Objective To explore the effects of specific T-cell immunity against prostate cancer(PC) cells induced by dendritic cells(DCs) derived from fetal organs.Methods Mononuclear cells(MNCs) were obtained from the fetal bone marrow and liver.Then MNCs were cultured in medium with induction of rhGM-CSF,rhIL-4 and rhTNF-?to get DCs.Lysates of DU145 containing HSP-peptide complex were prepared by 50%-70%(NH4)2SO4 saturation.T lymphocytes from fetal spleen were co-cultured with DC loading DU145 antigen for 72 h,whereby CTL was obtained.The cytotoxicity of CTL against DU145,PC3 and EJ was detected by MTT assay.Results Mature DCs were induced from fetal organs,which expressed CD1a,CD_(86),HLA-DR and CD_(83) at high levels.DC stimulated with tumor lysates transformed T cells to specific CD~+_8 CTL.Phenotype of CD~+_8 cell was(14.09?(2.46))% before transformation,and(62.76?2.64)% after transformation,respectively(P

Objective To investigate the clinical and pathological characteristics of nephrogenic adenoma. Methods Eleven patients were diagnosed as nephrogenic adenoma including 5 men and 6 women, aged 37-78 years (56 on average). The pathological findings in all cases of nephrogenic adenoma were presented with a review of the literature. Results Eleven cases of nephrogenic adenomas were evaluated, 2 cases were in ureter and 9 cases were in the bladder. Eight of the 9 bladder cases underwent TUR-BT surgery in continuous epidural anesthesia, 1 case underwent partial cystectomy with general anesthesia. A right ureteroscopy and left ureterolithotomy were performed respectively in continuous epidural anesthesia for the 2 cases in ureter. The final diagnosis was based on histopathological findings. For all of cases, 8 cases were diagnosed as nephrogenic adenomas, 2 cases as atypical nephrogenic adenoma and 1 case as nephrogenic adenoma with malignant transformation. The microscopic appearance of nephrogenic adenoma demonstrated that morphology closely resembled aberrant tubules of the kidney. In addition, atypical nephrogenic adenomas appeared as the presence of cytologic atypia, including nuclear enlargement, nuclear hyperchromasia and prominent nucleoli. The morphologic changes of nephrogenic adenomas with malignant transformation were that tumor cells retained the basic structural characteristics of typical nephrogenic adenomas, and the similar morphological cells lost adhesion ability among cells and presented diffuse solid growth in the surrounding area.Intravesical perfusion was further performed for treating the patients with atypical nephrogenic adenomas or nephrogenic adenomas with malignant transformation. The mean patient follow up was 46 months (range, 24- 104 months), and there was only 1 case of recurrence. Conclusions Nephrogenic adenoma is an uncommon benign lesion of the urinary tract. The symptoms and cystoscopic manifestations are not unique. We reported one patient of nephrogenic adenomas with malignant transformation and provided some evidence for malignant alteration in morphology and invasive behavior. All patients underwent local excision of the lesions. Intravesical perfusion was further performed for treating the patients of atypical nephrogenic adenomas or nephrogenic adenomas with malignant transformation. Whether it is nephrogenic adenoma or atypical nephrogenic adenoma, long-term follow-up after treatment is necessary.

