and SFA just presented a basal level of apoptosis, which is (3.19±1.09)% and (3.01±0.33)% , respectively. Under androgen-free con-ditions, inhibition of autophagy by 3-MA could increase apoptosis significantly(10. 90±2.91%). While Z-VAD-FMK, a pan Caspase inhibitor, was able to suppress this apoptotic process to the level of (1.16±0.52)%, which was statistically significant(P<0.01). Conclusions Androgen removal can lead to the increase of autophagy in LNCaP cells. Moreover, inhibition of autophagy promotes the occurrence of apoptosis.

Objective To investigate the long-term effects of propofol postconditioning on cerebral ischemia-reperfusion (I/R) injury in rats.Methods One hundred and forty-four healthy male SD rats,aged 7-8 weeks,weighing 250-280 g,were equally and randomly divided into 4 groups:sham operation group (group S),I/R group,propofol postconditioning group (group P) and intralipid group (group I).The animals were anesthetized with intraperitoneal 10％ chloral hydrate 300 mg/kg.Focal cerebral ischemia was induced by occlusion of middle cerebral artery for 60 min using a nylon thread with a rounded tip which was inserted into internal carotid artery in groups I/R,P and I.Two hour infusion of propofol was started at 20 mg· kg- 1· h- 1 immediately after the onset of reperfusion in group P,while the equal volume of normal saline was given instead in S and I/R groups,and 10％ intralipid was given instead in group I.Five rats in each group were chosen on day 1,14 and 28 after operation for assessment of neurological behavior and detection of cerebral infarct volume.Six rats in each group were chosen to perform Morris water maze test at day 9 and 23 after operation for 6 consecutive days.Five rats in each group were sacrificed on day 1,14 and 28 after operation and the hippocampal tissues were isolated for determination of the expression of GluR1-containing AMPA (GluR1-AMPA) receptor and GluR1-AMPA receptor in cell membrane.The ratio of GluR1-AMPA receptor in cell membrane/GluR1-AMPA receptor was calculated.Results Compared with group S,neurological behavior scores and the number of animals' swimming across the platform were significantly decreased,cerebral infarct volume was significantly enlarged,escape latency was significantly prolonged,and ratio of GluR1-AMPA receptor in cell membrane/GluR1-AMPA receptor was significantly increased ( P ＜ 0.05),while no significant change in the expression of GluR1-AMPA receptor was found in I/R group ( P ＞0.05).Propofol postconditioning inhibited cerebral I/R-induced changes mentioned above ( P ＜ 0.05).Conclusion The brain protection against focal I/R injury by propofol postconditioning can last for 28 days after operation and the inhibition of trafficking of GluR1-AMPA receptor from cytoplasm to cell membrane may contribute to this long-term brain protection.

Objective To identify the risk factors for prostate-sparing cystectomy by evaluating the risk of prostatic invasion or incidental prostatic adenocarcinoma (PCa) in bladder cancer (BCa) patients undergoing radical cystectomy.Methods The patients undergoing radical cystectomy from 2009 to 2014 in Fujian Medical University Union Hospital were enrolled to analyze the risk factors of prostatic tumor invasion.These factors included age,tumor size,location,quantity,histologic grade and pathologic stage.Results In the 123 male patients,the mean age was 60 years (range,31-78 years);23 (18.7％) patients had BCa or PCa in the prostate;14 (11.4％) had prostatic Bca;11 (8.9％) had PCa.The risk factors of prostatic BCa included multifocal bladder tumors (OR =26.70,P =0.032),tumor in the bladder neck and trigone(OR =17.13,P =0.013),pathological stage (OR =26.70,P ＜ 0.001).Among the 11 patients with PCa,3(27.3％) patients had Gleason score of ≥7,8(72.7％) patients ≤6 and 2(18.2％) patients had extracapsular extension.Three patients had clinically significant PCa.The factor of advanced age was associated with incidental PCa (P =0.003).Conclusion The risk factors of prostatic tumor invasion in patients undergoing radical cystectomy included advanced age,bladder tumor in bladder neck and trigone,muhifocal bladder tumors,and advanced pathological stage.

