[Clinicopathological and molecular genetic characteristics of ELOC mutated renal cell carcinoma].

Objective: To investigate the clinicopathological features, molecular genetic features, differential diagnosis and prognosis of ELOC mutated renal cell carcinoma. Methods: From January 2015 to June 2022, 11 cases of renal cell carcinoma with clear-cell morphology, expression of CAⅨ and CK7 and no 3p deletion were collected. Two cases of ELOC mutant renal cell carcinoma were diagnosed using whole exome sequencing (WES). The clinical features, morphology, immunophenotype, FISH and WES results were analyzed. The relevant literature was reviewed. Results: The two patients were both male, aged 29 and 51 years, respectively. They were both found to have a renal mass by physical examination. The maximum diameters of the tumors were 3.5 cm and 2.0 cm, respectively. At the low magnification, the tumors were well-defined. The tumor cells showed a pushing border and were separated by thick fibrous bands, forming nodules. The tumor cells were arranged in a variety of patterns, including tubular, papillary, solid nest or alveolar. At high magnification, the tumor cells were large, with well-defined cell borders and clear cytoplasm or fine eosinophilic granules. CAⅨ was diffusely box-like positive in both cases. Case 1 was partially and moderately positive for CK7, strongly positive for CD10, diffusely and moderately positive for P504S, and weakly positive for 34ßE12. In case 2, CK7 and CD10 were both partially, moderately positive and P504s were diffusely positive, but 34ßE12 was negative. FISH results showed that both cases had no 3p deletion. ELOC c.235T>A (p.Y79N) mutation was identified using WES in case 1, while ELOC c.236_237inv (p.Y79C) mutation was identified in case 2. Conclusions: As a new clinical entity, ELOC mutated renal cell carcinoma may be underdiagnosed due to its overlap with clear cell renal cell carcinoma in morphology and immunophenotype. The diagnosis of renal cell carcinoma with ELOC mutation should be confirmed by morphology, immunohistochemistry, FISH and gene mutation detection. However, more additional cases are needed to explain its biological behavior and prognosis.

[Role of microbial-derived antioxidants on diquat-induced oxidative stress, endoplasmic reticulum stress and function in mice].

Objective: To study the role of microbial-derived antioxidants (MA) based on the model of diquat-induced oxidative stress, endoplasmic reticulum stress, apoptosis and function in mice. Methods: 18 female C57BL/6 mice with body mass of 16~18 g were selected and randomly divided into 3 groups with 6 mice in each group. After 22 days of feeding, model and antioxidant group mice were intraperitoneally injected with diquat solution and control group were injected with same amount of isotonic saline. The content of free radical, MDA, antioxidant enzyme activity, aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activity were detected according to the instructions of the kit. QRT-PCR was used to detect the expression of endoplasmic reticulum stress and apoptosis-related genes. One-way analysis of variance was used for data comparison between groups. Results: Hydrogen peroxide (H(2)O(2)) in the control group, model group and antioxidant group was (8.74 ± 1.38), (11.44 ± 1.01), (9.81 ± 0.98) mmol/g prot, respectively, and the difference between the groups was statistically significant (F = 7.640, P < 0.05). MDA content in the control, model and antioxidant group were (0.65 ± 0.07), (0.86 ± 0.18), (0.70 ± 0.05) nmol/mg prot, respectively, and the difference between the groups was statistically significant (F = 5.406, P <0.05). Aspartate aminotransferase (AST) activity in the model group (146.68 ± 4.29) U/gprot was significantly higher than control group (125.64 ± 15.69) U/gprot and antioxidant group (126.57 ± 1.82) U/gprot, F = 6.192, P < 0.05. Real-time quantitative PCR result showed that the relative expression of protein kinase R-like endoplasmic reticulum kinase (PERK) and activated transcription factor 6 (ATF6) genes in the model group was significantly higher than control group, which were 1.880 ± 0.442 and 1.800 ± 0.380, F = 7.702 and 10.815, and P < 0.05, respectively. Apoptosis-related gene expression results showed that the relative expression levels of caspase3 and caspase8 genes in the antioxidant group (1.136 ± 0.381 and 1.593 ± 0.407) was significantly lower than model group (1.572 ± 0.127 and 2.843 ± 0.973), (F = 12.800, 7.657, P < 0.05). Conclusion: Microbial-derived antioxidants can reduce diquat-induced liver oxidative stress, endoplasmic reticulum stress and hepatocyte apoptosis in mice, and thus improves liver function.

