Objective:To investigate the value of soluble interleukin-6 (IL-6) receptor (sIL-6R) level in predicting ultrafiltration insufficiency in peritoneal dialysis (PD) patients.Methods:It was a prospective cohort study. The patients who received continuous ambulatory PD and regular follow-up between November 2016 and July 2018 in the PD Center of Renji Hospital, School of Medicine, Shanghai Jiao Tong University were enrolled. Enzyme-linked immunosorbent assay was used to determine dialysate sIL-6R and its appearance rate (AR) was calculated. Patients were divided into high sIL-6R AR group and low sIL-6R AR group according to median value of sIL-6R AR and prospectively followed up until death, PD cessation, or the end of the study (December 31, 2022). Multiple linear regression was used to analyze the related factors of sIL-6R AR. Kaplan-Meier method and log-rank test were used to compare the survival rate difference of ultrafiltration insufficiency between high sIL-6R AR group and low sIL-6R AR group. Multivariate Cox regression and multivariate competing risk models were used to assess the risk factors associated with occurrence of ultrafiltration insufficiency.Results:A total of 198 PD patients were enrolled, including 115 (58.1%) males, with age of (54.9±13.7) years old and PD duration of 22.5 (6.6, 65.0) months. The sIL-6R AR of the cohort was 2 094.7 (1 672.4, 2 920.9) pg/min. Compared with low sIL-6R AR（<2 094.7 pg/min）group, high sIL-6R AR（>2 094.7 pg/min）group had older age ( t=-3.269, P=0.001), higher body mass index ( t=-3.248, P=0.001), proportion of combined diabetes mellitus ( χ2=8.890, P=0.003), 24 h glucose exposure ( Z=-2.257, P=0.024), 24 h ultrafiltration capacity ( Z=-2.515, P=0.012), 4 h dialysate creatinine to serum creatinine ratio ( t=-2.609, P=0.010), mass transfer area coefficient of creatinine ( Z=-2.308, P=0.021), IL-6 AR ( Z=-3.533, P<0.001) and solute glycoprotein 130 AR ( Z=-8.670, P<0.001), and lower serum albumin ( t=2.595, P=0.010) and residual renal function ( t=2.133, P=0.033). Multiple linear regression analysis showed that body mass index ( β=0.194, P=0.005), serum albumin ( β=-0.215, P=0.002) and dialysate lg[IL-6 AR] ( β=0.197, P=0.011) were independently correlated with sIL-6R AR. By the end of the study, 57 (28.8%) patients developed ultrafiltration insufficiency. Kaplan-Meier analysis showed that high sIL-6R AR group had a significantly inferior ultrafiltration insufficiency-free survival rate than that in low sIL-6R AR group (log-rank χ 2=5.375, P=0.020). Multivariate Cox regression analysis and multivariate competing risk models showed that high dialysate sIL-6R AR (>2 094.7 pg/min) was an independent influencing factor of ultrafiltration insufficiency ( HR=2.286 , 95% CI 1.254-4.165 , P=0.007 ; SHR=2.074, 95% CI 1.124-3.828, P=0.020) in PD patients. Conclusions:Dialysate sIL-6R level was associated with body mass index, serum albumin and dialysate IL-6 level. Dialysate sIL-6R may be a predictive factor of ultrafiltration insufficiency in PD patients.

Purpose To analyze the precursor lesions,clinicopathological features and prognosis of mucinous carcinoma(MC).Methods A total of 303 MC cases diagnosed by surgi-cal specimens were included,including 193 pure mucinous car-cinomas(PMC)and 110 mixed mucinous carcinomas(MMC).PMC included 163 cases of type A(hypocellular type)and 30 cases of type B(hypercellular type).The histomorphological characteristics,surrounding breast tissue morphology,and im-mune markers were evaluated to analyze the clinicopathological features related to MC precursor lesions and prognosis.Results The median age at diagnosis of MC was 50 years,and the me-dian tumor size was 2.5 cm.Compared to PMC,MMC had more lymph node(LN)involvement,more grade Ⅲ-Ⅳ and Ki67＞20％tumors(P＜0.05).There were no significant differences in tumor size,LN involvement and clinical stage between type A and type B PMC.55 cases of MC(49 PMC and 6 MMC)with precursor lesions of mucocoele-like lesions(MLL)were all grade 1 to 2,most of them(54/55,98.2％)were T1-T2,and the proportions of MC component in all 6 MMCs were all ≥ 50％.There were 119 cases of MC whose precursor lesions were ductal carcinoma in situ(DCIS),of which 28.6％of cases(34/119)had LN involvement,and 16.8％(20/119)of cases showed high proliferative activity(Ki67＞20％).The precursor lesions of MC in 16 cases were solid papillary carcinoma(SPC)in situ,inclu-ding 7 cases of type BPMC and 9 cases of MMC.The median age of these patients was 67 years and no recurrence or metastasis was observed during follow-up.The disease-free survival(DFS)and overall survival(OS)of the 303 cases of MC was 93.5％and 98.6％,respectively.PMC had a better prognosis than MMC(DFS:95.2％vs.90.6％;OS:99.5％vs.97.2％),but there was no significant difference between type A and type B PMC.The prognosis of patients with LN involvement and clinical stage Ⅲ-Ⅳ was worse(P＜0.05).Conclusion MC of the breast is a kind of heterogeneous malignant tumors.Our findings support that MC with specific precursor lesions have different ev-olutionary pathways,that MC with precursor lesions of MLL and SPC have a good prognosis,and MC associated with high-grade DCIS are more aggressive.LN involvement,higher clinical grade,and younger age are associated with poor prognosis.Focu-sing on precursor lesions and high risk clinicopathological fea-tures can contribute to providing more effective treatments for these patients.

