1.Identification of biomarkers associated with ferroptosis and pyroptosis for the potential diagnosis of postmenopausal osteoporosis
Shudong LI ; Xuezhen LIANG ; Di LUO ; Jiacheng LI ; Bozhao YAN ; Gang LI
Chinese Journal of Tissue Engineering Research 2024;28(28):4511-4515
		                        		
		                        			
		                        			BACKGROUND:Ferroptosis and pyroptosis may play a role in the development of postmenopausal osteoporosis.There may be relevant biomarkers for the diagnosis of postmenopausal osteoporosis. OBJECTIVE:To search for the key genes related to ferroptosis and pyroptosis in postmenopausal osteoporosis using bioinformatics so as to further elucidate their biological mechanisms. METHODS:The data sets GSE56815 and GSE7429 of postmenopausal osteoporosis were downloaded from the GEO database,the national comprehensive gene expression database of the United States,and the two data sets were preprocessed.The differential expression analysis of the data was carried out by the limma package of R software,and the enrichment analysis was performed by DIVID and KOBAS.The protein-protein interaction network was mapped by STRING and Cytoscape,the Hub gene was selected by CytoHubba,and the key genes were screened by the ferroptosis database and pyroptosis database.The CIBERSORT package was used to determine the immune infiltration of postmenopausal osteoporosis samples and to analyze the correlation between key genes and immune cells RESULTS AND CONCLUSION:A total of 30 differential genes of postmenopausal osteoporosis were screened in the experimental samples,of which 9 genes were up-regulated and 21 genes were down-regulated.The enrichment of GO and KEGG pathways showed that the differences were mainly in"serine-type endopeptidase activity,""innate immune response,""special particle lumen,"and"renin secretion."The protein-protein interaction network showed the correlation of differential genes and the top 10 Hub genes with"Degree"value were selected using CytoHubba.Hub gene was intersected with the FerrDb database and cell pyroptosis dataset to obtain key genes ELANE and LCN2.Receiver operating characteristic curve and box diagram showed that the expression of ELANE and LCN2 in serum samples of postmenopausal osteoporosis was significantly lower than that in normal samples,indicating a good diagnostic value.Immune infiltration analysis showed that ELANE may be related to memory resting CD4+ T cells,M0 and M2 macrophages.LCN2 may be related to M0 macrophages.
		                        		
		                        		
		                        		
		                        	
2.Prognostic factors of patients with muscle invasive bladder cancer with intermediate-to-high risk prostate cancer
Junyong OU ; Kunming NI ; Lulin MA ; Guoliang WANG ; Ye YAN ; Bin YANG ; Gengwu LI ; Haodong SONG ; Min LU ; Jianfei YE ; Shudong ZHANG
Journal of Peking University(Health Sciences) 2024;56(4):582-588
		                        		
		                        			
		                        			Objective:To investigate the prognostic factors for all-cause mortality in patients with muscle-invasive bladder cancer(MIBC)with intermediate-to-high-risk primary prostate cancer.Methods:From January 2012 to October 2023,the clinical data of the patients with MIBC with intermediate-to-high-risk primary prostate cancer in Peking University Third Hospital were retrospectively analyzed.All the patients were monitored and the occurrence of all-cause death was documented as the outcome event in the prognostic study.Univariate and multivariate Cox proportional risk regression analysis models were implemented to search for independent influences on the prognosis of patients.For significant influencing factors(pathological T stage,M stage and perineural invasion of bladder cancer),survival curves were plotted before and after multifactorial Cox regression adjusting for confounding factors.Results:A total of 32 patients were included in this study.The mean age was(72.5±6.6)years;the median preoperative total prostate specific antigen(tPSA)was 6.68(2.47,6.84)μg/L;the mean preoperative creatinine was(95±36)μmol/L,and the median survival time was 65 months.The majority of the patients(87.5%)had high-grade bladder cancer,53.1%had lymphatic invasion,and 31.3%had perineural invasion.Prostate involvement was observed in 25.0%of the cases,and the positive rate of soft-tissue surgical margin was 37.5%.Multivariate Cox analysis revealed that preoperative creatinine level(HR=1.02,95%CI:1.01-1.04),pathological stage of bladder cancer T3(HR=11.58,95%CI:1.38-97.36)and T4(HR=19.53,95%CI:4.26-89.52)metastasis of bladder cancer(HR=9.44,95%CI:1.26-70.49)and perineural invasion of bladder cancer(HR=6.26,95%CI:1.39-28.27)were independent prognostic factors(P<0.05).Survival curves with Log-rank test after adjusting for confounding factors demonstrated that bladder cancer pathology T3,T4,M1,and perineural invasion were unfavorable factors affecting the patients'survival prognosis(P<0.05).Conclusion:Patients with MIBC with intermediate-to-high risk primary prostate cancer generally portends a poor prognosis.High preoperative serum creatinine,T3 or T4 pathological stage of bladder cancer,metastasis of bladder cancer and bladder cancer perineural invasion are poor prognostic factors for patients with MIBC with intermediate-to-high risk primary prostate cancer.
		                        		