Objective To investigate the histopathologic characteristics of bladder tumor and provide theoretical basis for the reasonable selection of treatment modality.Methods This retrospective study collected the pathological data of 4 200 bladder tumor from May 2001 to October 2014.There were 3 443 male and 757 female, and the average diameter of these tumors was (1.8 ± 0.6) cm (ranged 0.2 to 6.5 cm).Among all cases, 3 214 (76.5％) cases were solitary tumor while 986 (23.5％) were multiple tumors.The histologic subtype, pathological grade and stage, the existence of vascular and lymphovascular invasion, tumor in situ, abnormal variants and rare subtypes were recorded and analyzed.Results 162 cases (3.9％)were benign tumors and 4 038 cases (96.1％)were malignant tumors including 4 008 cases of urothelial cancer (UC), 18 cases of primary adenocarcinoma and 12 cases of primary bladder squamous carcinoma.Furthermore, 2 460 (61.4％)cases were high grade UC while 1 548(38.6％)cases were low grade.320 cases were found intravascular tumor embolus or lymphovascular tumor thrombus and 391 (9.3％)cases were found metaplasia of squamous epithelium.Moreover, there were 230 cases of squamous differentiation, 120 cases of glandular differentiation, 110 cases of both squamous and glandular differentiation, and 39 cases (0.9％)of other rare subtypes or variations.On pathological stage, 112 (2.8 ％) cases were carcinoma in situ, 548 (13.7％)cases were Ta, 2 599(65.1％)cases were T1, 480(12％)cases were T2, 92 cases(2.3％)were T3 and 23 cases(0.6％)were T4 stage, with the rest cases being unable to be accurate staging.Multiple Logistic regression analysis revealed that lymphovascular invasion was related to tumor grade , pathological stage and abnormal differentiation (P ＜ 0.02).Moreover, UC with squamous and glandular differentiation were related with tumor recurrence and progression (P =0.02).Conclusions Most bladder tumors were high grade and low stage urothelial cancer with various forms of differentiation.Squamous and glandular differentiation were most common variation which should be avoided to diagnosed as hybrid carcinoma.Lymphovascular tumor thrombus and abnormal differentiation were correlated with tumor stage and grade.

Objective To study the clinical and histopathologic features of small renal carcinoma (diameter≤4 cm)and provide theoretical basis for evaluating the safety,efficacy and prognosis of nephron sparing surgery.Methods This retrospective study collected the pathological data of 490 patients with small renal cell carcinoma,who were treated in our hospital,from May 2000 to October 2014.We recorded and analyzed the tumor size,histological subtype,Fuhrman grading,pathological stage,the existence of mulifocality,vascular invasion,tumor psuedocapsule,hemorrhage or necrosis and distant metastasis.Results The median diameter of tumor was (3.2 ± 0.6) cm,ranged 0.6 to 4.0 cm.Of all the subjects,422 (86.1％) were clear cell carcinoma,32 (6.5％) were chromophobe cell carcinoma,23 (4.7％) were papillary carcinoma and 13 (2.7％) were other rare types.Among the 422 clear cell carcinoma cases,27 were Fuhrman grade Ⅰ,157 were Ⅰ-Ⅱ grade,210 were grade Ⅱ,21 were Ⅱ-Ⅲ grade,7 were grade Ⅲ and no one was grade Ⅳ.Multifocal tumors were found in 18 cases (3.7％) and tumor embolus of renal vein was found in 6 cases (1.2％).Intact psuedocapsule were found in 326 (66.5％) tumors with the thickness ranged from 0.2 to 1.0 mm.Tumor infiltration without the psuedocapsule penetration were found in 82 cases (16.7％),penetrated into the psuedocapsule were found in 11 cases (2.2％),infringement of perirenal fat were found in 9 cases (1.8％).Hemorrhage and necrosis were found in 240 cases (48.9％),synchronous lung metastases occurred in 3 patients (0.6％).Logistic regression analysis revealed that tumor invasion and pseudocapsule penetration were related to Fuhrman Ⅱ-Ⅲ,Ⅲ and tumor diameter (P =0.04).Moreover,tumor size was related with histological grade and renal capsule invasion (P =0.02).Nevertheless,there was no relationship among tumor size,renal vein embolus or mulifocality (P =0.35).Conclusions Although most small renal tumors are high differentiation and low grade,but rare cases are aggressive with infringement of perirenal fat or early distant metastasis,suggesting heterogeneity in its biological behavior.Most small renal tumors have obvious psuedocapsule.When the tumor size is greater than 3.0 cm and its Fuhrman classification was high,the psuedocapsule and perirenal fat are more likely to be infiltrated.Nephron sparing surgery should remove the tumor and its surface adipose tissue entirely.

