Objective To explore the diagnosis and surgical treatment of primary intestinal tumors (PIT), to improve the diagnosis and treatment. Methods Retrospectively analysis was made on the clinical data of 72 patients with PITs admitted to our hospital from 1988 to 2002. Results Out of the 72 cases, 20.8% (15/72) had benign tumors, while the remaining 79.2% (57/72) were malignancies. The former were mostly adenoma and liomyoma, each accounting for 40.0% (6/15) of the benign tumors. Adenocarcinoma was the most common type of malignancy (36.8%,21/57), following lymphadenoma (30.0%, 17/57). The main diagnostic methods were X-ray,B-ultrasonic,CT, endoscopy and superior mesenteric arteriography.The misdingnosis rate was 62.5% before operation in this series.Of the 72 cases underwent operation,25 underwent emergent operation (33.3%, 25/72), because acute intestinal obstruction, digestive tract bleeding or perforation,acute appendicitis were diagnosed before the operation.In this series,there was no operative death;the 1,3,5 year survial rates of malignance were 62.5%,47.5%,25.0%,respectively. Conclusions PIT is not easily diagnosed before operation, and the misdiagnosis rate and emergent operation rate are high. Superior mesenteric arteriography,radiologic contrast examination of the small bowel are the important means of diagnosis in jejunum or ileum tumour, and the best way to diagnosis duodenal neoplasms is hypotonic duodenography and endoscopy. Once the diagnosis of PIT is made, the best choice of treatment is operation.

Objective To explore the effect of CYP3A5 gene polymorphisms on the whole blood trough concentration (C0) of tacrolimus (TAC) in patients with idiopathic membranous nephropathy to identify an economical and optimal initial dosage delivering the best curative effect with minimum drug adverse reaction.Methods Sixty patients with idiopathic membranous nephropathy were enrolled in this study.The CYP3A5 genotype was tested by fluorescence in situ hybridization (FISH).According to CYP3A5 genotype, the patients were divided into three groups (AA, AG, and GG).At the same time, the C0 of TAC was measured by enzyme multiplied immunoassay technique (EMIT).C0 of TAC, daily dosage of TAC and the concentration/dose(C0/D) ratio of TAC were detected after taking medicine at 8, 12, 16 and 24 weeks respectively, so as to corroborate the relation between CYP3A5 gene polymorphisms and the dosage of TAC.Results The oral TAC dosage had great variation among individuals.The occurrence of the CYP3A5 genetic polymorphisms (A6986G) designated as G was 53.33％.D and C0 were significantly different at 8, 12, 16 and 24 weeks respectively (all P ＜ 0.05).To reach the same C0, the patients with AA needed 2-3-fold dosage of TAC than GG;and those with AG needed 1-2-fold dosage of TAC than GG.After 24-week treatment, the effective rate of AA group was markedly lower than AG and GG (16.67％ vs 81.25％, 16.67％ vs 87.50％, all P ＜ 0.001).Among CR, PR and NR, there were no significantly difference on C0 or C0/D of TAC (P ＞ 0.05).Conclusions CYP3A5 genotypes are correlated with blood concentration of TAC.CYP3A5 genotyping may be a new approach to predict the optimal initial dosage of tacrolimus in idiopathic membranous nephropathy.

Objective To study the effect of astragaloside on TGF-β1,SMAD2/3,and α-SMA expression in the kidney tissue of diabetic KKAy mice,and evaluate its potential role in renal interstitial fibrosis.Methods 20 type 2 diabetic KKAy mice were randomly divided into model group and astragaloside group,while 10 male C57BL/6J mice were selected as the control.Astragaloside at 40 mg · kg-1 · d-1 was given when the KKAy mice fed with high-fat diet to 14 weeks old.The mice in the control and model group received normal saline at 40 mg · kg-1 · d-1.Blood glucose meter was used to detect the blood glucose value of each mice at 16th,20th and 24th week.The mice were killed at 24 weeks old and the kidney tissue samples were collected.Pathology morphological changes were observed.Results (1) blood glucose value:cmpared with the control group,the blood glucose value of KKAy mice at 14 week increased significantly,and that of model group also increased significantly at 16th,20th and 24th week (P＜0.05);the blood glucose value of astragaloside group decreased compared with control group (P＜0.05).(2) Morphology of kidney:in the control group,the glomerular and tubular had clear structure,there was no renal interstitial fibrosis;in the model group,the renal glomerular mesangial matrix had broaden,mesangial cell had increased,renal tubular epithelial cell cytoplasm showed vacuole degeneration,renal interstitial inflammatory cell had increaised.In astragaloside group,there were few renal tubular epithelial cell cytoplasm,and there was no obvious fibrosis.(3)TGF-β1,SMAD2/3,and α-SMA expression levels of the kidney issuse:compared with control group,mice in model group up-regulated TGF-β1,SMAD2/3 and α-SMA expression (P＜ 0.05).TGF-β1,SMAD2/3,and α-SMA expression levels in astragaloside group were significantly lower than those in the model group (P＜0.05).There was few phosphorylated SMAD2/3 expression in renal tubular and glomerular nuclei,while that of model group increased (P＜0.01),and compared with model group,that of the astragaloside group decreased (P＜0.05).Conclusion Astragaloside can delay the renal fibrosis process in diabetic mice by influencing the TGF-β/SMADS signaling pathway and down-regulating TGF-β1 and α-SMA expression,thus to relieve renal fibrosis in diabetic mice.