Objective To study the effects of sensitized dendritic cells in the treatment of bladder tumor and further discuss the mechanism of this immunotherapy. Methods 44 female F344 rats, which irrigated N-methyl-N-nitrosourea into bladders every other week for a total of five doses, were induced to bladder tumor. They were treated subcutaneously with either PBS, unsensitized DC, freeze thawing supernatant of tumor cells, or sensitized DC respectively every week for a total of four times. In the fifteenth week, their bladders were weighted and harvested for observation by naked eye and microscope, their blood was harvested for examination CTL by FCM. Results The weight of bladders in sensitized DC group was lower than those in PBS group, unsensitized DC group and freeze thawing supernatant of bladder tumor cells group (P<0.05). The stages of bladder tumor in sensitized DC group were statistically descended compared with those in PBS group (P <0.05). The CD+3 T cells in sensitized DC group was lower than those in PBS group, unsensitized DC group and freeze thawing supernatant of bladder tumor cells group (P <0.05). The CD+3 CD+8 CD+28 T cells in sensitized DC group was higher than those in PBS group, unsensitized DC group and freeze thawing supematant of bladder tumor cells group(P <0.001). Conclusion Sensitized DC injecting subcutaneously can reduce the stages of F344 rats' bladder tumor, Unsensitized DC injecting subcutaneously has not effect in the treatment of bladder tumor;, while the effect of freeze thawing supematant of tumor cells injecting subcutaneously is not well. The mechanism of sensitized DC in the treatment of blader tumor is that DC plays an immunol killing role by presenting antigen, stimulating CTL.

Objective To compare the short-term efficacy and adverse effects of docetaxe or oxaliplatin combined with capecitabine in the treatment of late-staged gastric cancer in aged patients. Methods Eighty-two aged patients with late-staged gastric cancer were randomly divided into two groups,of which 38 patients were treated group) ,and 44 patients were treated with oxaliplatin (100 mg/m2 ivgtt on 1st day) and eapecitabine (2000 mg/1 cycle). Results There is no failure of follow-up. In the docetaxe group,the effective rate was 52.63% (20/38) and 54.55 % (24/44) for the docetaxe and oxaliplatin group,respectively (P>0.05). The median progression-free survival(PFS) in the docetaxe group (6.1 months) was similar to that in the oxaliplatin group (6.3 months) (P>0.05). Gastrointestinal response,myelosuppression and neurotoxicity (Ⅰ or Ⅱ level) were the most common ad-verse effects observed in both groups (P>0.05). No chemotherapy-related death was observed. Conclusions The short-term efficacy of decetaxe or oxaliplatin combined with capecitabine in the treatment of late-staged gastric cancer in aged patients is similar,and the adverse effects are all within tolerance limits.

Objective To predict the constitutive parameters of a superelastic model of plantar soft tissues based on random forest(RF)and backpropagation(BP)neural network algorithms to improve the efficiency and accuracy of the method for obtaining constitutive parameters.Methods First,a finite element model for a spherical indentation experiment of plantar soft tissues was established,and the spherical indentation experiment process was simulated to obtain a dataset of nonlinear displacement and indentation force,divided into training and testing sets.The established RF and BP neural network(BPNN)models were trained separately.The constitutive parameters of plantar soft tissues were predicted using experimental data.Finally,the mean square error(MSE)and coefficient of determination(R2)were introduced to evaluate the accuracy of the model prediction,and the effectiveness of the model was verified by comparison with the experimental curves.Results Combining the RF and BPNN models with finite element simulation was an effective and accurate method for determining the superelastic constitutive parameters of plantar soft tissues.After training,the MSE of the RF model reached 1.370 2×10-3,and R2 was 0.982 9,whereas the MSE of the BPNN model reached 4.858 1×10-5,and R2 was 0.999 3.The inverse-determined constitutive parameters of the plantar soft tissues suitable for simulation were obtained.The calculated response curves for the two predicted sets of constitutive parameters were in good agreement with the experimental curves.Conclusions The prediction accuracy for the superelastic constitutive parameters of plantar soft tissues based on an artificial intelligence algorithm model is high,and the relevant research results can be applied to study other mechanical properties of plantar soft tissues.This study provides a new method for obtaining the constitutive parameters of plantar soft tissues and helps to quickly diagnose clinical problems,such as plantar soft tissue lesions.