Objective To study the gene mutation of fibroblast growth factor receptor 3 (FGFR3) and p53 in bladder cancer tissue and to explore their relationship with tumor recurrence. Methods DHPLC and PCR direct sequence were used to detect the mutation of FGFR3 and p53 in BTCC (n=98) and normal bladder mucosa (n=10). Genomic DNA of 98 BTCC was extracted. The exon 5-8 of P53 and the exon 7, 10, 15 were amplification by PCR. The products of PCR was screened by DHPLC to detect the mutation of the production. The results of the FGFR3 and p53 mutation were analyzed by Kaplan-Meier method and no recurrence survival rate was tested by log rank test. All the analysis were aim to explore the clinical biological value of the mutation of FGFR3 and p53. Results Mutation of FGFR3 in BTCC (44. 9%) was higher than normal bladder mucosa(0, P<0.01). Mutation in T_a-T_1 was 75. 6%(33/45) ,T_2 -T_4 was 26. 6%C10/53). Mutation in G_1 was84. 6%(11/13),inG_2 was 61. 4% (27/44), in G_3 was 14. 6% (6/41), (P<0. 05). The mutation rate was lower with the higher of stage and grade. Mutation of p53 in BTCC (34. 6%) was higher than normal bladder mucosa (0%) (P<0. 01). Mutation in T_a - T_1 was 20. 0% (9/45), T_2 - T_4 was 47. 2%(25/53). Mutation in G_1 was G_1 7. 7%(1/13), in G_2 18. 2%(8/44),in G_3 58. 1%(25/41) , (P<0. 05). The mutation rate was higher in the higher stage and grade. Kaplan-Meier method results revealed that mutation of FGFR3 indicating a favorable prognosis while mutation of p53 indicating a poor prognosis. As to the analysis of genotype, the type of FGFR3mut/p53wt had a relative longer recurrent interval (P<0. 01). Conclusions Mutation of FGFR3 indicated a relative longer recurrent interval, which revealed a favorable prognosis of BTCC. Mutation of p53 indicated a relative shorter recurrent interval, which revealed a poor prognosis.

Objectives:To investigate the prognostic value of soluble growth stimulation expressed gene 2 protein(sST2)combined with N-terminal pro-brain natriuretic peptide(NT-proBNP)in patients with ST-segment elevation myocardial infarction(STEMI)undergoing primary percutaneous coronary intervention(PCI). Methods:A total of 206 patients who were diagnosed with STEMI for the first time and underwent emergency PCI from August 2020 to February 2021 in the People's Hospital of Liaoning Province were enrolled.Patients were followed up for 3 years and divided into major adverse cardiac event(MACE,a composite endpoint event including cardiac death,stroke,heart failure,and ischemia-driven revascularization)group and MACE-free group.Multivariate cox analysis was performed to determine the independent risk factors for the prognosis of primary PCI in STEMI patients;the predictive value of sST2 and NT-proBNP for the occurrence of MACE in STEMI patients undergoing primary PCI was assessed by ROC analysis and the prediction of MACE by each factor by itself and the combined variables was analyzed with the Delong test. Results:There were 62 cases of MACE during the 3-year follow-up.Compared with the MACE-free group,patients in the MACE group had higher levels of sST2 and NT-proBNP,higher proportion of patients with left anterior descending branch lesions,anterior wall myocardial infarction,lower LVEF,and higher proportion of coronary artery slow flow(all P<0.05).Multivariate Cox analysis showed that sST2(HR=1.018,95%CI:1.012-1.024,P<0.001)and NT-proBNP(HR=1.001,95%CI:1.000-1.010,P<0.001)were independent predictors of MACE.According to the statistical analysis of ROC and Delong test,the AUC of combined sST2 and NT-proBNP in predicting MACE was 0.854,the sensitivity was 64.5%,the specificity was 93.1%,and the combined prediction of prognosis was better than that of individual prediction,with statistically significant difference(Z=2.119,P=0.034;Z=2.178,P=0.029). Conclusions:Serum sST2 and NT-proBNP are valuable predictors of MACE after emergency PCI in patients with STEMI,and the predictive efficacy increases with combined assessment of both sST2 and NT-proBNP.