		                        		
		                        		
		                        	
3.Clinicopathological characteristics and prognosis of multilocular cystic renal neoplasm of low malignant potential
Le YU ; Shaohui DENG ; Fan ZHANG ; Ye YAN ; Jianfei YE ; Shudong ZHANG
Journal of Peking University(Health Sciences) 2024;56(4):661-666
		                        		
		                        			
		                        			Objective:To analyze the clinicopathological characteristics and prognosis of patients with multilocular cystic renal neoplasm of low malignant potential and compare the clinicopathological charac-teristics of patients with multilocular cystic renal neoplasm of low malignant potential who underwent different surgical methods.Methods:Clinicopathological data and prognosis of patients admitted to Peking University Third Hospital from January 2010 to September 2023 were collected.Patients who underwent radical nephrectomy or nephron-sparing surgery and were pathologically diagnosed with multilocular cystic renal neoplasm of low malignant potential were identified.Based on the surgical methods,the patients were divided into radical nephrectomy group and nephron-sparing surgery group.The clinicopathological characteristics of the two groups were compared.Results:A total of 35 patients were enrolled in this study.The median age at diagnosis was 53.0(39.0-62.0)years.Among the 35 patients,23 were males(65.7%)and 12 were females(34.3%).Nine patients underwent radical nephrectomy(25.7%),while 26 patients underwent nephron-sparing surgery(74.3%).The clinical T-stage of 35 patients did not exceed T2a stage.The median operation time was 145.0 min,and the median estimated intraoperative blood loss was 20.0 mL.The median postoperative hospitalization days was 6.0 d.The postoperative pathological results did not indicate renal sinus invasion,sarcomatous change,adrenal invasion or lymph node invasion.Based on the surgical methods,the patients were divided into a radical nephrectomy group and a nephron-sparing surgery group.There was no significant difference in clinicopathological charac-teristics between the two groups.Except for one patient who was lost to the follow-up,all the other patients were followed up for 8-111 months,with a median follow-up time of 70.5 months.Only one patient died from non-cancer-specific reasons,other patients had no tumor metastasis or recurrence.Conclusion:Patients with multilocular cystic renal neoplasm of low malignant potential have a good prog-nosis.There is no significant difference in clinicopathological characteristics of patients between nephron-sparing surgery group and radical nephrectomy group for multilocular cystic renal neoplasm of low malig-nant potential.
		                        		
		                        		
		                        		
		                        	
4.Predicting the 3-year tumor-specific survival in patients with T3a non-metastatic renal cell carcinoma
Zezhen ZHOU ; Shaohui DENG ; Ye YAN ; Fan ZHANG ; Yichang HAO ; Liyuan GE ; Hongxian ZHANG ; Guo-Liang WANG ; Shudong ZHANG
Journal of Peking University(Health Sciences) 2024;56(4):673-679
		                        		