The Effect of Maternal Diet with Fish Oil on Oxidative Stress and Inflammatory Response in Sow and New-Born Piglets.

Pregnancy is an oxidative stress and immune challenge for the mother. Fish oil is rich in EPA and DHA, which can partly inhibit aspects of inflammation and restore antioxidant capacity. In the present study, we investigated the effect of maternal diet with fish oil during the late gestation period on oxidative stress and inflammatory response in sows and their progenies. Twelve second-parity sows were allocated equally into two groups. Sows were fed either the soybean oil diet (SD) or soybean oil+fish oil diet (FD) during the gestation period. The plasma of sows, cord blood, and new-born piglets were collected. Full-term placentas and livers of new-born piglets were also sampled. The activities of glutathione peroxidase (GSH-Px) and total superoxide dismutase (T-SOD) in the plasma of sows on farrowing day were higher, and the concentrations of interleukin-6 (IL-6) and prostaglandin-endoperoxide synthase 2 (PGE2) in the plasma of sows on farrowing day and interleukin-1ß (IL-1ß) in the plasma of cord blood were lower in the FD group than those in the SD group (P < 0.05). The FD downregulated the expression of SOD, IL-1ß, IL-6, and transforming growth factor-ß-activated kinase 1 binding protein 1 (TAB1) mRNA but upregulated the expression of lipoxygenase enzyme 5 (ALOX5) and interleukin-10 (IL-10) mRNA in placentas (P < 0.05). The FD downregulated the protein expression level of p-JNK/JNK in placentas (P < 0.05). In the livers of new-born piglets, the FD upregulated the expression of ALOX5 (P < 0.05) and G-protein-coupled receptor 120 (GPR120) (P < 0.05) mRNA. Our results suggest that the maternal diet with fish oil might alleviate oxidative stress in sows on farrowing day and modulate inflammatory response in full-term placentas by inhibiting the JNK signal pathway. Moreover, the maternal diet with fish oil might partly regulate the neonatal immune response of their progenies.

[microRNA-146a reverses the inhibitory effects of Porphyromonas gingivalis lipopolysaccharide on osteogenesis of human periodontal ligament cells].

Objective: To investigate the effects and mechanisms of microRNA-146a (miR-146a) on osteogenic differentiation of human periodontal ligament cells (hPDLC) stimulated by lipopolysaccharide (LPS) of Porphyromonas gingivalis (Pg). Methods: hPDLC were cultured in vitro and induced to the phase of osteogenic differentiation. These cells were divided into five groups: non-osteogenic differentiation cells, osteogenic differentiation cells, osteogenic differentiation cells treated with Pg LPS, osteogenic differentiation cells treated with Pg LPS and miR-146a mimic, osteogenic differentiation cells treated with Pg LPS and miR-146a negative control. Osteogenic markers and mineralization were detected via quantitative real-time PCR (qPCR) and alizarin red staining, respectively. Meanwhile, non-radioactive transcription factor assay was applied to explore the nuclear activity of nuclear factor kappa B (NF-κB) p65 in nuclear extracts of hPDLC. Results: Compared with cells of osteogenic differentiation in non-LPS-stimulated groups, Pg LPS could decrease the markers of osteogenic differentiation of hPDLC such as collagen Ⅰ (Col-Ⅰ), alkaline phosphatase (ALP), Runt-related transcription factor-2 (RUNX2) and osteocalcin (OCN) (P<0.05), inhibit mineralization, and stimulate NF-κB p65 nuclear activity expression (non-LPS stimulated group: 1.023±0.217, LPS stimulated group: 6.252±0.613, P=0.008). However, compared with cells in Pg LPS/miR-146a negative control group, miR-146a increased Col-Ⅰ (P=0.007) and OCN (P=0.049) mRNA expression, rather than ALP (P=0.167) and RUNX2 (P=0.580) at day 3; miR-146a also upregulated mRNA levels of Col-Ⅰ, ALP, RUNX2 and OCN (P<0.05) at day 7 and day 14, and enhance mineralization. Meanwhile, miR-146a mimic could decrease the nuclear activity of NF-κB p65 induced by Pg LPS in hPDLC (miR-146a: 2.427±0.354, negative control: 5.863±0.482, P=0.019). Conclusions: miR-146a could reverse the inhibitory effects of Pg LPS on osteogenic differentiation of hPDLC through enhancing the expression of osteogenic markers and decreasing inflammatory pathway in hPDLC.

