Objective:To investigate the expression of programmed cell death ligand 1 (PD-L1), clinicopathologic features, immunohistochemical expression and molecular characteristics in fumarate hydratase (FH)-deficient renal cell carcinoma and to explore the potential application of immunotherapy in the patients.Methods:There were six patients with FH-deficient renal cell carcinoma treated at the First Affiliated Hospital of Fujian Medical University between January 2020 and October 2022. The clinical data, histological morphology, immunophenotype, PD-L1 expression and next-generation sequencing results were tabulated and analyzed.Results:There were 6 patients, all male, age ranged from 37 to 72 years (mean 45.7 years). Four cases were high-grade (WHO/ISUP grade3-4) with 2 or more histologic patterns, including papillary (most common), glandular, tubular, vesicular, ethmoid, nest-like, cystic and solid structures. Two cases were low-grade which showed nest-like, glandular, or tubular arrangement with eosinophilic flocculent cytoplasm and small intracellular vacuoles. Immunohistochemical analysis revealed strong expression of 2SC in all 6 cases, negative expression of FH in 5 cases, and positive expression of GATA3 in 5 cases. In high-grade cases, the mean values of CD4 and CD8 positive T-lymphocytes in advanced tumor invasion were 180.3/mm 2 and 130.5/mm 2, respectively. PD-L1 combined positive scores (CPS) were 20, 50, 5 and 30, respectively. The Ki-67 proliferative index were 20%, 20%, 10% and 30%, respectively. In low-grade cases, the mean values of CD4 and CD8 positive T-lymphocytes were 123.0/mm 2 and 100.5/mm 2, respectively. The PD-L1 CPS score was 1, and the Ki-67 proliferation index was 3%. High-throughput sequencing showed FH gene somatic mutation in 3 cases, FH gene germline mutation in 2 cases, and FH gene mutation was not detected in one case. Conclusion:FH-deficient renal cell carcinoma is more commonly high-grade than low grade. FH and 2SC are immunohistochemical markers used in the diagnosis of FH-deficient renal cell carcinoma, and GATA3 positivity is supportive of the diagnosis. The tumor infiltration of high-grade FH-deficient renal cell carcinoma shows an increase in CD4 and CD8 positive T-lymphocytes, and high expression of PD-L1; thus, anti-PD-L1 immunotherapy can be used as a treatment option.

To investigate the recent prevalence and molecular epidemiological characteristics of por-cine diarrhea viruses in Sichuan Province,this study used fluorescence quantitative PCR to detect porcine diarrhea samples from multiple regions in Sichuan Province from 2021 to 2023.RT-PCR was used to identify the genotypes of PEDV,PoRVA,PDCoV,and PTV,and their genetic variabil-ity,evolutionary characteristics,and recombination events were analyzed.The results showed that PEDV,PoRVA,PDCoV,and PTV were still prevalent in Sichuan region,with overall positive rates of 14.2％(40/281),13.2％(37/281),15.6％(44/281),and 12.5％(35/281),respectively.PEDV mixed infection with other pathogens was the most common.This study obtained a total of six strains of G2b PEDV,three strains of G3 PDCoV,three strains of G9P[13]PoRVA,one strain of G3P[13]PoRVA,three strains of Type 5 PTV,and one strain of Type 9 PTV.Compared to the seven vaccine strains including CV777,DR13,KPEDV-9,Chinju99,KNU-0801,AJ1102,and LW/L,the 6 PEDV strains showed multiple amino acid mutation sites in the COE region and S1D epitope region.Among them,the strains PSCLZ01 and PSCMY04 formed a separate branch in the phylogenetic tree.The three PDCoV strains have a closer genetic evolution distance to the previ-ously prevalent strains in Sichuan,but they also have 6-48 amino acid mutations compared to them.The four PoRVA strains have 104-108 amino acid variations in the VP4 gene compared to the early vaccine strain LLR,and they have 25 common amino acid variations in the VP7 gene.From the phylogenetic tree,the VP7 gene of RSCMY01/G3P[13]belongs to the same branch as the Heilongjiang strain LNCY,but its VP4 gene clusters with the Sichuan strain SCYA-C7,indica-ting that this PoRVA strain may have undergone genetic reassortment during inter-provincial transmission between different genotypes.It is worth noting that in the detected samples of PTV-5 and PTV-9,other diarrheal viruses tested negative,indicating that these two genotypes of PTV may be important pathogens causing porcine diarrhea.Additionally,the S gene of PEDV PSCLZ01 strain and PDCoV PCSCMY02 strain have undergone recombination events,and their parental strains come from different regions,both domestic and international.These findings reveal the main types of porcine diarrheal viruses,as well as their genetic diversity and variations in Sichuan Province in recent years.This study enriches the molecular epidemiological data of porcine diarrhe-al pathogens in the region and provides an important theoretical basis for the prevention,control,and purification of porcine diarrhea in the local area.