Objective To summarize the clinical and electroencephalogram features of neuronal ceroid lipofus-cinosis (NCL). Methods A retrospective analysis of the clinical phenotypes and electroencephalogram features of pa-tients diagnosed with NCL in Department of Pediatrics,Peking University First Hospital from February 2000 to August 2015 were conducted. Results Among the 30 patients,18 were male and 12 were female. The age of onset was between 9 months to 7 years old. The first symptoms included seizure in 22 patients,psychomotor developmental delay or regre-ssion in 7 cases,and visual loss in 1 case. Clinical manifestations included psychomotor regression in 29 cases,epilepsy in 28 cases,visual impairment in 19 cases,ataxia in 20 patients,and positive pyramidal tract sign in 13 cases. Twenty-one patients accepted fundus oculi examination. Seven patients were found with macular degeneration,8 cases with optic nerve atrophy,2 cases with retinal pigment degeneration,and 8 patients were normal. Brain atrophy were found in all 30 cases,including diffuse brain atrophy in 14 cases,only cerebellar atrophy in 6 cases,and cerebral atrophy with periven-tricular T2W high signal in 10 cases. Video electroencephalogram(EEG)examination was performed in 27 patients and their backgrounds were diffuse slow waves. Seven patients didn't have physiological vertex sharp waves or sleep spin-dles. Generalized epileptiform discharges were captured in 6 cases,focal epileptiform discharges in 15 cases. Both of generalized and focal epileptiform discharges were captured in 6 cases. Generalized slow wave burst in 4 cases,and in-termittent photic stimulation evoked epileptiform discharges in 3 cases. Ten patients were observed with clinical sei-zures,including 4 cases of myoclonic episodes,3 cases of atypical absences,3 cases of focal seizures,1 case of atonic and one of tonic spasms. Peripheral blood enzyme examination was taken in 13 patients,among whom 8 patients were identified with tripeptidyl peptidase 1 (TPP1)deficiency and 1 patient with palmitoyl protein thioesterase 1 (PPT1) deficiency. Twenty-eight patients accepted skin and/or muscle electron microscope examination. Osmiophilic granular was found in 2 cases,curvilinear bodies in 15 cases,fingerprint profiles in 2 cases,curvilinear and linear bodies in 1 case,fingerprint profiles and osmiophilic granular in 1 case. NCL-related gene detection was conducted in 3 patients, with 1 patient identified with CLN6 compound heterozygous mutations and 2 patients with TPP1 homozygous mutations. Thirty patients were classified into 3 groups based on the onset age,enzymatic examination results and pathological examination of skin and muscle,including 5 cases of infantile NCL,20 cases of late-infantile NCL,and 5 cases of juvenile NCL. Conclusions The clinical features of NCL included multiple types of epileptic seizures (among which myoclonus was the most common type),psychomotor developmental delay or regression,vision loss,ataxia,and positive pyramidal tract sign. Its MRI was characterized with brain atrophy. EEG showed diffuse slow wave activity,with focal and/or generalized epileptiform discharges. Specific enzyme examination,and skin or muscle pathology or gene test could help to make diagnosis.