Purpose To analyze the precursor lesions,clinicopathological features and prognosis of mucinous carcinoma(MC).Methods A total of 303 MC cases diagnosed by surgi-cal specimens were included,including 193 pure mucinous car-cinomas(PMC)and 110 mixed mucinous carcinomas(MMC).PMC included 163 cases of type A(hypocellular type)and 30 cases of type B(hypercellular type).The histomorphological characteristics,surrounding breast tissue morphology,and im-mune markers were evaluated to analyze the clinicopathological features related to MC precursor lesions and prognosis.Results The median age at diagnosis of MC was 50 years,and the me-dian tumor size was 2.5 cm.Compared to PMC,MMC had more lymph node(LN)involvement,more grade Ⅲ-Ⅳ and Ki67＞20％tumors(P＜0.05).There were no significant differences in tumor size,LN involvement and clinical stage between type A and type B PMC.55 cases of MC(49 PMC and 6 MMC)with precursor lesions of mucocoele-like lesions(MLL)were all grade 1 to 2,most of them(54/55,98.2％)were T1-T2,and the proportions of MC component in all 6 MMCs were all ≥ 50％.There were 119 cases of MC whose precursor lesions were ductal carcinoma in situ(DCIS),of which 28.6％of cases(34/119)had LN involvement,and 16.8％(20/119)of cases showed high proliferative activity(Ki67＞20％).The precursor lesions of MC in 16 cases were solid papillary carcinoma(SPC)in situ,inclu-ding 7 cases of type BPMC and 9 cases of MMC.The median age of these patients was 67 years and no recurrence or metastasis was observed during follow-up.The disease-free survival(DFS)and overall survival(OS)of the 303 cases of MC was 93.5％and 98.6％,respectively.PMC had a better prognosis than MMC(DFS:95.2％vs.90.6％;OS:99.5％vs.97.2％),but there was no significant difference between type A and type B PMC.The prognosis of patients with LN involvement and clinical stage Ⅲ-Ⅳ was worse(P＜0.05).Conclusion MC of the breast is a kind of heterogeneous malignant tumors.Our findings support that MC with specific precursor lesions have different ev-olutionary pathways,that MC with precursor lesions of MLL and SPC have a good prognosis,and MC associated with high-grade DCIS are more aggressive.LN involvement,higher clinical grade,and younger age are associated with poor prognosis.Focu-sing on precursor lesions and high risk clinicopathological fea-tures can contribute to providing more effective treatments for these patients.

Tyrosol is a natural polyphenolic product that is widely used in chemical, pharmaceutical and food industries. Currently, the de novo synthesis of tyrosol by Escherichia coli suffers from issues such as low cell density and poor yield. Therefore, the phenylpyruvate decarboxylase mutant ARO10^F138L/D218G obtained in our previous study was fused with an alcohol dehydrogenase from different microorganisms for fusion expression, and the optimal ARO^{10F138L/D218G}-L-YahK produced 1.09 g/L tyrosol in shake flasks. In order to further improve tyrosol production, feaB, a key gene in the competing pathway of 4-hydroxyphenylacetic acid, was knocked out, and the resulted strain produced 1.26 g/L tyrosol with an increase of 21.15% compared to that of the control. To overcome the low cell density in tyrosol fermentation, the quorum-sensing circuit was used to dynamically regulate the tyrosol synthesis pathway, so as to alleviate the toxic effect of tyrosol on chassis cells and relieve the growth inhibition. Using this strategy, the yield of tyrosol was increased to 1.74 g/L, a 33.82% increase. In a 2 L fermenter, the production of tyrosol in the engineered strain TRFQ5 dynamically regulated by quorum-sensing reached 4.22 g/L with an OD₆₀₀ of 42.88. Compared with those in the engineered strain TRF5 statically regulated by induced expression, the yield was increased by 38.58% and the OD₆₀₀ was enhanced by 43.62%. The combination of blocking the competing pathway using gene knockout technology, and reducing the inhibitory effect of tyrosol toxicity on chassis cells through quorum-sensing dynamic regulation increased the production of tyrosol. This study may facilitate the biosynthesis of other chemicals with high toxicity.
Escherichia coli/genetics* ; Biological Products ; Bioreactors ; Fermentation