Objective:To investigate the clinicopathological features, immunophenotype, molecular characteristics and differential diagnosis of primary skull base chondrosarcoma.Methods:Nine cases of primary skull base chondrosarcoma were collected at the First Affiliated Hospital of Fujian Medical University, from January 2006 to June 2019, reviewed for the clinical and radiologic data and morphologic features, immunophenotype and molecular characteristics.Results:Among all the 9 cases, six were male, three were frmale, with average age 47 years, and median age 47 years; five cases were WHO gradeⅠ, and four were WHO grade Ⅱ. Microscopically, the tumor showed lobulated growth pattern with low-medium cellularity within a chondroid or mucoid background. The tumor cells showed mild-moderate atypia, with binucleated forms, and mitosis was rare or occasional. Immunohistochemistry (IHC) showed tumor cells were positive for S-100 protein, vimentin, SOX-9 and D2-40, and negative for Brachyury, CK, EMA and CK8/18; the Ki-67 index was low (1% to 5%). Molecular analysis showed IDH1 R132C mutation in four cases.Conclusions:Skull base chondrosarcoma is a rare cartilaginous malignant tumor with a good prognosis. Its characteristic morphologies, combined with IHC and molecular detection are helpful for the differential diagnosis.

Protein nitration and nitrosylation are essential post-translational modifications (PTMs) involved in many fundamental cellular processes. Recent studies have revealed that excessive levels of nitration and nitrosylation in some critical proteins are linked to numerous chronic diseases. Therefore, the identification of substrates that undergo such modifications in a site-specific manner is an important research topic in the community and will provide candidates for targeted therapy. In this study, we aimed to develop a computational tool for predicting nitration and nitrosylation sites in proteins. We first constructed four types of encoding features, including positional amino acid distributions, sequence contextual dependencies, physicochemical properties, and position-specific scoring features, to represent the modified residues. Based on these encoding features, we established a predictor called DeepNitro using deep learning methods for predicting protein nitration and nitrosylation. Using n-fold cross-validation, our evaluation shows great AUC values for DeepNitro, 0.65 for tyrosine nitration, 0.80 for tryptophan nitration, and 0.70 for cysteine nitrosylation, respectively, demonstrating the robustness and reliability of our tool. Also, when tested in the independent dataset, DeepNitro is substantially superior to other similar tools with a 7%-42% improvement in the prediction performance. Taken together, the application of deep learning method and novel encoding schemes, especially the position-specific scoring feature, greatly improves the accuracy of nitration and nitrosylation site prediction and may facilitate the prediction of other PTM sites. DeepNitro is implemented in JAVA and PHP and is freely available for academic research at http://deepnitro.renlab.org.
Amino Acid Sequence ; Amino Acids ; metabolism ; Deep Learning ; Humans ; Internet ; Neural Networks (Computer) ; Nitrosation ; Proteins ; chemistry ; metabolism ; Reproducibility of Results ; Software

