Objective To explore the therapeutic effects and the mechanisms of Faecalibacterium prausnitzii (Fp) on trinitro-benzene-sulfonic acid-induced colitis.Methods Sixty Sprague-Dawley rats were divided into healthy control group, colitis model control group, Fp pretreated group,Fp supernatant pretreated group,Fp treated group and Fp supernatant treated group.Disease activity index (DAI),histological injury of colonic tissue,the content of butyrate in feces,forkhead box protein 3 (Foxp3) regulatory T cells (Treg) in peripheral blood and spleen and the level of interlenkin (IL)-17 and IL-6 in serum were evaluated.All the data were statistical analyzed by single factor analysis of variance. Results Compared with colitis model control group, DAI significantly lowered and histological injury obviously improved in Fp pretreated group, Fp supernatant pretreated group,Fp treated group and Fp supernatant treated group.The effects of Fp pretreated group were better than those of Fp treated group and Fp supernatant pretreated group were better than Fp supernatant treated group.The concentration of butyrate in Fp pretreated group,Fp supernatant pretreated group,Fp treated group and Fp supernatant treated group was (3091.08 ±485.50) × 106 mol/L,(1714.64 ± 351.25) × 10(-8) mol/L,(2064.75 ± 295.04) × 10-6 mol/L and (1089.13±321.23) × 10-6 mol/L respectively,there was significant difference between Fp pretreated group and other groups (F=49.796,P＜0.01).The peripheral blood level of Foxp3+ Treg in Fp supernatant pretreated group was highest.The spleen level of Foxp3+ Treg in Fp pretreated group and Fp supernatant pretreated group were significantly higher than that of other groups.The serum level of IL-17 and IL-6 in Fp pretreated group,Fp supernatant pretreated group,Fp treated group and Fp supernatant treated group was significantly lower than that of colitis model group.Conclustons Fp plays a role in promoting the repair of intestinal inflammatory reaction in colitis model rats.The mechanism may be related with butyrate producing,the peripheral blood and spleen level of Foxp3+ Treg up-regulating,suppressing the secretion of proinflammatory cytokine IL-17 and IL-6.Rebuilding the balance of Treg/Th17 to reduce local intestinal inflammation.

Objective To investigate the protective effects and mechanism of Faecalibacterium prausnitzii (Fp) and its products in 2,4,6-trinitrobenzene sulfonic acid (TNBS) induced colitis rats.Methods A total of 40 Sprague-Dawley rats were divided into healthy control group,colitis model group,Fp supernatant group,Fp bacteria group and Bifidobacterium longum (B.longum) group.The rats of the later four groups were enemaed with TNBS to establish the model.At five days before and one day after modeling,the rats were gavaged with phosphate buffer saline (PBS),the supernatant of Fp,live Fp bacteria and live B.longum respectively.Rats were executed at 48 hour after modeling.The colon tissues were taken for pathology examination.The content of short-chain fatty acids (SCFA) in fecal was tested by gas chromatography.The plasma level of interleukin-10 (IL- 10),interleukin-12 ( IL-12),interleukin-17 (IL-17) and interleukin-23 (IL-23) were detected by enzyme-linked immunosorbnent assay (ELISA) and the expression of IL-17 in intestinal mucosal tissues was measured by immunohistochemistry.The data were analyzed by one way ANOVA.Results Compared with the healthy control group,the rats of colitis group suffered serious weight loss and their intestinal pathology score increased [(193.57±14) g vs (170.25±19.18) g,(1.00±0.99) vs (3.34±0.38),t=2.83 and 7.55,all P value＜0.05].The Fp supernatant group showed protective effects in terms of weight and intestinal pathology score [(187.00± 14.67) g,(2.50±0.44),t=2.1 and 2.9,all P＜0.05].Compared with healthy control group,the plasma and colon tissue IL-17 concentration of colitis model group increased (16.61 pg/ml±2.45 pg/ml vs 20.47 pg/ml± 1.45 pg/ml,0.83±0.98 vs 5.14±0.90) (all P＜0.05).Compared with the colitis model group,the plasma and colon tissue IL-17 concentration of Fp supernatant group decreased ( 17.54 pg/ml± 1.51 pg/ml and 2.86±0.69).Conclusion Fp can regulate immune response and suppress rat colonic inflammation,which may be related with the expression of IL-17.