[Clinicopathologic features of adenoid cystic carcinoma of breast].

Objective: To investigate the clinicopathologic features and grading of adenoid cystic carcinoma (ACC) of the breast. Methods: Sixteen cases of ACC of the breast were analyzed and graded according to the previous report. Immunohistochemical (IHC) staining was used to detect the immunophenotype, Ki-67 proliferative index and expression of EZH2, and the association with tumor grade and outcome was analyzed. Results: Of the 16 cases, 11 were grade Ⅰ, with the epithelial and myoepithelial cells being arranged into tubular and cribriform structure with no solid component; three were grade Ⅱ, which were composed of mixed tubular, cribriform and solid component (<30%); and two were grade Ⅲ, which showed mainly solid component (>90%) and the tumor cells showed basaloid features with scanty cytoplasm and hyperchromatic nuclei, and mitotic count was>5/10 HPF. Immunophenotypically, the epithelial cells expressed CK7, CK8/18 and CD117; the myoepithelial cells expressed p63 and CK5/6; while the basaloid cells were positive for CK5/6 and CD117.Tubular and cibriform ACC showed low Ki-67 and EZH2 expression, while the two cases of solid variant with basaloid features showed high level of Ki-67 and EZH2 expression. Follow-up data were available in 13 cases with a median follow-up period of 42 months. Lung metastasis occurred after 12 months in one grade Ⅱ case and the patient died of disease after 34 months. Vertebral metastasis occurred after 12 months in one grade Ⅲ case and axillary lymph node metastasis occurred in another grade Ⅲ case. All other patients were free of disease at the end of the follow-up periods. Conclusions: ACC shows morphologic spectrum varying from low to high grade, the latter can may give rise to local and distant metastasis. ACC should not be regarded simply as low malignant potential, and should be graded for optimal treatment.

[Developmental carcinogenesis in the cheek pouch of the hamster induced by DMBA using on light microscope]

OBJECTIVE:In this study that the DMBA was applicated to induce the premalignantal changes of the cheek pouches of 89 hamsters in various stage,the result was shown that the slight epithelial displasia was occured 3 weeks later,the medium displasia was occured 5 weeks later,and the severe displasia or the carcinoma in situ was occured 6-9 weeks later.It is suggested that the suitable experimental period of the hamster's carcinogenesis is during 1-3 weeks.

[The study of influencing cellular dynamics of gynostemma pentaphyllum on golden hamster cheek pouch premalignancy]

Studying premalignancy is an important field of conquering cancer.The aim to study the mechanics of oral mucosal premalignant formation and seek effective drug which could prevent and block cancer,Gynostemma Pentaphyllum(GP) was applies to block carcinomatous conversion of golden hamster cheek pouch by Salley's way.Using light microscopic observation and proliferating cell nuclear antigen(PCNA) as well as bromodeoxyuridine(BrdU)(PCNA AND BrdU are immunohistochemistry techniques),GPs effect was observed dynamically.We found that GP could delay the presence of leukoplakia and decrease the rate of presence as well as cancer-causing rate. The positive cells of taking drug group PCNA and BrdU were lower than model group significantly and in a lower condition all the time.It is illustrated that GP has the effect of preventing cancer and inhibiting cellular dysplasia.

