Objective To describe the phenotype of NK cells in Bama miniature pigs, and establish an efficient activation and culture method for porcine cytokine-induced killer ( CIK) cells in vitro. Methods The porcine peripheral blood mononuclear cells ( PBMCs) were isolated by Percoll gradient centrifugation, and the phenotype of NK cells was test-ed by detecting the CD2 + /CD8 + /CD3 - cell compartment. To establish an efficient activation and culture method for por-cine CIK cells, we optimized the culture conditions to improve the CIK activation efficiency. Results Using the optimized induction culture conditions, the ratio of CIK ( CD2 + /CD8 + /CD3 -) cells was up to 43. 63% at the fifth day, approxi-mately 5. 59 times increased compared with the initially separated PBMCs. Cell proliferation experiments showed that three obvious fluorescence peaks were observed on the fifth day. The results indicated that the induced CIK cells underwent three times cell division, in theory, about increased 8-fold compared with the initial separation of PBMCs. Furthermore, the qRT-PCR result of the surface markers of porcine NK cells also showed a similar variation tendency as the flow cytometry results. Conclusions Our findings demonstrate the successful establishment of an efficient activation and culture method for porcine CIK cells in vitro.

Objective This study was to explore the expression and significance of HMA2 in bladder cancer , analyze its correlations to clinicopathologic and recurrence of bladder cancer. Methods The expression of HMGA2 protein in 148 specimens of bladder cancer and 30 specimens of normal bladder tissues was detected by immunohistochemtry, its correlations to clinicopathologic features was analyzed. Results There was no expression of HMGA2 protein in normal bladder tissues,while the expression level of HMGA2 protein was getting higher with the increase of tumor pathology grade and stage. The positive rate of HMGA2 protein was 21.3% in G1 bladder cancer, 60. 3% in G2 bladder cancer, 82.1% in G3 bladder cancer, its difference is significant (P＜0. 001). It was significantly lower in non-muscle invasive bladder cancer than in muscle invasive bladder cancer (43.3% vs 72. 7%, P=0. 003). The patients were followed up for 2～95 months, patients of recurrence was 64,HMGA2 protein expression was significantly higher in patients with recurrence than with non-recurrence (54.7% vs 25.0%, P=0. 007). Conclusions The expression of HMGA2 protein was highly in bladder cancer, the positive rate of HMGA2 protein expression was related with classification,TMN stage and recurrence, but not with sex, age, tumor number (P＞0. 05). The detection of the expression of HMGA2 protein is in favor of diagnosis and prognostic evaluation of bladder cancer.

Objective To investigate the value of imaging-pathology fusion guided targeted cryoablation for localized prostate cancer.Methods Between June 2014 and February 2017,64 patients undergoing targeted cryoablation for localized prostate cancer were retrospectively collected and analyzed.The average age was 77.8 years old,ranging 45-87 years old.The preoperative valne of PSA ranged from 5.1 to 19.8ng/ml,with mean of 15.3 ng/ml,including 6 case of cT1c,35 cases of cT2a,and 23 cases of cT2b,and 35 cases of Gleason score 6,8 cases of 3 + 4 and 21 cases of 4 + 3.All patients had good continence and 11 cases had good erectile function before operation.All the patients received MRI or contrast-enhanced ultrasonography examinations before biopsies,and the sites and range of cryoablation were determined by MRI or contrast-enhanced ultrasonography and biopsy pathological results.Results All the patients underwent the surgery successfully.The average postoperative hospital stay was 1.6 days,ranging 1 to 3 days,including 22 cases discharged within 24 h after surgery.The operative duration ranged from 85 to 152 min,with mean of 99.6 min.The satisfaction rate of the patients was 100％.Seven patients had Clavien Ⅰ or Ⅱ postoperative complications,and recovered after symptomatic treatment.The follow-up duration ranged from 0.5 to 32.2 months,with median of 11.8 months.All patients were alive without new metastasis.Eight patients developed biochemical recurrence,5 of those underwent intermittent endocrine therapy,resulting in stable PSA levels.Conclusion Imaging-pathology fusion guided targeted cryoablation was safe and effective for localized prostate cancer with the advantages of few complications,high satisfaction rate and quick recovery.