AIM: To investigate the mechanism of berberine’s inhibiting growth and metastasis of tumor by observing the effects of berberine on proliferation and apoptosis of human umbilical vein endothelial cells (HUVECs). METHODS: Cultured HUVECs were incubated with berberine. MTT assay and quantitative immunocytochemistry were used to detect cell proliferation and the expression of proliferation cell nuclear antigen (PCNA). Morphologic changes of apoptosis were observed by fluorescent staining, and Rhodamin123 was used to determinate mitochondrial membrane potential under the laser confocal microscopy. RESULTS: HUVEC proliferation was inhibited by co-incubating with berberine (20 mg/L) for 24 h and berberine (10 mg/L, 20 mg/L) for 48 h (P

Objective:To examine the correlations of microRNA-34c(miR-34c) expression in the peripheral blood with the onset of diabetic foot ulcer(DFU)and diabetic foot osteomyelitis(DFO)in patients with type 2 diabetes mellitus(T2DM).Methods:Sixty newly-diagnosed T2DM patients without DFU(T2DM group), 112 T2DM patients with DFU(DFU group), and 60 healthy controls with normal glucose tolerance(NC group)were included. The 112 T2DM patients with DFU were further divided into DFO( n=64)and NDFO( n=48)groups. The levels of miR-34c were determined by quantitative real-time PCR, while clinical features and risk factors of DFU and DFO were explored. Results:A significant increase in the expression level of miR-34c in peripheral blood was observed in T2DM group compared with NC group[2.99(1.45-6.22) vs 1.01(0.89-1.52), P<0.05], and a markedly increased miR-34c expression level was noted in DFU group compared with T2DM group [9.65(6.15-18.63) vs 2.99(1.45-6.22), P<0.01]. Additionally, the expression level of miR-34c in peripheral blood significantly increased in DFO group compared to NDFO group [13.46(8.89-19.11) vs 6.02(5.93-14.72), P<0.01]. Moreover, there was a positive correlation between the expression level of miR-34c in peripheral blood and the amputation rate in patients in DFU group( P=0.030), and a negative correlation in the expression level of miR-34c( P=0.025)with healing rate of DFU after eight weeks. The multivariate logistic regression analysis confirmed that a high expression of miR-34c was an independent risk factor for DFU and DFO( OR=3.52, 4.13; both P<0.01). Conclusion:An increased expression of miR-34c in peripheral blood of T2DM patients might be closely related to the occurrence, development, and prognosis of DFU and DFO.

A 44-year-old pregnant woman (G5P3) who had delivered two children with DMD was admitted and underwent prenatal diagnosis at Peking Union Medical College Hospital in 2019. (1) The karyotype of the fetus in 2019 was 47,XXY. The fluorescence in situ hybridization (FISH) result showed a nucish(CSPX×2, CSPY×1)[100] and multiplex ligation-dependent probe amplification (MLPA) suggested sex chromosome abnormality. Based on the above results, the fetus was diagnosed with Klinefelter syndrome. Fetal short tandem repeat (STR) linkage analysis and Sanger sequencing indicated a heterozygous mutation of c.9543delG(p.Trp3181CysfsTer2). (2) Sanger sequencing of the proband found a novel frameshift mutation of c.9543delG(p.Trp3181CysfsTer2 ) in exon 65 of the DMD gene. (3) The male fetus performing prenatal diagnosis in 2008 was found to have the same maternal gene markers as the proband with the same genotype. While the genotype of the fetus in 2009 obtained a different maternal gene marker from the proband and did not detect the same DMD gene mutation. This fetus was delivered at full term and was good during follow-up. (4) The elder brother and cousin of the proband had the same frameshift mutation in exon 65 of the DMD gene as the proband. The mother of the proband was a heterozygous carrier of the mutation.

Objective] To amplify and sequence the light chain of anti idiotypic monoclonal antibody NP30 of Schistosoma japonicum. [Methods] By comparing the conserved regions at each end of the nucleotide sequences of murine germ line genes enco ding FR1 and FR4 regions of immunoglobulin light chain variable regions, we designed a set of primers for amplification of V L gene. The hybridoma cells secreting anti idiotypic monoclonal antibody NP30 of Schistosoma japonicum were cultured and their genome DNAs were extracted and used as templates for PCR. The PCR product was then cloned into pUC19 vector. The recombinants were sequenced by Sanger′s method. The V L gene was compared with GenBank and published mouse V L genes. [Results] The full length of V L gene was 318 bp. The V L gene was a member of mouse Ig ? light chain subgroup IV and generated from rearrangement of germ line V and J? 4 genes. The V L gene sequence has been registered by GenBank(accession No. AF206720). [Conclusion] The obtained V L gene was a potentially functional gene of anti idiotypic monoclonal antibody NP30 of Schistosoma japonicum .