		                        			
		                        			Objective:To predict the 3-year cancer-specific survival(CSS)of patients with non-meta-static T3a renal cell carcinoma after surgery.Methods:A total of 336 patients with pathologically con-firmed T3a N0-1M0 renal cell carcinoma(RCC)who underwent surgical treatment at the Department of Urology,Peking University Third Hospital from March 2013 to February 2021 were retrospectively collect-ed.The patients were randomly divided into a training cohort of 268 cases and an internal validation co-hort of 68 cases at an 4∶1 ratio.Using two-way Lasso regression,variables were selected to construct a nomogram for predicting the 3-year cancer-specific survival(CSS)of the patients with T3aN0-1M0 RCC.Performance assessment of the nomogram included evaluation of discrimination and calibration ability,as well as clinical utility using measures such as the concordance index(C-index),time-dependent area un-der the receiver operating characteristic curve[time-dependent area under the curve(AUC)],calibra-tion curve,and decision curve analysis(DCA).Risk stratification was determined based on the nomo-gram scores,and Kaplan-Meier survival analysis and Log-rank tests were employed to compare progres-sion-free survival(PFS)and cancer-specific survival(CSS)among the patients in the different risk groups.Results:Based on the Lasso regression screening results,the nomogram was constructed with five variables:tumor maximum diameter,histological grading,sarcomatoid differentiation,T3a feature,and lymph node metastasis.The baseline data of the training and validation sets showed no statistical differences(P>0.05).The consistency indices of the column diagram were found to be 0.808(0.708-0.907)and 0.903(0.838-0.969)for the training and internal validation sets,respectively.The AUC values for 3-year cancer-specific survival were 0.843(0.725-0.961)and 0.923(0.844-1.002)for the two sets.Calibration curves of both sets demonstrated a high level of consistency between the actual CSS and predicted probability.The decision curve analysis(DCA)curves indicated that the column dia-gram had a favorable net benefit in clinical practice.A total of 336 patients were included in the study,with 35 cancer-specific deaths and 69 postoperative recurrences.According to the line chart,the patients were divided into low-risk group(scoring 0-117)and high-risk group(scoring 119-284).Within the low-risk group,there were 16 tumor-specific deaths out of 282 cases and 36 postoperative recurrences out of 282 cases.In the high-risk group,there were 19 tumor-specific deaths out of 54 cases and 33 post-operative recurrences out of 54 cases.There were significant differences in progression-free survival(PFS)and cancer-specific survival(CSS)between the low-risk and high-risk groups(P<0.000 1).Conclusion:A nomogram model predicting the 3-year CSS of non-metastatic T3a renal cell carcinoma patients was successfully constructed and validated in this study.This nomogram can assist clinicians in accurately assessing the long-term prognosis of such patients.
		                        		
		                        		
		                        		
		                        	
5.Retrospective study on the impact of penile corpus cavernosum injection test on pe-nile vascular function
Yan CHEN ; Kuangmeng LI ; Kai HONG ; Shudong ZHANG ; Jianxing CHENG ; Zhongjie ZHENG ; Wenhao TANG ; Lianming ZHAO ; Haitao ZHANG ; Hui JIANG ; Haocheng LIN
Journal of Peking University(Health Sciences) 2024;56(4):680-686
		                        		
		                        			
		                        			Objective:To investigate the impact of age,various hormonal levels,and biochemical markers on penile cavernous body vascular function in patients with erectile dysfunction(ED).Me-thods:A retrospective analysis of clinical data from male patients with ED who underwent color duplex Doppler ultrasonography(CDDU)and intracavernosal injection test(ICI)at the Reproductive Medicine Center of Peking University Third Hospital from January 2020 to August 2023.Data were managed and processed using SPSS 29.0,and a multivariable Logistic regression analysis was conducted.Results:A total of 700 ED patients were included,with 380 showing negative ICI results and 320 positive.In the study,84 patients had a peak systolic velocity(PSV)<25 cm/s,while 616 had PSV ≥ 25 cm/s;202 patients had end-diastolic velocity(EDV)>5 cm/s,and 498 had EDV ≤5 cm/s.264 patients had ab-normal PSV and/or EDV results,and 436 had normal results for both.Patients with vascular ED had sig-nificantly lower estrogen levels(t=-3.546,P<0.001),lower testosterone levels(t=-2.089,P=0.037),and a higher rate of hyperglycemia(x2=12.772,P=0.002)compared with those with non-vascular ED.The patients with arterial ED were older(t=3.953,P<0.001),had a higher rate of hyperglycemia(x2=9.518,P=0.009),and a higher estrogen/testosterone ratio(t=2.330,P=0.020)compared with those with non-arterial ED.The patients with mixed arteriovenous ED had higher age(t=3.567,P<0.001),lower testosterone levels(t=-2.288,P=0.022),a higher rate of hyperglycemia(x2=12.877,P=0.002),and a larger estrogen/testosterone ratio(t=2.096,P=0.037)compared with those with normal findings.Multifactorial Logistic regression analysis indicated that higher levels of estrogen were a protective factor for vascular ED(OR=1.009,95%CI:1.004-1.014),and glucose 7.0 mmol/L was a risk factor(OR=0.381,95%CI:0.219-0.661).Older age was a risk factor for arte-rial ED(OR=0.960,95%CI:0.938-0.982).Additionally,older age(OR=0.976,95%CI:0.958-0.993)and glucose levels of 5.6-6.9 mmol/L(OR=0.591,95%CI:0.399-0.876)were also risk fac-tors for mixed arterio-venous ED.Conclusion:Hyperglycemia and aging may impair penile cavernous body vascular function,while higher levels of estrogen may have a protective effect on it.
		                        		