Hereditary gingival fibromatosis (HGF) is a rare inherited condition with fibromatoid hyperplasia of the gingival tissue that exhibits great genetic heterogeneity. Five distinct loci related to non-syndromic HGF have been identified; however, only two disease-causing genes, SOS1 and REST, inducing HGF have been identified at two loci, GINGF1 and GINGF5, respectively. Here, based on a family pedigree with 26 members, including nine patients with HGF, we identified double heterozygous pathogenic mutations in the ZNF513 (c.C748T, p.R250W) and KIF3C (c.G1229A, p.R410H) genes within the GINGF3 locus related to HGF. Functional studies demonstrated that the ZNF513 p.R250W and KIF3C p.R410H variants significantly increased the expression of ZNF513 and KIF3C in vitro and in vivo. ZNF513, a transcription factor, binds to KIF3C exon 1 and participates in the positive regulation of KIF3C expression in gingival fibroblasts. Furthermore, a knock-in mouse model confirmed that heterozygous or homozygous mutations within Zfp513 (p.R250W) or Kif3c (p.R412H) alone do not led to clear phenotypes with gingival fibromatosis, whereas the double mutations led to gingival hyperplasia phenotypes. In addition, we found that ZNF513 binds to the SOS1 promoter and plays an important positive role in regulating the expression of SOS1. Moreover, the KIF3C p.R410H mutation could activate the PI3K and KCNQ1 potassium channels. ZNF513 combined with KIF3C regulates gingival fibroblast proliferation, migration, and fibrosis response via the PI3K/AKT/mTOR and Ras/Raf/MEK/ERK pathways. In summary, these results demonstrate ZNF513 + KIF3C as an important genetic combination in HGF manifestation and suggest that ZNF513 mutation may be a major risk factor for HGF.
Animals ; Humans ; Mice ; Fibromatosis, Gingival/pathology* ; Gingiva ; Kinesins/genetics* ; Mutation/genetics* ; Phosphatidylinositol 3-Kinases/genetics*

Grape (Vitis vinifera L.) in production is frequently exposed to inadequate light, which significantly affects its agronomic traits via inhibiting their physiological, metabolic and developmental processes. To explore the mechanism how the grape plants respond to the weak light stress, we used 'Yinhong' grape and examined their physiology-biochemistry characteristics and transcriptional profile under different levels of weak light stress. The results showed that grape seedlings upon low intensity shading treatments were not significantly affected. As the shading stress intensity was strengthened, the epidermis cells, palisade tissue, and spongy tissue in the leaves were thinner, the intercellular space between the palisade tissue and spongy tissue was larger compared with that of the control, and the activities of superoxide dismutase, catalase and peroxidase were decreased gradually. Additionally, the soluble protein content increased and the free proline content decreased gradually. Compared with the control, significant changes in plant photosynthetic characteristics and physiology-biochemistry characteristics were observed under high intensity of shading (80%). RNA-seq data showed that the differentially expressed genes between CK and T2, CK and T4, T2 and T4 were 13 913, 13 293 and 14 943, respectively. Most of the enrichment pathways were closely related with the plant's response to stress. Several signaling pathways in response to stress-resistance, e.g. JA/MYC2 pathway and MAPK signal pathway, were activated under weak light stress. The expression level of a variety of genes related to antioxidation (such as polyphenol oxidase and thioredoxin), photosynthesis (such as phytochrome) was altered under weak light stress, indicating that 'Yinhong' grape may activate the antioxidation related pathways to cope with reactive oxygen species (ROS). In addition, it may activate the expression of photosynthetic pigment and light reaction structural protein to maintain the photosynthesis activity. This research may help better understand the relevant physiological response mechanism and facilitate cultivation of grape seedlings under weak light.
Vitis/metabolism* ; Gene Expression Regulation, Plant ; Photosynthesis/genetics* ; Plant Leaves ; Light ; Seedlings/metabolism*