ISG15, the first ubiquitin-like molecule identified two decades ago, is encoded by interferon stimulated gene 15 ( ISG 15), where its robust expression can be induced by viral infections or interferon treatments. ISG 15 conjugate to other proteins as the ubiquitin and was found to be involved in innate immune response. However, the functions of ISG15 modification remained unclear. We cloned bovine ISG15(bisg15) into a prokaryotic expression vector pET28a( + ) with a His-tag to generate a soluble form of bISG15 fusion protein, and purified with Ni-NTA Sepharose chromatography. The purified protein was concentrated and used to immune Balb/c mice to raise the antiserum, which could specifically recognize bISG15 expressed in eukaryotic cells by Western blot analysis. The concentrated bISG15 protein and its antiserum were then used to establish an in vitro bISG15 modification system. Our studies have demonstrated that cellular proteins could be conjugated to bISG15 with this system.

The ubiquitin-like modifier bISG15 is an antiviral protein found in fetal bovine lung (FBL) cells.Bovine Herpesvirus 1(BHV-1),which is a viral pathogen of cattle,can infect FBL cells and induce cytopathic effects.Real-time PCR assays showed that BHV- 1 's infection could repress the basal or inducible transcription of bISG15 in FBL cells.It demonstrates that this repression effect depends on BHV-1 viral infection and new protein synthesis.Our previous work showed that bIRF-3 was the key factor in the stimulation of bISG 15 in FBL cells,so the effect of BHV-1 viral protein on bIRF-3 activating the promoter of bISG15 was confirmed.The luciferase assay showed the BHV-1 viral protein bICP0 inhibited the activation of bISG15 promoter stimulated by bIRF-3.Taken together,our work suggested that BHV-I had some molecular mechanism to resist the cellular bISG15'santiviral functions.

Objective Endogenous cadiac stem cells become the famous star in the cardiac regeneration field in recent years.But the biological behavior of cardiac stem cells has not been fully explained clearly.This experiment intends to research the cardiac stem cell niche and its characteristics,as well as the succinct method for cardiac stem cells (Cardiospheres derived cells) culture in vitro.Methods Hearts from four 8-week-old SD rats were cut into pieces less than I mm3.Then,the small tissues were digested by trypsogen and collagenase in turn.And the undigested myocardium was collected and cultured in media until the de novo cells bespread the pertri-dish bottom.The 3rd generation of cardiac stem cells were harvested and prepared for immunofluorescence to detect the expression of related molecular markers,so as to identify the differentiation of these stem cells.Then,the cultural tissues were collected and embedded in paraffin,and the H-E staining,Masson staining,immunoflnorescence and TUNEL apoptosis assay were carried out.In addition,EdU was added into the medium and cuhured for 72 hours before the cultural tissues were harvested so as to label the proliferative cells.Results After 7 ～ 10 days in culture,some small,round and phase-bright cells aresed from the adherent plated tissue.These cells had the spontaneous differentiation potency in vitro culture.The immunofluorescence shows mitotic phase cells were c-kit positive,and most of passage cells were α-sarcomeric actin 、cardiac troponin T、flk-1 and vemintin positive while CD90 negative,but few of them were α-smooth muscle actin 、conexin-43 、CD31 and CD45 positive.There were lots of newborn cells grow exuberantly while accompanied apoptosis of myocardial cells in the paraffin section.The myocardial collagen remodels in the cultural myocardial tissues at the same time.The proliferative cells in the cultural tissues were EdU positive.Throe myocardium within the cultural tissues were MMP-2、MMP-9 and TGF-β1 positive,On the contrary,the proliferative cells were almost negative when stained with those antibodies.Conclusion Conclusions It's a convenient way to harvest cardiac stem cells by the use of myocarlial? tissues cultured in vitro.Those stem cells grow and proliferate in the cultural myocardial tissues accompanied with the degradation of extracellular matrix.Cardiospheres derived stem cells have the spontaneous differentiation potency when cultured in vito,and should be a promising? seed cells for stem cell therapy.