Objective To analyze the molecular epidemiology of adenovirus ( ADV) causing acute respiratory diseases and to investigate the mixed infection of ADV and other respiratory pathogens in Hebei Province. Methods Sputum samples were collected from inpatient children with acute respiratory diseases at Children′s Hospital of Hebei Province between June 2017 and May 2018. Multiplex reverse transcription PCR assay was used to detecte 13 kinds of respiratory pathogens. Nested PCR was performed to amplify ADV hexon gene and the amplified products were then sequenced. Results A total of 353 ADV-positive speci-mens were detected in 8839 specimens with a positive rate of 3. 99％. Significant difference in the positive rate of ADV was not observed between male and female patients (χ2=0. 0003, P=0. 99), but found among different age groups (χ2=115. 69, P<0. 001). All isolated ADV strains belonged to 11 serotypes, which were type 1 (16. 15％, 57/353), type 2 (35. 98％, 127/353), type 3 (21. 25％, 75/353), type 4 (1. 13％, 4/353), type 5 (11. 33％, 40/353), type 6 (3. 97％, 14/353), type 7 (8. 22％, 29/353), type 31 (0. 28％, 1/353), type 41 (0. 28％, 1/353), type 55 (0. 28％, 1/353) and type 57 (1. 13％, 4/353). Among the 353 ADV-positive specimens, 259 were mixed infections mainly caused by ADV and human rhinovirus (35. 52％). ADV and respiratory syncytial virus co-infections accounted for 12. 74％ and 33. 20％ of the mixed infections involved three or more pathogens. ADV could be detected throughout the year, especially in September and April to May. The predominant serotypes were types 1, 2 and 3. The av-erage ages of the two groups of ADV infection alone and ADV mixed infection were (27. 56±24. 67) months and (21. 33 ±20. 28) months, respectively, and the difference between them was statistically significant (P=0. 037). The incidence of ADV 2 infection alone was 25. 77％ (25/97), which was lower than that of ADV 2-involved mixed infection [39. 84％ (102/256),χ2=6. 05, P=0. 014]. However, the rate of ADV 7 infection alone was significantly higher than that of ADV 7-involved mixed infection [16. 49％ 16/97) vs 5. 08％ (13/256),χ2=6. 05, P<0. 001]. Conclusion ADV 1, ADV 2 and ADV 3 were the predominant serotypes circulating in Hebei Province from June 2017 to May 2018, especially in September and April to May. The younger the patients were, the higher the incidence would be. ADV 2 was prone to cause mixed infections with other respiratory pathogens, while ADV 7 was less common in mixed infections. Younger pa-tients were more susceptible to mixed infections. The most common co-infection was caused by ADV and hu-man rhinovirus.

Objective:To investigate the effects of terlipressin (TP) combined with norepinephrine (NE) on liver function and prognosis of patients with septic shock.Methods:From June 2018 to December 2019, 96 patients with septic shock and liver function impairment admitted to the ICU of Tianjin First Central Hospital were selected for prospective study. The patients were divided into control group( n=48) and experiment group( n=48) by randomize number table derived by computer. Based on conventional treatment, NE was used in control group, and the low dose continuous infusion of TP combined with NE was used in experiment group. Serial measurements of aspartate aminotransferase (AST), alanine aminotransferase (ALT), serum total bilirubin (TBIL), heart rate and mean arterial pressure (MAP), and blood lactic acid levels were made before the treatment and after the treatment at 24 and 48 hours. The mechanical ventilation time, intensive care unit (ICU) stay, and total length of hospital stay of the two groups were compared, and the 28-day mortality and serious adverse reactions of the two groups was also calculated. Results:The levels of ALT, AST, TBIL, heart rate and blood Lac of the two groups were significantly decreased after the treatment (all P<0.01), and the level of MAP was significantly increased (all P<0.01). Compared with the control group, the levels of 24-hour and 48-hour ALT, AST, TBIL, blood Lac of the experiment group were significantly decreased (all P<0.05), and the 48-hour level of MAP was significantly increased (all P<0.05), but there was no statistically significant difference between the two groups in the levels of 24-hour heart rate and 24-hour MAP (all P>0.05). Besides, there was no statistically significant difference between the two groups in the mechanical ventilation time, ICU stay, total length of hospital stay and the 28-day mortality (all P>0.05). And there were no serious adverse reactions such as avascular necrosis of the fingers and myocardial infraction in the two groups. Conclusions:In the treatment of septic shock, on the basic of adequate fluid resuscitation, continuous intravenous pumping of low-dose TP combined with NE can play a certain protective effect on the live, and the mechanisms of action may be mediated by stabilizing hemodynamics, reducing heart rate, reducing the level of blood Lac and improving liver perfusion, thereby protecting liver function in patients with septic shock.