Objective To observe the changes of proliferation and apoptosis of colon cancer cell line treated with oxaliplatin after the downregulation of ERRα and to investigate the mechanism.Methods Colon cancer cell lines Colo-205,HCT-116,SW620 and HT-29 were cultured by adherent cells and in accordance with the given intervention,they were divided into group XCT790-OHP-HCT-116(after oxaliplatin treatment,ERR αinhibitor XCT790 was administered),group siERRα-OHP-HCT-116(after oxaliplatin treatment,siERR α was transfected into HCT-116 cells and downregulated ERR αexpression),oxaliplatin intervention group(group OHP-HCT-116)and the control group(NC group)which was given no intervention.The experiment was divided into siERR αgroup with siERR α transfected with HCT-116 cells,downregulated ERR αexpression and the negative control group(siNC group)transfected with siNegative Control.Using Western blot method and real-time quantitative(qRT)-PCR for the detection of colorectal cancer cell ERRαprotein and mRNA expression,the expression of ERR αwas downregulated by ERR αinhibitors XCT790 and siERR,and apoptosis and proliferation of colon cancer cells were detected by flow cytometry and MTT.Western blot and qRT-PCR were used to detect apoptosis and proliferation-related gene proteins and mRNA expression.Results ERR αand mRNA protein in HCT-116 were higher than those of Colo-205,SW620 and HT-29 cell lines(P<0.05); in the XCT790-OHP-HCT-116 group,the early apoptosis rate was higher than those of the NC group and OHP-HCT-116 group(P<0.05),the survival rate of cell culture in 72 and 96 h in the XCT790-OHP-HCT-116 group was lower than those in the NC group and OHP-HCT-116 group(P<0.05).The siERR α HCT-116 cells transfected with down-regulation of ERR expression,siERR α -OHP-HCT-116 group early apoptosis rate was lower than those of NC group and OHP-HCT-116 group(P<0.05),siERR -OHP-HCT-116 group cells cultured for 72 and 96 h after the survival rate was lower than the NC group and OHP-HCT-116 group(P<0.05);After the downregulation of ERRαby siERR alpha transfected with HCT-116 cells,the early apoptotic rate in the group siERRα-OHP-HCT-116 was lower than that in the group NC and group OHP-HCT-116(P<0.05),the survival rate of the group siERRα-OHP-HCT-116 after 72 and 96 h were lower than those in the group NC and group OHP-HCT-116(P<0.05),siERR α was transfected into HCT-116 cells,compared with the siNC group,YAP1,p73,p63,MDM2, Capase 8,Capase 9 protein in the siERR group decreased(P<0.01),there was no significant difference in the level of mRNA(P>0.05).Conclusion The downregulation the expression of ERRαcan promote colon cancer cell apoptosis,inhibit proliferation,and enhance the killing effect of oxaliplatin on colon cancer cells.

ObjectiveTo investigate the efficacy and toxic and adverse effects of high-dose hypofractionated radiotherapy in elderly patients with stage Ⅳ pancreatic cancer. MethodsThe clinical data of the patients with pancreatic cancer and distant metastasis who were admitted to our hospital from September 2011 to May 2015 were collected, and all the patients underwent high-dose hypofractionated helical tomotherapy. The data on efficacy and toxic and adverse effects were obtained through follow-up, and the evaluation of adverse effects was performed according to National Cancer Institute-Common Terminology Criteria for Adverse Events version 4.02. The Kaplan-Meier method was used for survival analysis. ResultsA total of 33 patients older than 65 years received the high-dose hypofractionated radiotherapy. Of all the patients, 30 received follow-up visits, and the follow-up rate was 91.0%. The median survival time was 9 months, the 1-year overall survival rate was 24.0%, and the rate of pain relief was 80.0% (20/25). The treatment outcome of pancreatic lesions could be evaluated in 17 patients, among whom 4 (23.5%) achieved partial remission, 12 (70.6%) achieved stable disease, and 1 (5.9%) experienced progression. As for toxic and adverse effects, the incidence rate of grade 3 hematologic toxicity was 6.7% (2/30), and no patients experienced grade ＞2 upper gastrointestinal reactions. ConclusionIn elderly patients with stage IV pancreatic cancer, high-dose hypofractionated radiotherapy has tolerable toxic and adverse effects and can relieve cancer pain and prolong survival time.

Objective:To explore the relationship of CDH17 expression with clinico-pathological features and the correlation be-tween the single nucleotide polymorphisms (SNPs) of CDH17 gene and genetic susceptibility of gastric carcinoma (GC). Methods:A tissue microarray was performed to simulate the dynamic process of invasion and metastasis of GC. Immunohistochemical staining was performed to detect the expression of CDH17 protein, and PCR-based LDR was performed to detect the 2 SNP loci (rs2514813 and rs3214050) genotypes of CDH17 gene. Results: The expression of CDH17 protein in GC was more significantly up-regulated and greatly increased in the intestinal type than in the diffuse type. The expression of CDH17 protein in GC was positively correlated with the histological grading (P<0.01) and was not associated with the survival (P=0.209). With the progression of the cancer invasion, the expression of CDH17 protein in GC showed a downtrend from the gastric mucosa layer to the invasive front edge. The frequencies of the C and T alleles and the CC, CT, and TT genotypes at the CDH17 rs3214050 locus between the GC patients and the control groups were significantly different (P<0.01). However, no significant differences were observed at rs2514813 (P>0.05). The individuals with the T al-leles had longer survival time than those with the CC genotype (P<0.01). Conclusion:The up-regulation of CDH17 expression is in-volved in the maintenance of histological phenotype and progression of GC. Individuals with T alleles at the CDH17 rs3214050 locus have decreased risk of GC and had better prognosis (OR=0.762, 95%CI:0.619-0.937), thereby suggesting that screening for these alleles would help with the assessment of genetic susceptibility and prognosis of GC in the Fujian population.