		                        		
		                        		
		                        	
6.Relationship between early urinary incontinence after holmium laser enucleation of the prostate and preoperative membranous urethral length
Ke LIU ; Fan ZHANG ; Ye YAN ; Chunlei XIAO ; Shudong ZHANG ; Yi HUANG ; Lulin MA
Chinese Journal of Urology 2023;44(1):21-25
		                        		
		                        			
		                        			Objective:To assess whether urinary incontinence after holmium laser enucleation of the prostate (HoLEP) is associated with membranous urethral length(MUL)on preoperative magnetic resonance imaging.Methods:The data of 96 patients who underwent HoLEP from January 2019 to April 2021 in Peking University Third Hospital were retrospectively analyzed. For all patients, the average age was (70.0±7.7) years old, the average body mass index was (23.9±2.9)kg/m 2, median pre-biopsy PSA was 3.79(2.48, 6.03)ng/ml, the average prostatic volume was (60.5±35.0)ml. 22 patients(22.9%) suffered with diabetes mellitus, and 17 patients(17.7%)had at least one time urinary retention. MUL was measured on MRI as the vertical distance from prostatic apex to the entry of the urethra into the penile bulb. All patients' median MUL was 13(11, 17)mm. The recovery of continence was followed up 2 weeks after HoLEP. The difference of age, body mass index, preoperative PSA, diabetes mellitus, urinary retention, prostate volume and MUL between urinary continence and incontinence group 2 weeks after HoLEP operation. The variables with P<0.1 were included in multivariable logistic regression to analyze the independent risk factors of urinary incontinence after HoLEP were compared. Results:All operations were successfully completed. The continence returned to normal in 72 cases (75.0%) and urinary incontinence existed in 24 cases (25.0%) in 2 weeks after surgery. There were 27 cases (37.5%) in continence group and 16 cases (66.7%) in incontinence group for those aged≥70 years. 21 cases (29.2%) in continence group and 13 cases (54.2%) in incontinence group had prostate volume ≥ 60 ml. There were 30 cases (41.7%) in continence group and 20 cases (83.3%) in incontinence group with MUL<13 mm. χ 2 test showed that age ( P=0.013), prostate volume ( P=0.027) and MUL ( P<0.001) were related to the incontinence after surgery. The age, prostate volume and MUL were included in the multivariate logistic regression analysis. Multiple logistic regression showed that MUL<13 mm( P<0.001) was independent predictor for incontinence after HoLEP. Conclusions:The incidence of urinary incontinence was high 2 weeks after HoLEP. Short MUL, which is less than 13 mm, is significantly associated with delayed recovery of urinary continence after HoLEP.
		                        		
		                        		
		                        		
		                        	
7.Influence of prostate cancer seminal vesicle invasion imaging classification on positive surgical margin after laparoscopic radical prostatectomy
Fan ZHANG ; Xinlong PEI ; Ye YAN ; Min LU ; Cheng LIU ; Shudong ZHANG ; Yi HUANG ; Lulin MA
Chinese Journal of Urology 2022;43(7):523-528
		                        		