This paper comprehensively discusses on the potential neurobiological mechanisms of acupuncture in the treatment of tobacco dependence， focusing on three important aspects， including acupuncture's regulation of tobacco dependence behavior， effects of acupuncture on withdrawal syndrome， and the role of acupuncture in preventing relapse. It is found that acupuncture can inhibit drug-seeking behavior by regulating the reward pathway and related neurons， such as dopamine， thus modulating tobacco dependence behavior. It also alleviates withdrawal symptoms by improving the oral environment of smokers and reducing negative emotions after quitting. Furthermore， acupuncture can prevent relapse by decreasing brain network activity related to smoking cravings and improving cognitive brain functions like addiction memory. Currently， research on the specific neurobiological mechanism of acupuncture in treating tobacco dependence and the involved neural circuits is limited. Future research directions are proposed， including the evaluation of clinical effects， exploration of specific therapeutic mechanisms， investigation of brain pathology， and strengthening the exploration of brain functions. Additionally， combining modern technologies to clarify the neural circuits involved in acupuncture intervention will provide a basis for acupuncture treatment of tobacco addiction.

Objective To investigate the expression of Midkine(MDK)in cholangiocarcinoma(CCA)and its value in predicting the prognosis of CCA,as well as the potential mechanism of the effect of MDK on the progression of CCA.Methods The data of CCA samples were obtained from TCGA database to analyze the difference in the expression of MDK between cancer tissue and paracancerous tissue and its association with clinical features,and the data collected from GEO database and 11 CCA patients who underwent surgical resection in The Fifth Medical Center of Chinese PLA General Hospital from June 2018 to September 2021 were used for validation.STRING and Cytoscape were used to construct a protein-protein interaction network,and gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses were used to investigate the biological functions and tumor-related pathways involving MDK-related genes.In addition,TIMER and TISIDB databases were used to analyze the correlation between MDK expression and immune cell infiltration in CCA tissue.The independent-samples t test or the Mann-Whitney U test was used for comparison of continuous data between two groups,and the Fisher's exact test was used for comparison of categorical data between two groups.The Kaplan-Meier method was used to plot survival curves,and the Log-rank test was used for comparison between groups.The Spearman correlation analysis was used to investigate the correlation between two variables.Results The expression level of MDK in cancer tissue and paracancerous tissue of CCA patients was compared based on TCGA database,and the results of the non-paired and paired analyses showed that the expression level of MDK in CCA tumor tissue was significantly higher than that in paracancerous tissue(P<0.001).Transcriptome sequencing was performed for the tumor tissue and its corresponding paracancerous tissue from 11 CCA patients,and the results showed that the expression level of MDK in CCA tumor tissue was significantly higher than that in corresponding paracancerous tissue(P<0.01).High expression of MDK was associated with lymph node metastasis(P=0.045)and vascular invasion(P=0.044).Survival analysis showed that compared with the CCA patients with low MDK expression,the CCA patients with high MDK expression had significantly shorter overall survival time(χ2=5.30,P=0.028)and disease-specific survival time(χ2=6.25,P=0.019).The GO and KEGG enrichment analyses showed that the 30 MDK-related genes were closely associated with ubiquitin-mediated proteolysis and affected the prognosis of CCA patients.The TIMER analysis showed that the expression level of MDK was positively correlated with the infiltration of B cells(r=0.356,P=0.035 6)and dendritic cells(r=0.409,P=0.014 7)in tumor microenvironment of CCA;the TISIDB analysis showed that the expression level of MDK was positively correlated with CXCL16(r=0.465,P=0.004 67)and was negatively correlated with CXCL12(r=-0.389,P=0.019 7)and CXCR5(r=-0.393,P=0.018 5),and it was also negatively correlated with the immune checkpoint regulators VTCN1(r=-0.393,P=0.018 3),LTA(r=-0.380,P=0.022 7),and PVR(r=-0.350,P=0.037 3).Conclusion High expression of MDK is associated with poor prognosis in CCA patients,and MDK has the potential of being used as a molecular marker for predicting the prognosis of CCA.MDK may promote the development and progression of CCA by regulating ubiquitin-mediated proteolysis and the infiltration of B cells and dendritic cells.

