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.

Capillary zone electrophoresis (CZE) was developed to investigate the structure stability of novel camptothecin derivative (L-P) at different pH,the kinetics and thermodynamics of hydrolysis reaction from lactone form to carboxylate form direction at near physiological conditions (pH 7.4,310 K). Uncoated fused-silica capillaries(35 cm×50 μm i. d,with effective length of 26.5 cm) were used. The background electro-lyte( BGE) was 0.025 mol/L sodium phosphate buffer with pH varied at 2.5,4.0,5.0,6.0,7.0,7.4 and 9. 0. The electrophoresis voltage was maintained at 14 kV when the pH of BGE ranged between 2.5 and 5.0,otherwise,the voltage was maintained at 10 kV. The UV detector was set at 260 nm. All samples were introduced using hydrodynamic injection at 5 kPa for 4 s. L-P was found to be lactone form as the solution pH was below 4. 0. As pH increased,the lactone form of L-P would undergo hydrolysis reaction to be carboxylate form. As pH was 9.0,L-P existed almost completely as carboxylate form. The rate constant of the hydrolysis increased as temperature raise. The energy of activation ( Ea) ,the enthalpy ( ΔH) and entropy (ΔS) of the hydrolysis reaction were determined as 72. 6 kJ/mol,10. 5 kJ/mol and 50. 9 J/( mol K) ,respectively. The proposed capillary zone electrophoresis could efficiently separate two pH-dependent structural forms of the novel camptothecin derivative( L-P). The positive enthalpy and entropy values of the L-P hydrolysis indicated that the reaction was endothermic and entropically driven and higher temperature favored.

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.

When Playmonas cardiformis is exposed to different ZnSO4 concentration, its growth rate decreases gradually with the increase of ZnSO4 concentration which is more than 1 mmol?L~-1. More than 8 mmol?L~-1 ZnSO4 can lead to the death of playmonas subcordi- formis. But the production of MT-like increases with the increase of ZnSO4 concentration by AAS determination. The effect of indution is remarkable. The production of MT-like reaches 2. 1ing per gram wet weight when the concentration of ZnSO4 is 1 mmol?L~-1.

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.

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.

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.

The N-terminal of porcine parvovirus (PPV) viral protein 2 (VP2) links a glycine-rich domain which is a cleavage site of PPV VP3.In order to confirm that the glycine-rich domain was essential for the self-assembling of virus-like particles (VLPs).The VP2 gene with glycine-rich domain deleted and the complete VP2 gene were inserted to eukaryotic expression vector pCI-neo and were named pCI-AVP2 and pCI-VP2. Then, pCI-delta VP2, pCI-VP2 and pCI-neo were transferred into Vero Cells by liposome and the VLPs was detected by SDS-PAGE, Western blotting, indirect immunofluorescence and immunoelectron microscopy. Furthermore, 56 female Kunming mice were divided into 5 groups and injected intramuscularly with pCI-delta VP2, pCI-VP2 and pCI-neo as DNA vaccine, PPV inactivated vaccine and normal saline separately. The peripheral blood of the mice was collected to analyze the subgroups of the peripheral blood mononuclear cell by flow cytometry, to detect the antibody and lymphocyte proliferation by indirect-ELISA and MTT assay separately. The results show that the VLPs were observed both in the pCI-delta VP2 and pCI-VP2 transferred Vero Cells. The two VLPs could agglutinate guinea pig erythrocytes. The results also show that both the pCI-delta VP2 and pCI-VP2 vaccine induced special cellular and humoral immunity effectively. Those results revealed that the glycine-rich domain is not essential for the VPL's self-assembling. This study provides a new theoretical evidence for the relationship between the gene structure and protein function of VP2.
Animals ; Antigens, Viral ; genetics ; metabolism ; Capsid Proteins ; genetics ; metabolism ; Cercopithecus aethiops ; Female ; Genetic Vectors ; genetics ; Glycine ; Mice ; Sequence Deletion ; Swine ; Transfection ; Vaccination ; Vaccines, Virus-Like Particle ; biosynthesis ; immunology ; Vero Cells

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.