Objective To explore risk factors of the progression to castration-resistant prostate cancer(CRPC)after hormone therapy (HT).Methods A total of 178 patients with prostate cancer from February 2009 to February 2018 were enrolled to analyze the risk factors of the progression to castrationresistant prostate cancer after androgen deprivation therapy in Fujian Medical University Union Hospital.The mean age was72 years (range,49-91 years);the middle Gleason score was 7 (range,4-10);the middle PSA at the initiation of HT was 24.45 ng/ml (range,0.16-100.0 ng/ml);the middle time to PSA nadir was 9 months (range,0.5-69.0 months);the middle PSA nadir after HT was 0.030 ng/ml (range,0.003-78.670 ng/ml);the mean hemoglobin level was 131 g/L (range,64-184 g/L);the mean alkaline phosphatase level was 98 U/L (range,35-734 U/L);39 patients were diabetes mellitus (21.9％);82 patients were bone metastasis/visceral metastasis (46.1％);85 patients (47.8 ％) were in clinical T1 + T2;93 patients(52.2％)were in clinical T3 + T4.We studied the relationship between CRPC and these risk factors including age,Gleason score,PSA at the initiation of HT,PSA nadir after HT,the time to PSA nadir,hemoglobin level,alkaline phosphatase,bone metastasis/visceral metastasis,clinical T stage,diabetes mellitus by x2 test,univariate and multivariate Cox regression analysis methods.Results The middle follow-up time was 30 months (range,6-92 months).There were 74 of 178 patients progressed to CRPC after HT.The median time of progression to CRPC in this cohort was 15 months (range,4-47 months).On x2 test analysis,there were statistically significant differences between the progression to CRPC group after HT and the rest group in Gleason score (P ＜0.001),PSA nadir after HT (P ＜0.001),PSA at the initiation of HT (P =0.042),alkaline phosphatase (P =0.002),bone metastasis/visceral metastasis (P＜0.001) and clinical T stage (P ＜0.001).Additionally,on multivariate Cox regression analysis,Gleason score (OR =6.152,P ＜ 0.001),PSA nadir after HT (OR =3.022,P ＜ 0.004) and the time to PSA nadir (OR =0.375,P ＜0.001) were found to be significantly associated with the rapid progression to CRPC.Conelusions Gleason score,PSA nadir after HT and the time to PSA nadir were significantly associated with the progression to CRPC.Patients with higher PSA nadir or the shorter time to PSA nadir were more likely to progress to CRPC.