ObjectiveSerum metabolomics of acute lung injury（ALI） in rats was conducted using ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS） to explore the similarities and differences in the mechanism of Qingqiao（harvested when the fruits of Forsythiae Fructus were initially ripe and still green in color） and Laoqiao（harvested when the fruits of Forsythiae Fructus were ripe） in the treatment of ALI. MethodA total of 24 SD male rats were acclimatized and fed for 1 week， 6 of them were randomly selected for the blank group and 18 for the experimental group. The ALI model was induced in the experimental group by tracheal intubation with lipopolysaccharide（LPS）. After successfully constructing the ALI model， these rats was randomly divided into model group， Qingqiao group and Laoqiao group， with 6 rats in each group. The Qingqiao and Laoqiao groups were administered orally once a day at a dose of 1.5 g·kg^-1， while the blank and model groups received an equivalent volume of saline for 3 consecutive days. The pathological conditions of rat lung tissues were comprehensively assessed by hematoxylin-eosin（HE） staining， wet-to-dry mass ratio（W/D） of lung tissues， and protein concentration in rat bronchoalveolar lavage fluid（BALF）. The levels of interleukin（IL）-6， IL-1β and tumor necrosis factor（TNF）-α in BALF were quantified using enzyme-linked immunosorbent assay（ELISA）. UPLC-Q-TOF-MS was used to identify and analyze the chemical compositions of Qingqiao and Laoqiao， and serum metabolomics of rats in each group was analyzed， combined with multivariate statistical analysis with variable importance in the projection（VIP） value>1， P<0.05 from t-test， and fold change（FC）≥1.5 or FC≤0.5 to screen the differential metabolites Qingqiao and Laoqiao for the treatment of ALI. The Kyoto Encyclopedia of Genes and Genomes（KEGG） database was used in combination with MetaboAnalyst for the metabolic pathway analysis of the screened differential metabolites. ResultCompared with the blank group， rats in the model group exhibited enlarged alveolar lumen， ruptured alveoli， interstitial hemorrhage， bronchial exudation of a large number of neutrophils and erythrocytes， and a significant increase in the protein concentration in the BALF and the W/D value of the lung tissues（P<0.01）. In contrast， compared with the model group， rats in the Qingqiao group and the Laoqiao group showed reduced bronchial hemorrhage in the lungs， and the protein concentration in the BALF and the W/D value of the lung tissues were significantly decreased（P<0.01）， the lung injury was significantly alleviated， but more obvious in the Qingqiao group. Compared with the blank group， the expression levels of IL-6， IL-1β and TNF-α in the BALF of the model group were significantly higher（P<0.01）. Additionally， compared with the model group， the expression levels of IL-6， IL-1β and TNF-α in the Qingqiao and Laoqiao groups were significantly lower（P<0.01）. The chemical composition analysis of Qingqiao and Laoqiao revealed that 63 components were detected in Qingqiao and 55 components were detected in Laoqiao， with 47 common components， 16 components unique to Qingqiao and 8 components unique to Laoqiao. Characterizing the differences in serum metabolomics in rats， 19 and 12 metabolites were called back by Qingqiao and Laoqiao， respectively. The metabolic pathway enrichment analysis showed that Qingqiao exerted its therapeutic effects by affecting 6 key metabolic pathways， including linoleic acid metabolism， phenylalanine metabolism， phenylalanine， tyrosine and tryptophan biosynthesis， glycerophospholipid metabolism， α-linolenic acid metabolism， and arachidonic acid metabolism， and Laoqiao exerted therapeutic effects by affecting 6 key metabolic pathways， including linoleic acid metabolism， arachidonic acid metabolism， sphingolipid metabolism， phenylalanine metabolism， ascorbate and aldarate metabolism， and glycerophospholipid metabolism. ConclusionQingqiao and Laoqiao have therapeutic effects on ALI， and Qingqiao is more effective. Both of them can play a therapeutic role in ALI by regulating amino acid metabolism and lipid metabolism， but the metabolic pathways affected by them are different.