[The study of cellular dynamic varied features on golden hamster cheek pouch premalignant model]

Researching the model of golden hamster cheek pouch premalignancy not only had significance on treating and preventing oral epithelial cancers,but also could furnish the reference material about other similar hidden tumors.In this study,immunohistochemistry ways(proliferation cell nuclear antigen-PCNA,bromodeoxyurdine-BRDU) combined with light and electric microscopic observation were applied to study cellular dynamic varied features on golden hamster cheek pouch premalignant model induced by dimethylbenzanthracence (DMBA).The results showed that both S-period cells and DAN's synthesis increased in the development of premalignancy,then,discovered the basis of cellular dynamics of the model.

[Correlative analysis with proliferating cell nuclear antigen,bromodeoxyuridine,and microscopic observation on golden hamster carcinogenesis model]

The early diagnosis of carcinomatous conversion has special significance on increasing effectiveness.Proliferating cell nuclear antigen(PCNA),bromodeoxyuridine(BRDU)(immunohistochemistry ways),and microscopic diagnosis were applied and observed on golden hamster cheek pouch carcinogenisis model induced by dimethybenzanthracene(DMBA) in order to seek simple,effective,leading and reliable diagnostic way.The results showed that three observable means had significant correlation,and demonstrated that immunohistochemistry ways practical value in diagnosis of carcinomatous conversion.This study suggests that PCNA and BRDU are more sensitive,simple and reliable,and two hopeful methods of early diagnosis of carcinomatous conversion.

[Correlative analysis with proliferating cell nuclear antigen and bromodeoxyuridine on golden hamster carcinogenesis model]

The early diagnosis of carcinomatous conversion has special significance on increasing therapeutic effectivencess.Proliferating cell nuclearnatigen(PCNA),bromodexoyuridine(UrdU)(immunohistochemistry ways),and microscopic diagnosis were applied and observed on golden hamster cheek is pouch carcinogenisis model induced by dimethylbenzanthracene(DMBA) in order to seek simple,effective,leading and reliable diagnostic way.The results showed that three observable means had significant correlativity,and demonstrated that immunohistochemistry ways have practical value in diagnosis of carcinomatous conversion.This study suggests that PCNA and BrdU be more sensitive,simple and reliable,and hopeful methods of early diagnosis of carcinomatous conversion.

[Teatment of oral lichen planus with chloroquine and triamcinolone-A]

In this study,single blind grouping method was applied to compare the effect of chloroquine and triamcinolone-A which were used to treat 360 cases of oral lichen planus.The results showed the effect of local injecting triamcinolone-A was better than that of taking chloroquine,and also was better than that of taking triamcinolone-A accompained of chloroquine.Therefore,they were two effective ways to cure OLP and worthy of wide use.

[The clinical analysis of 39 cases of oral mucosal hyperplasia]

This study was reviewing in analysed the clinical,histopathological and treatment features of 39 cases suffered in oral mucosal hyperplasia.It definited the sorts of disease which may be malignant changed and the histopathological grading criteria of hyperplasis.The different treatment methods were administered according to the different grade of the malignant change.The results of the post-treatment observation showed that the drug method only be fit for the cases suffered slight hyperplasia without nuclei differentiation.

[The application of Immuno-gold and silver stainning to detecting OLP and DLE]

Immuno-gold and silver staining (IGSS) is an advanced labelling method followed by Immuno-fluorescence(IF),Immunoenzyme and Radio-immunity.In this study,the IGSS method for detection of deposits of Immunogloblins (IgG,IgA,IgM) at the BMZ had been performed on biopsy specimens from oral lesion of patients with OLP and DLE.The results indicated that the sensitivity and specificity of IGSS were super than that of DIF.It suggested the IGSS be an available method to detect oral mucosal diseases widely.