Objective:To explore the surgical treatment strategy of stent graft infection after interventional treatment of major iliac artery related diseases.Methods:Retrospective analysis was performed on the clinical data of 6 patients with secondary stent graft infection after interventional treatment for major iliac artery related diseases admitted to the Department of Vascular Surgery,Affiliated Drum Tower Hospital,Medical School of Nanjing University from November 2021 to August 2022.There were 5 males and 1 female,with a mean age of 64 years (range:49 to 79 years).The infection time was 53 days to 3 165 days.All the 6 patients received surgical treatment,including 3 patients who underwent anatomic bypass grafting (axillary arterial-femoral artery bypass,femoral arterial-femoral artery bypass) using artificial vessels,and 3 patients who underwent in situ abdominal aorta reconstruction using bovine pericardium.The perioperative situation,postoperative infection and the occurrence of serious adverse events were recorded,and the safety of different treatment methods and materials was evaluated.Results:All patients successfully completed the operation and no death occurred during hospitalization.Intraoperative blood loss was 2 000 to 5 000 ml,and intraoperative blood transfusion was 1 600 to 5 350 ml.All the patients were followed up for 81 to 395 days after surgery,and the incision healed well,and no reinfection occurred.Postoperative gastrointestinal bleeding occurred in 1 patient,secondary surgery (retroperitoneal hematoma removal) was performed in 1 patient due to postoperative bleeding at the vascular anastomosis,both lower limb amputations were performed in 1 patient due to postoperative lower limb ischemia,and intermittent claudication occurred in 2 patients.All patients were alive at the last follow-up.Conclusion:For patients with aortic stent graft infection,when the infection is not serious and there is enough space to block the proximal and distal aorta,in situ aortic reconstruction is an effective treatment,and different materials can achieve satisfactory results in a short period of time.

Objective:To explore the surgical treatment strategy of stent graft infection after interventional treatment of major iliac artery related diseases.Methods:Retrospective analysis was performed on the clinical data of 6 patients with secondary stent graft infection after interventional treatment for major iliac artery related diseases admitted to the Department of Vascular Surgery,Affiliated Drum Tower Hospital,Medical School of Nanjing University from November 2021 to August 2022.There were 5 males and 1 female,with a mean age of 64 years (range:49 to 79 years).The infection time was 53 days to 3 165 days.All the 6 patients received surgical treatment,including 3 patients who underwent anatomic bypass grafting (axillary arterial-femoral artery bypass,femoral arterial-femoral artery bypass) using artificial vessels,and 3 patients who underwent in situ abdominal aorta reconstruction using bovine pericardium.The perioperative situation,postoperative infection and the occurrence of serious adverse events were recorded,and the safety of different treatment methods and materials was evaluated.Results:All patients successfully completed the operation and no death occurred during hospitalization.Intraoperative blood loss was 2 000 to 5 000 ml,and intraoperative blood transfusion was 1 600 to 5 350 ml.All the patients were followed up for 81 to 395 days after surgery,and the incision healed well,and no reinfection occurred.Postoperative gastrointestinal bleeding occurred in 1 patient,secondary surgery (retroperitoneal hematoma removal) was performed in 1 patient due to postoperative bleeding at the vascular anastomosis,both lower limb amputations were performed in 1 patient due to postoperative lower limb ischemia,and intermittent claudication occurred in 2 patients.All patients were alive at the last follow-up.Conclusion:For patients with aortic stent graft infection,when the infection is not serious and there is enough space to block the proximal and distal aorta,in situ aortic reconstruction is an effective treatment,and different materials can achieve satisfactory results in a short period of time.

Human beings are inevitably exposed to toxic substances as a result of influences of potential contamination factors in the environment,food and medicines,which poses a threat to human health.In order to effectively screen and prevent the exposure or intake of such substances,it is neces-sary to develop in vitro assays for the detection and toxicity evaluation of toxic substances.High content screening(HCS)has been recognized as an important tool for toxicity testing and risk assessment of compounds due to its high throughput and automation advantages,and has been widely used in in vitro toxicology research.In this review,we described the system components of HCS and its workflow in toxicity screening and toxicity evaluation by focusing on cases of their application in toxicity detection and evaluation studies,including the cytotoxicity,hepatotoxicity,nephrotoxicity,genotoxicity,neurotox-icity,cardiotoxicity,and developmental toxicity.In addition,the applications and developments of machine learning in HCS were explored,especially to the advantages of supervised and unsupervised machine learning strategies for high throughput image screening and data analysis.Finally,the future applications of HCS in toxicity screening and evaluation are outlined,especially in terms of binding new models and gene editing technology.