Objective To develop a real-time RT-PCR assay ( rRT-PCR) for efficient detection of duck hepatitis virus type 1 ( DHV-I) .Method According to the different gene sequences of DHV-I from different provinces download from NCBI and to find the conserved sequences.One pair of the specific primers and one TaqMan probe were designed. Then reaction parameters were optimized to develop a real-time RT-PCR assay ( rRT-PCR) .Results This developed rRT-PCR assay could detect 20 template copies of RNA, and its sensitivity was higher than that of the conventional RT-PCR. This rRT-PCR assay was found to be specific and able to detect DHV-I, and no positive results were observed when nucleic acid from Muscovy duck parvovirus, goose parvovims, Newcastle disease and avian influenza virus, egg drop syndrome virus, reticuloendotheliosis virus, duck Tembusu virus, poultry intestinal arc virus were used as rRT-PCR templates.The results of this developed rRT-PCR assay used for 100 duck clinical samples showed a positive rate of 92%, indicating that DHV exists in duck group of Jiangsu province in China.Conclusion This rRT-PCR assay can be used as a rapid tool for detection of DHV-I.

Objective· To evaluate the safety of neoadjuvant therapy which was constituted by docetaxel based systemic chemotherapy and maximal androgen blockage for patients with locally advanced prostate cancer and to summarize the related adverse events and clinical managements.Methods· From June 2015 to February 2017,the clinical data of 55 patients undergoing neoadjuvant chemotherapy combined with complete androgen deprivation were retrospectively reviewed.The patients were given docetaxel and prednisone as DP regimen every 3 weeks and LHRH analogues with bicalutamide as maximal androgen deprivation for a total of 4 cycles.All treatment-related adverse events were observed and then recorded.Results· Two cases with liver function impairment after 2 cycles of treatment were withdrawn from the study.No severe allergic reactions occurred during neoadjuvant therapy.The most common adverse events were hematologic toxicity,while 23.6％ of patients had grade Ⅲ-Ⅳ neutropenia,and about 12.7％ had anemia.Due to a relatively short course of treatment,the skin or mucous damage,peripheral neurotoxicity and fluid retention were rare.However,hot flash,male breast development as well as erectile dysfunction were very frequently observed due to maximal androgen deprivation.The majority of these adverse events were relieved by symptomatic and supportive treatment.Conclusion · After strict selection,4 cycles of neoadjuvant chemotherapy combined with total androgen blockade could be well tolerated by the patients with high-risk locally advanced prostate cancer.Even though the adverse events were controllable,they still need to be closely monitored during treatment in order to reduce the incidence.In addition,the very low testosterone level associated endocrinal metabolic disorders caused by complete androgen deprivation were also of great concern.

Objective To investigate genotyps of Treponema pallidum (Tp) in several cities in Guangxi province. Methods A total of 300 patients with suspected early syphilis were enrolled from STD clinics in Guangxi between January 2012 and July 2016, and tissue fluid samples were collected from skin lesions. Silver staining was performed to detect Tp, and PCR to amplify the Tp polA gene for the diagnosis of early syphilis. Positive samples were subjected to PCR amplification of a 60-bp tandem repeat region within the arp gene, restriction fragment length polymorphism(RFLP)analysis of the tpr Ⅱgene after digestion with Mse Ⅰ enzyme and tp0548 genotyping. Results Finally, 215 patients were diagnosed with early syphilis, including 210(97.7%)patients positive for PCR and 105(48.8%)patients positive for silver staining, and the positive rate significantly differed between the two methods (χ2 = 103.01, P < 0.05). Among the PCR-positive samples, 190 could be genotyped by analysis of three target genes, and 17 genotypes were identified. The genotype 14d/f was predominant (45.3%, 86/190), followed by 15d/f (13.7%, 26/190), 16d/f(11.6%, 22/190), 17d/f(7.4%, 14/190), 13d/f(6.8%, 13/190), 10d/f(4.2%, 8/190), 18d/f(1.6%, 3/190), 16a/f(1.6%, 3/190), 5d/f(1.1%, 2/190), 7d/f(1.1%, 2/190), 12d/f(1.1%, 2/190), 16d/e(1.1%, 2/190), 14a/f(1.1%, 2/190), 9h/c(1.1%, 2/190), 15l/f(0.5%, 1/190), 25a/e(0.5%, 1/190), 15i/f(0.5%, 1/190). Conclusion Tp genotypes are diversified in patients with early syphilis in Guangxi, and the genotype 14 d/f is predominant.