		                        			
		                        			Objective:To investigate the effect of different imaging classifications of prostate cancer seminal vesicle invasion on positive surgical margins (PSM) after laparoscopic radical prostatectomy(LRP).Methods:114 patients with pT 3b stage prostate cancer admitted to Peking University Third Hospital from August 2009 to December 2020 were retrospectively analyzed. The age of the patients was (68.2±7.7) years old, the median pre-biopsy PSA was 20.20 (3.45-186.30) ng/ml, and the patients with biopsy Gleason score of ≤7, and ≥8 was 33 and 81 cases, respectively. The median prostate volume was 33.2 (12.1-155.4) ml. According to the imaging of the seminal vesicle invasion of prostate cancer, the patients were divided into the following types: type Ⅰ, the tumor directly invades the seminal vesicle along the vas deferens; type Ⅱa, the tumor invades the basal capsule of the prostate and invades the seminal vesicle; type Ⅱb, the tumor invades the periprostatic fat and retrogradely invades the seminal vesicles; type Ⅲ, solitary lesions in the seminal vesicles that do not continue with the prostate cancer. All patients underwent LRP, and the PSM were recorded as the base, bilateral, posterior, anterior and apical parts of the prostate. The differences in clinicopathological data of patients with different seminal vesicle invasion imaging types were compared, and the independent risk factors of PSM in pT 3b prostate cancer were evaluated by multivariate analysis. Results:The operative time of 114 cases in this group was (229.4±62.2) min, and the blood loss was 100(20-1 800)ml. The postoperative gross pathological Gleason score was ≤7 in 17 cases and ≥8 in 97 cases. In the imaging classification of prostate cancer with seminal vesicle invasion, there were 28 cases (24.6%) of type Ⅰ, 39 cases (34.2%) of type Ⅱa, 47 cases (41.2%) of type Ⅱb, and no type Ⅲ patients. There was no significant difference in age, body mass index, pre-biopsy PSA, prostate volume, and operation time among patients with type Ⅰ, Ⅱa, and Ⅱb seminal vesicle invasion ( P>0.05). There was a statistically significant difference in blood loss among the three types ( P = 0.001), and the difference in the proportion of lymph node metastasis was statistically significant ( P = 0.013). In the classification of prostate cancer seminal vesicle invasion, the PSM rates of type Ⅰ, Ⅱa and Ⅱb were 28.6% (8/28), 38.5% (15/39) and 70.2% (33/39), and the difference was statistically significant ( P=0.001). The PSM rates of type Ⅰ, Ⅱa, and Ⅱb were 21.4% (6/28), 23.1% (9/39), and 34.0% (16/47), respectively. The results of univariate analysis showed that the biopsy Gleason score ( P = 0.063) and the type of seminal vesicle invasion ( P<0.001) entered into multivariate analysis, and the results of multivariate logistic regression analysis showed that the type of seminal vesicle invasion ( P=0.001) was independent risk factor for PSM after LRP. Conclusions:The PSM rate in patients with type Ⅱb seminal vesicle invasion is significantly higher. The higher imaging type of seminal vesicle invasion is the independent risk factor of PSM after LRP.
		                        		
		                        		
		                        		
		                        	
8.Application of multidisciplinary teams online teaching in the teaching of urology residents based on network platform
Fan ZHANG ; Ye YAN ; Cheng LIU ; Hao WANG ; Min LU ; Wei HE ; Hongxian ZHANG ; Shudong ZHANG ; Yi HUANG ; Lulin MA
Chinese Journal of Medical Education Research 2022;21(2):215-218
		                        		
		                        			
		                        			Introducing the multidisciplinary cooperation model into the clinical teaching of residents has gradually been paid attention to, and the relevant multi-disciplinary teaching teams participate in and formulate teaching plan. The Department of Urology of the Peking University Third Hospital carries out multidisciplinary cooperative teaching of residents based on network platform to improve residents' autonomous learning ability and teaching effect. This model has certain advantages in mobilizing students' subjective initiative and cultivating learning interest. It is of great significance for the training of urology residents.
		                        		
		                        		
		                        		
		                        	
9.Relationship between positive surgical margin after laparoscopic radical prostatectomy and intravesical prostatic protrusion length
Fan ZHANG ; Yichang HAO ; Bin YANG ; Ye YAN ; Guoliang WANG ; Chunlei XIAO ; Shudong ZHANG ; Yi HUANG ; Lulin MA
Chinese Journal of Urology 2020;41(9):656-660
		                        		