Background and purpose: PDCD4 may be inhibited by miR-21 to regulate the malignant behaviors of colon cancer such as invasion and migration. This study aimed to explore the function of colon cancer HT-29 cell lines by downregulating miR-21 expression and discuss the mechanisms and relationship between miR-21 and PDCD4 in colon cancer malignant behaviors. Methods:simiR-21 was transfected into colon cancer cell line HT-29 to downregulate the expression of miR-21. Proliferation, apoptosis, migration and invasion were detected by MTT, flow cytometry and Transwell assay after transfection. PDCD4 expression was detected by Western blot and qRT-PCR. Results:The qRT-PCR analysis result proved that the transfection efifciency was 60%-65%. MTT analysis result showed that the proliferations of HT-29 cells were inhibited after the transfection of miR-21 for 72, 96, 120 h (t=1.276, P<0.05;t=3.276, P<0.01;t=4.523, P<0.01). Comparing with si-negative control and miR-21 groups, lfow cytometry result showed that the apoptosis rate was increased after miR-21 expression downregulated (t=2.132, P<0.05;t=3.524, P<0.05). Transwell assay result showed that migration (t=2.423, P<0.05; t=3.153, P<0.05) and invasion(t=3.245, P<0.05; t=5.236, P<0.05) were inhibited;Western blot result showed that PDCD4 expression was up-regulated at protein level(t=2.342, P<0.05;t=4.215, P<0.05);qRT-PCR result showed that PDCD4 expression was up-regulated at mRNA level(t=2.261, P<0.05; t=3.492, P<0.05). Conclusion: The proliferation, migration and invasion are the inhibited, and apoptosis is attenuated after miR-21 downregulated by simiR-21 transfection, PDCD4 expression is up-regulated. miR-21 may enhance the malignant behavior of cancer cells by downregulating the PDCD4 expression, miR-21 might be a target gene for colon cancer therapy.

Objective:To investigate the correlation of Mena protein expression with the invasion and metastasis of Mena SNPs with genetic susceptibility in gastric cancers (GC). Methods:A tissue microarray that simulates the invasion and metastasis process of GC was created, and immunohistochemistry was performed to detect the expression of Mena protein. The Mena gene 5 SNP loci geno-types of 188 healthy people and 389 GC patients were assayed using PCR-based LDR analysis. Results:The expression of Mena pro-tein in GC was significantly upregulated and greatly increased in the intestinal-type and mixed-type GC than that in the diffuse-type and was negatively related to the invasion and metastasis of GC. Patients with Mena overexpression had better prognosis. The frequen-cies of the A and G alleles, as well as the AA, AG, and GG genotypes, at the Mena SNP rs3795443 locus were significantly different be-tween patients with gastric carcinoma and the control groups (OR=2.1489,95%CI 1.4607~3.1613, P<0.01). The frequencies of these five Mena gene SNP loci were not significantly related with the survival of patients with gastric carcinoma. Conclusion:The upregula-tion of Mena expression is involved in maintaining the histological phenotype, invasion, metastasis, and prognosis of gastric adenocarci-noma. Individuals with GG and AG genotypes at the Mena rs3795443 locus have increased risk of gastric carcinoma, which suggests that screening for this genotype would be helpful in assessing the genetic susceptibility of gastric carcinoma.

The objective of this study is to compare the normalized prediction distribution errors (NPDE) and the visual predictive check (VPC) on model evaluation under different study designs. In this study, simulation method was utilized to investigate the capability of NPDE and VPC to evaluate the models. Data from the false models were generated by biased parameter typical value or inaccurate parameter inter-individual variability after single or multiple doses with the same sampling time or multiple doses with varied sampling time, respectively. The results showed that there was no clear statistic test for VPC and it was difficult to make sense of VPC under the multiple doses with varied sampling time. However, there were corresponding statistic tests for NPDE and the factor of study design did not affect NPDE significantly. It suggested that the clinical data and model which VPC was not fit for could be evaluated by NPDE.