Objective:To study clinical features of rheumatoid arthritis (RA) patients with peripheral neuropathy (PN).Methods:The clinical data of 46 RA patients with PN in the First Affiliated Hospital of Zhengzhou University from August 2012 to August 2019 were retrospectively analyzed, including clinical manifestations, laboratory and imaging results, previous treatment, treatment and clinical outcome. The other 92 RA patients without PN at the same period were selected as controls.Results:In RA patients with PN, the male to female ratio was 1∶2.1 with an average age (59.1±11.8) years. The course of RA and PN was 102.0 (19.0-156.0) months and 4.2 (0.7-5.5) months respectively. Numbness (84.8%, 39/46) and muscle weakness (21.7%, 10/46) were the most common symptoms. According to results of electromyography, polyneuropathy (60.0%, 27/46) was the predominant manifestation, followed by mononeuritis multiplex (31.1%, 14/46). Compared to RA patients, rheumatoid factor (RF) ( P<0.001) and the percentage of cutaneous vasculitis ( P=0.042) were higher in RA patients with PN. Logistic regression analysis revealed significant correlation between RF>178.4 IU/ml ( OR＝5.626, 95% CI 2.509-12.618, P<0.001) and development of PN. Paresthesia in 27 patients (58.7%, 27/46) were relieved after treatment of high dose glucocorticoid and immunoglobulins (IVIG). Twelve patients were followed up regularly and the mean duration of follow-up was 17.0(4.8-52.8)months. Paresthesia in 10 (10/12) patients were relieved compared to that at discharge, 1 (1/12) patient achieved complete remission. Conclusion:Numbness and muscle weakness are the common symptoms in RA patients with PN and polyneuropathy is the main type. RF>178.4 IU/ml is correlated with the development of PN in RA patients. Intensive treatment such as high dose glucocorticoid and IVIG are effective.

Virus infection or interferon can stimulate robust expression of the protein ISG15 that is encoded by interferon stimulated gene 15, which was the first unbiquitin-like molecule identified two decades ago. While ubiquitin and its many important functions have been well established, the functions of ISG15 and its post-translational conjugation are still largely unknown . Recently, some specific enzymes have been identified to be involved in the ISG15 modification system, suggests that ISG15 and its modification system play important roles in the innate immune response and regulation of interferon signaling. The history of ISG15 discovery and its biochemical characterization were briefly introduced. Then such topics as the ISG15 gene expression and the ISG15 modification will be focued on, and finally summarize new findings which have implications for ISG15 and its modification system in immunology and interferon signal transduction were summarized.

Objective:Through detecting miRNA-26a,β-catenin,GSK-3β and α-SMA expressions in IgA nephropathy with varying degrees of renal interstitial fibrosis,the study was performed to explore the effect of miRNA-26a targeting GSK-3β on Wnt/β-catenin signal pathway simulated renal interstitial fibrosis.Methods:Incorporated 46 cases of IgA nephropathy patients were divided into three group based on the degree of renal interstitial fibrosis,namely,mild group,moderate group and severe group;7 cases of normal renal tissues away from the renal tumor tissues were selected as the control group.Expression levels of miRNA-26a in renal tissues of each group were detected based on RT-qPCR method,to analyze the correlation between miRNA-26a and renal fibrosis In patients with IgA nephropathy.Furthermore,mRNA and protein expression levels of β-catenin,GSK-3β and α-SMA in renal tissues were measured using RT-qPCR and immunohistochemistry,respectively,the comparison was then made in each group;subsequently,correlation analysis was further conducted to investigate the relationship of miRNA-26a with β-catenin,GSK-3β and α-SMA.Results:(1) Compared with the control group,miRNA-26a expression was down-regulated from renal biopsy of IgA nephropathy patients,the expression level of miRNA-26a was significantly decreased,showing statistical differences among groups (P＜0.05).(2) Compared with the control group,mRNA and protein expression levels of β-catenin,GSK-3β and α-SMA in renal tissues were all increased in IgA nephropathy patients,and the degree of expression increased gradually with the increase of the degree of renal interstitial lesion,differences were statistically significant among groups (P＜0.05).(3) Correlation analysis results indicated that there were negative correlation between miRNA-26a expression in renal tissues and the degree of renal interstitial fibrosis,differences were statistically significant among groups (r=-0.943,P ＜0.05),at the same time,expression intensities of GSK-3β,β-catenin and α-SMA in renal interstitium and renal tubules were positively correlated with the degree of renal interstitial fibrosis(r =0.917,P＜0.05;r =0.943,P＜0.05;r =0.926,P ＜0.05),meanwhile,positive correlation was also found regarding protein expression of GSK-3β and β-catenin in renal interstitium (r=0.834,P＜0.05).Conclusion:Collectively,miRNA-26a can be involved in Wnt/β-catenin signal pathway simulated renal interstitial fibrosis via the regulation of GSK-3β.

Besides its function as a pathogenicity determinant, the Tombusvirus P19 also serves as a suppressor of RNA interference (RNAi) by sequestering intracellular small RNAs such as the small interfering RNAs (siRNAs) and microRNAs (miRNAs). However, the effect of P19 on mammalian cells has not been evaluated before. A human embryonic kidney 293 cell line that stably expressed p19 (HEK293-p19) was generated. Flow cytometric analysis revealed that over-expression of P19 caused a significant accumulation of G2/M phase cells. Cell proliferation assays demonstrated a reduced DNA replication and cell growth in HEK293-p19 cells. Moreover, p19 altered the expression profiles of a number of cell cycle regulators in HEK293 cells, such as upregulafion of cyclin A1, CDK2, CDK4, CDK6, p18, cyclin D2, p19INK4d and E2F1, and downregulation of p15, cyclin A2, cyclin B1 and cyclin E1. Thus, the data strongly indicate that p19 might influence multiple G2/M regulators to cause G2/M arrest.