Objective To investigate the effect of pyrin domain 3 (NLRP3) inflammasome in the process of contrast induced human kidney cell apoptosis.Methods Human kidney 2 (HK-2) cells were cultured in DMEM-F12 medium with 5％ FBS.Cells were divided into control group,Contrast group (O group),NLRP3-siRNA+Iohexol group (si-NLRP3+O group),ASC-siRNA+Iohexol group (si-ASC+O group),and mannitol group (M group).Different concentrations of hypotonic contrast agent were added to HK-2 cell culture plates for 24,48 and 72 h.Flow cytometry was used to detect apoptosis.NLRP3 and ASC mRNA expressions were detected by RT-PCR.The expressions of NLRP3,ASC,caspase-8/cleaved caspase-8,Bcl-2/Bax,caspase-1/cleaved caspase-1,and caspase-3/cleaved caspase-3 protein were detected by Western blot.The levels of interleukin (IL) 1β and IL-18 in supernatant were detected by ELISA.Results Compared with the control group,the rate of apoptotic cells,as well as the expressions of NLRP3,ASC and cleaved caspase-1 proteins were increased in HK-2 cells of contrast group.The expressions of NLRP3 and ASC mRNA in the contrast group also increased,so did IL-1β and IL-18 levels (all P＜0.05),suggesting that NLRP3 inflammasome in HK-2 cells was activated by contrast.Compared with the control group,the expressions of cleaved caspase-8,Bax and cleaved caspase-3 protein were increased,and the expression of anti-apoptotic protein Bcl-2 was decreased (all P ＜ 0.05).Compared with the contrast group,the rate of apoptotic cells in the si-NLRP3 + contrast group and si-ASC + contrast group was significantly decreased;the expression of cleaved caspase-1 was decreased;the expressions of Bax and cleaved caspase-3 were decreased,and Bcl-2 level was increased.The expressions of IL-1β and IL-18 in the supernatant of cells were decreased (all P ＜ 0.05).Conclusion Contrast agent can activate the NLRP3 pathway in HK-2 cells and induce apoptosis,which could be reduced by blocking the NLRP3 pathway.

Recently, a novel atypical porcine pestivirus (APPV) in pig was reported. In this study, two APPV strains, APPV-China/GZ01/2016 (GZ01) and APPV-China/GD-SD/2016 (GD-SD), were identified in two newborn piglet herds with congenital tremor from China. The open reading frame of the two strains shared an 83.5% nucleotide identity. Phylogenetically, the APPV strains were placed into two groups: GZ01 belonged to group I and GD-SD belonged to group II. A high viral load was detected in the cerebellum (quantification cycles < 26). Further studies should be carried out to thoroughly elucidate the development of congenital tremors caused by APPV.
Cerebellum ; China ; Genome ; Humans ; Infant, Newborn ; Open Reading Frames ; Pestivirus ; Tremor ; Viral Load

Objective To clarify the role of pentraxin 3 (PTX3) in the development of peripheral arterial disease (PAD) in maintenance hemodialysis (MHD) patients. Methods One hundred and sixteen patients undergone MHD therapy in our center for more than 3 months were enrolled in the study. Clinical data were collected for analysis. Ankle-brachial index (ABI) was used to estimate the presence of PAD. Patients were divided into PAD group (ABI<0.9) and nonestimate the association of PAD with PTX3 as well as other potential risk factors. Results The incidence of PAD was 18.1% (21/116). Plasma level of PTX3 was significantly higher in PAD patients than that in non-PAD patients [(5.55 ±2.63) μg/L vs (2.32 ±1.29)μg/L, P<0.01].Univariate analysis showed ABI values were negatively correlated with plasma PTX3 levels (r =-0.548, P<0.01), high-sensitivity C-reactive protein (hsCRP), age, blood glucose and triglyceride. ROC curve of PAD revealed that area under curve (AUC) of PTX3 was 0.901 (P<0.01). With the cut-off value of PTX3 as 4.06 μg/L, the diagnostic sensitivity and specificity in PAD were 81.0% and 91.5%. ROC curve of PAD showed that AUC of hsCRP was 0.640 (P<0.05). With the cut-off value of hsCRP as 3.33 mg/L, the diagnostic sensitivity and specificity in PAD were 57.1% and 56.8%. Using Logistic regression, plasma PTX3 was found to be associated with PAD (0R=9.755, 95%CI:2.359-19.354, P=0.001). Conclusions The PAD incidence of MHD patients in our center is 18.1%. Plasma PTX3 level is significantly correlated with the presence of PAD in MHD patients. The sensitivity and specificity of PTX3 are higher than those of hsCRP for PAD diagnosis.