		                        			
		                        			Objective:To identify the relationship between positive surgical margin after laparoscopic radical prostatectomy and intravesical prostatic protrusion length on preoperative magnetic resonance imaging.Methods:We retrospectively analyzed 110 patients with pathologic confirmed prostate carcinoma who underwent laparoscopic radical prostatectomy in our hospital. For all 110 patients, the average age was (70.4±7.8) years old, median pre-biopsy PSA was 12.23 ng/ml(range 0.78-110 ng/ml). There were 27 cases, 35 cases and 48 cases for biopsy Gleason score 6, 7 and ≥8. There were 73 patients with clinical stage T 1 and T 2, 37 patients with clinical stage T 3.The median prostatic volume was 38.16ml(range 11.83-163.36ml). MRI examination was performed in 1 week before the biopsy. Intravesical prostatic protrusion length (IPPL) was measured on MRI as the vertical distance from the tip of the protruding prostate to the base of the urinary bladder. All patients who underwent MRI preoperatively median IPPL was 3 mm(range 0-27 mm). There were 72 patients with IPPL<5 mm and 38 patients with IPPL≥5 mm respectively. All patients received extra-peritoneal laparoscopic radical prostatectomy. Parameters describing the surgical margin status and the location of positive surgical margin was recorded. The χ 2 tested the statistical significance in proportions differences. The multivariable logistic regression was used to assess risk factors for positive surgical margin and positive base surgical margin(PBSM). Results:Positive surgical margin rate was 38.1% for all patients, 25 patients(22.7%) had PBSM.χ 2 test showed that clinical stage ( P<0.001) and IPPL ( P=0.038) were related to the postoperative positive surgical margin. The clinical stage, Gleason score and IPPL were included in the multivariate logistic regression analysis. Multiple logistic regression showed that T 3 stage( P<0.001) was independent predictor for positive surgical margin. χ 2 test showed that clinical stage( P<0.001) and IPPL( P=0.001) were related to the postoperative PBSM. The clinical stage, Gleason score and IPPL were included in the multivariate logistic regression analysis. T 3 stage( P<0.001)and IPPL≥5 mm ( P=0.009) were independent predictors for PBSM according to multivariable logistic regression. Conclusions:For prostate cancer patients who received laparoscopic radical prostatectomy, clinical stage T 3was an independent risk factor for postoperative positive surgical margin. IPPL≥5 mm on preoperative magnetic resonance imaging and clinical stage T 3 were independent risk factors for PBSM.
		                        		
		                        		
		                        		
		                        	
10.Immunogenicity of the truncated NDV F protein surface-displayed on Lactobacillus casei.
Huanhuan LIU ; Shudong LI ; Yuqing YANG ; Xiaoying SUN ; Yan LI ; Xinyang LIU ; Xiaoyan CHEN ; Lianmei ZHANG ; Yongfei BAI ; Xilin HOU ; Liyun YU
Chinese Journal of Biotechnology 2019;35(8):1453-1462
		                        		
		                        			
		                        			To evaluate immune efficacy of the recombinant Lactobacillus casei, we constructed pLA-Newcastle disease virus (NDV)-F/L. casei and obtained the expression products. PCR amplified the NDV F gene carrying part of the major epitopes. The target gene was inserted to the shuttle plasmid pLA, and then transformed into Escherichia coli BL21 (DE3) in order to screen positive recombinant plasmid. The positive recombinant plasmid was transformed into L. casei by electroporation to construct pLA-NDV-F/L. casei. The positive strains were identified by PCR. The reactivity of the recombinant bacteria was identified by Western blotting and the protein expression was detected by indirect immunofluorescence, flow cytometry and laser confocal microscopy. The 14-day-old chickens in each group were vaccinated by oral plus nose drops. The pLA-NDV-F/L. casei twice immunization group and three times immunization group, the commercial vaccine group, the pLA/L. casei group, the unchallenge PBS and the challenge PBS group were established. IgG in serum and sIgA in the lavage fluid of intestinal, nasal and lung were detected by ELISA. The protection rate of chickens was evaluated. The results showed that 94.10% of the recombinant bacteria expressed the F protein. The recombinant protein was highly expressed on the surface of L. casei with a protein size of 62 kDa, which specifically bound to anti-NDV serum. The levels of anti-F IgG and sIgA antibodies in each test group were significantly higher than those in the control groups. The duration of antibody in the pLA-NDV-F/L. casei three-time immunization group lasted 28 days longer than that in the twice immunized group, and there was no significant difference between antibody peak values. The attack protection rates in each group of immunized pLA-NDV-F/L. casei three times, twice, attenuated vaccine, pLA/L. casei and PBS were 80%, 80%, 90%, 0% and 0%, respectively. Therefore, the antigenic protein of NDV F was successfully expressed by L. casei expression system, which has of reactogenicity and immunogenicity, and could induce protective immune responses in chickens.
		                        		
		                        		
		                        		
		                        			Animals
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		                        			Antibodies, Viral
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		                        			Chickens
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		                        			Immunization
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		                        			Lactobacillus casei
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		                        			Newcastle disease virus
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		                        			Vaccines, Attenuated
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		                        			Viral Vaccines
		                        			
		                        		
		                        	
            
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