Objective To investigate the efficacy and safety of docetaxel chemotherapy combined with androgen-deprivation therapy (ADT) for patients with metastatic hormone-sensitive prostate cancer.Methods One hundred and ninety-two cases of metastatic hormone-sensitive prostate cancer in Renji Hospital between January 2015 and July 2016 were analyzed retrospectively.Patients' age was 39 to 90,the median age was 71 years.The median prostate-specific antigen (PSA) at diagnosis was 90.6ng/ml (4.1-2 556.0 ng/ml).One hundred and eighty were with bone metastasis and 12 were with distant lymphatic metastasis.Sixty-one of them received docetaxel chemotherapy plus ADT for 3 weeks,131 received hormonal treatment alone.The median age of combination therapy group was 67 years (39-80 years),that of single treatment group was 75 years (50-93 years) (P ＜ 0.001).The median PSA baseline of the two groups were 91.6 ng/ml (35.5-157.5ng/ml) and 89.1 ng/ml (59.6-191.0 ng/ml) (P =0.324).Gleason score of combination therapy group showed that 3 cases (4.9％) was 6,23 cases (37.7％) 7,35 cases (57.4％) ≥8.That of single treatment group showed that 17 cases (13.0％) 6,51 cases (38.9％) 7,63 cases (48.1％) ≥8.There was no statistic difference between the two groups (P =0.122).But there was statistic difference in the rate of T3 or T4 clinical stage in primary lesion,that of combination therapy group was 50.7％ (37/61) and 34.4％ (21/61),and that of single treatment group was 60.3％ (79/131) and 21.4％ (28/131) (P =0.011).Imaging showed local lymph node metastasis in the two groups (80.3％ vs.67.9％,P =0.005).As to physical condition,the combination therapy group showed a lower ECOG score than the single treatment group (P ＜ 0.001).All the patients' survival condition,PSA response rate and adverse events were analyzed.Results One hundred and ninety-two patients were regularly followedup.The median follow-up time was 23.3 (14.4-33.4) months.Median progression free survival time of combination therapy group and single treatment group were respectively 24.4 (7.5-31.3) months vs.17.5(3.0-30.7) months (P ＜ 0.001).There were 1 and 16 cases died in the two groups due to disease progression.During the treatment,the rate of PSA level less than 0.2 ng/ml was 29.5％ (18/61) vs.13.7％ (18/131) in combination therapy group and single treatment group.Regarding the tolerance of combination therapy group,the incidence rate of grade 3-4 neutropenia was 27.9％ (17/61).Skin and mucous membrane damaged in 24.6％ (15/61) patients,transaminase rised in 13.1％ (8/61) patients,and peripheral nerve toxicity occurred in 9.8％ (6/61) patients.There was no significant difference between the 2 groups in relevant events caused by ADT,gynecomastia (14.8％ vs.16.3％) and erectile dysfunction (100％ vs.100％).Most of them could be relieved by symptomatic treatment.Conclusions For metastatic hormone-sensitive prostate cancer,docetaxel combined with hormonal treatment showed longer progression free survival than ADT alone with adverse reactions acceptable.

Objective To evaluate the predictive value of CT scan in the outcome of renal function after decompression procedures in patients with acute obstructive renal failure.Methods Retrospective analysis of clinical information of 76 patients with acute obstructive renal failure between January 2012 and August 2017 was performed.Clinical information included age,cause of obstruction,type of decompression procedures,total renal area and dominant side renal area measured by CT scan,serum creatinine(SCr) level before operation,SCr level in 3 months after decompression.Patients were divided into 2 groups according to creatinine clearance rate (CCr) in 3 months after decompression,including normal renal function group (26 cases,CCr ≥50 ml/min) and decompensated renal function group (50 cases,CCr ＜ 50 ml/min).The CCr of normal renal function group was (65.3 ± 12.8)ml/min,while the decompensated renal function group was (28.9 ± 11.9)ml/min.The average age of normal renal function group was 48.3 years old,the median value of serum creatinine before operation was 633μmol/L,while the average age of decompensated group was 63.8 years old,the median value of preoperative creatinine was 583 μmol/L.Renal area was measured in CT scan image which was performed when acute obstructive renal failure was diagnosed.Univariate analysis and logistic regression multivariate analysis was used to analyze these parameters.Receiver operating characteristic curve (ROC) was used to evaluate the performance of CT scan for renal function prediction.Results There were significant differences in the renal area between the normal renal function and decompensated renal function group,the total renal area were (3 765.5 ± 628.6) mm2 (range 2 375.3-4 853.6 mm2) and (2 493.0 ± 830.6) mm2 (range 476.9-4 225.1 mm2) respectively (P ＜ 0.001),while the dominant side renal area were (2 283.5 ± 430.2) mm2 (range 1 654.6-3 383.3 mm2) and (1 655.5 ± 496.2) mm2 (range 476.94-2 816.0 mm2) respectively (P ＜ 0.001).The integration area under the ROC curve of the total renal area to predict the outcome of renal function was 0.89.Conclusions The renal area measured in CT scan image might be useful for the early prediction of renal function outcome in patients with acute obstructive renal failure.