Objective: To study the clinicopathological features, immunohistochemical phenotype, molecular changes, differential diagnosis and prognosis of eosinophilic solid and cystic renal cell carcinoma (ESC RCC). Methods: A total of 15 cases were selected from 2005 to 2019 at Nanjing Jinling Hospital,Nanjing University School of Medicine for clinicopathological and immunohistochemical analysis, 10 of which were subject to cancer-associated mutation analysis using targeted next-generation sequencing (NGS) panel. A literature review was also performed. Results: The patients′ ages ranged from 15 to 68 years (mean, 33 years). The male-to-female ratio was 1.1∶1.0. During a mean follow-up of 22 months, none of the patients developed tumor recurrence, progression or metastasis. Histologically, the tumors typically demonstrated solid and cystic architectures and the neoplastic cells contained voluminous eosinophilic cytoplasm with prominent granular cytoplasmic stippling. Immunohistochemically, tumor cells in all cases were immunoreactive for CK20. Signal pathway related protein mTOR and S6 were positive in 14/15 and 6/15 cases, respectively. Cathepsin K, Melan A and HMB45 were at least focally positive in 12/15, 6/15 and 2/15 cases, respectively. CK7 and CD10 showed focal immunostain positivity in some cases, while TFE3, TFEB, CA9 and CD117 were negative in all cases. NGS demonstrated TSC1/TSC2 mutations in all tested cases (10/10). Conclusions ESC RCC is a rare tumor that tends to occur in young patients with an indolent behavior. Diagnosis can be established by its distinct clinical and histopathologic findings, immunohistochemical phenotype and molecular genetics. The tumor may be considered as a new subtype of RCC.

Objective: To study the clinicopathologic features, immunophenotype, characteristic FISH pattern and prognosis of renal cell carcinoma (RCC) associated with chromosome X inversion harboring gene fusions involving TFE3. Methods: Ten cases of NONO-TFE3 RCC and four cases of RBM10-TFE3 RCC were investigated at Nanjing Jinling Hospital from 2009 to 2016 by clinicopathological findings, immunohistochemistry, and genetic analysis. Results: Morphologically, the distinct pattern of secretory endometrioid subnuclear vacuolization was overlapped with clear cell papillary RCC, and often accompanied by sheets of epithelial cells in NONO-TFE3 RCC. Most cases of RBM10-TFE3 RCC presented with the biphasic feature that acinar, tubular and papillary patterns of epithelioid cells combined with sheets of small cells with "pseudorosette-like" architectures. In addition, cytoplasmic vacuolization, nuclear groove, and psammoma bodies were also observed. Immunohistochemically, all NONO-TFE3 RCC cases were immunoreactive for TFE3, CD10, RCC markers, and PAX8, and negative for CK7, Cathepsin K, Melan A, HMB45, Ksp-cadherin, vimentin, and CD117. All 4 cases of RBM10-TFE3 RCC showed moderate to strong immunoreactivity for TFE3, Cathepsin K, CD10, Ksp-cadherin, E-cadherin, P504s, RCC marker, PAX8, and vimentin but negative for TFEB, HMB45 and CK7. CKpan and Melan A were at least focally expressed. The antibody to Ki-67 showed labeling of 3%-8% (mean 5%). There were some expression discrepancies of immunochemistry between different histological patterns. PAX8, CKpan, P504s, and Ksp-cadherin were expressed in epithelioid areas but not in small-cell areas. Ki-67 labeling index of epithelioid areas was higher than that in small-cell areas. In molecular analysis, NONO-TFE3 fusion transcripts were identified in 6 patients. The fusion points were between exon 7 of NONO and exon 6 of TFE3 in 5 patients and between exon 9 of NONO and exon 5 of TFE3 in one patient. All 4 cases of RBM10-TFE3 RCC demonstrated to have RBM10-TFE3 fusion transcripts and the fusion points were between exon 5 of TFE3 and exon 17 of RBM10. Using TFE3 break-apart FISH assay, all 10 cases of NONO-TFE3 RCC showed characteristic patterns of equivocal split signals with a distance of nearly 2 signal diameters. All 4 cases of RBM10-TFE3 RCC showed colocalized or subtle split signals with a distance of <1 signal diameter, which was considered as negative results. Long-term follow-up was available for 7 patients of NONO-TFE3 RCC and 4 patients of RBM10-TFE3 RCC. All patients were alive with no evidence of disease. Conclusions Two rare genotypes, NONO-TFE3 RCC and RBM10-TFE3 RCC, are reported in this study. Both of these two tumors show specific morphology and good prognosis, along with the positive TFE3 staining and the equivocal or false-negative TFE3 FISH results, which could be missed. PCR detection or next-generation sequencing can determine the genotype.