Objective To investigate the efficacy of pretreatment systemic inflammation response index (SIRI) in predicting the prognostic of prostate cancer (PCa) patients treated with maximal androgen blockade (MAB).Methods The data of 351 PCa patients who had undergone MAB as first-line therapy between January 2010 and June 2015,were retrospectively analyzed.The age of patients in our cohort ranged from 51 to 89 years old,mean 76 years old.The median value of PSA was 91.60ng/ml,ranging 0.11-1 000.00 ng/ml.39 cases had a Gleason score of 6,47 cases had a score of 3 + 4,89 cases had a score of 4 +3,107 cases had a score of 8,and 69 cases had a score of 9-10.158 cases had bone metastasis.Patients were categorized in two groups using a cut-off point of 1.2 as calculated by the receiver-operating curve analysis.Correlations between SIRI and clinical characteristics were analyzed.Meanwhile,univariate and multivariate cox regression analyses were performed to determine the associations of SIRI with progression-free survival (PFS),cancer-specific survival (CSS) and overall survival (OS).Results The median follow-up duration was 43.0 months,ranging 9-75 months.The disease progression occurred in 162 patients,91 patients died,including 75 who died because of PCa at the end of the last follow-up.The differences of age,Gleason score and incidence of metastasis between low SIRI group (＜ 1.2) and high SIRI group (≥1.2) were not significant (P ＞0.05).But the patients in high SIRI group had higher PSA (P =0.046).Multivariate analyses identified SIRI,Gleason score and metastasis as independent prognostic factors for PFS,CSS and OS.Conclusions Pretreatment SIRI ≥ 1.2 was an independent predictor for poor prognosis in PCa patients treated with MAB.

Purpose: In non-metastatic prostate cancer (nmPCa) setting, it is important to early identify the patients at risk of biochemical recurrence (BCR) for immediate postoperative intervention. Our study aimed to evaluate the potential clinical utility of circulating tumor DNA (ctDNA) for predicting disease recurrence. Materials and Methods: This real-world observational study evaluated 161 cases of nmPCa undergoing next-generation sequencing at our institution. A total of 139 ctDNA samples and 31 biopsied tumor tissue underwent genomic profiling. The study endpoint was BCR after radical prostatectomy. Relationships between the ctDNA status and the biochemical progression-free survival (bPFS) were analyzed by log-rank test and multivariate Cox regression. Results: Of 161 enrolled patients, 19 (11.8%) harbored deleterious alterations in NCOR2, followed by BRCA2 (3.7%), ATR (2.5%), and CDK12 (2.5%). Of available pre-operative blood samples (n=139), ctDNA was detectable in 91 (65.5%). Until last follow-up, 56 of 68 patients (85.3%) with detectable ctDNA had achieved BCR, whereas only eight of 39 patients (20.5%) with undetectable ctDNA had achieved BCR. Patients who had undetectable ctDNA experienced significantly longer bPFS compared with those who had detectable ctDNA (not available vs. 8.2 months; hazard ratio, 0.14; p < 0.01). Pre-operative ctDNA status was a significant prognostic factor of disease recurrence. Conclusion Pre-operative ctDNA detection could identify patients at high risk of recurrence and has the potential to inform immediate postoperative interventions, but these approaches remain to be validated in prospective studies. ctDNA studies can provide insights into accurate monitoring and precise treatment rather than simply following routine clinical care.