Altered brain iron deposition in patients with minimal hepatic encephalopathy: an MRI quantitative susceptibility mapping study.

AIM: To explore the use of quantitative susceptibility mapping (QSM) in assessing changes in brain iron deposits and their association with cognitive function in patients with minimal hepatic encephalopathy (MHE). MATERIALS AND METHODS: The study cohort comprised 27 cases with hepatitis B-associated cirrhosis with MHE (MHE group), 25 with hepatitis B-associated cirrhosis without MHE (NMHE group), and 25 healthy controls (HC group). Iron deposits in the bilateral frontal white matter, caudate nucleus (CN), putamen, globus pallidus, thalamus, red nucleus, substantia nigra (SN), hippocampus, and dentate nucleus were measured by QSM. The associations between iron deposition with the time taken to complete number connection tests A (NCT-A) and the score on digital-symbol test (DST) were analysed. RESULTS: Susceptibility values differed significantly in the bilateral CN, left thalamus, right SN, and left hippocampus in the MHE group compared with the other groups and were positively associated with the times taken to complete the NCT-A in the bilateral CN, left thalamus, and right SN and negatively associated with DST scores in the bilateral CN, left TH, and left HP. CONCLUSION: Reduced cognitive function in MHE patients was significantly associated with abnormally increased iron deposition in certain brain areas. The quantification of brain iron deposition by QSM may thus be an objective and accurate means of evaluating MHE.

[Pulmonary nuclear protein of the testis midline carcinoma:a case report].

Nuclear protein of the testis (NUT) midline carcinoma (NMC) is a rare malignant epithelial tumor that typically occurs in the midline regions such as the head, neck, and mediastinum. This tumor is characterized by rapid development, aggressive growth, and strong invasiveness. Due to its short duration, most patients are diagnosed at advanced stages, often leading to rapid mortality. Although reports on pulmonary NUT carcinoma are uncommon, this article presents a case of pulmonary NUT carcinoma in which the patient repeatedly expectorated bronchial casts and tumor tissue. Additionally, a comprehensive review of relevant literature from recent years is provided to enhance understanding of this disease.

[Preliminary exploration of modified side overlap with fundoplication by Yamashita (mSOFY) anastomosis technique in laparoscopic proximal gastrectomy].

Objective: To discuss the feasibility and safety of modified side overlap with fundoplication by Yamashita (mSOFY) in laparoscopic proximal gastrectomy. Methods: Using the method of descriptive case series study, the clinical data of 9 patients with upper gastric cancer who successfully performed mSOFY anastomosis from March 2022 to October 2022 in the Affiliated Huaian No.1 People's Hospital of Nanjing Medical University were retrospectively analyzed.The reconstruction steps of mSOFY anastomosis are as follows: (1) Make a small incision on the right side of the esophageal stump and in front of the anterior wall of the gastric stump; (2) The 45mm linear cutting stapler is placed into the preset anastomosis of the esophagus and the remnant stomach, and the esophagus is rotated 90° counterclockwise along the axis, so that the right wall of the esophagus is anastomosed with the remnant stomach, and the stomach wall is sutured to the left side of the esophagus; (3) The common opening of esophagus and remnant stomach was sutured with inverted suture; (4)Suture the left and lower sides of the esophagus with the remnant stomach to make the esophagus flat against the stomach wall; (5) Open the sutured common opening: due to the pressure of the false dome, the posterior wall of the lower esophageal segment was compressed into a valve-like structure. We mainly observing the postoperative reflux and nutritional improvement of the patients, and recording the intraoperative situation and postoperative complications. Results: Nine patients with upper gastric cancer who completed laparoscopic proximal gastrectomy (mSOFY anastomosis) did not have conversion to laparotomy or intraoperative / postoperative complications. The operation time was (169.4±10.4) minutes, the anastomotic reconstruction time was (51.7±7.1) minutes, the intraoperative bleeding volume was (98.9±43.4) ml, and the number of lymph nodes dissected was (27.2±6.7). The patient recovered well after operation, without any complaints related to reflux esophagitis. Postoperative gastrointestinal radiography showed that the anastomosis was smooth, without stenosis and leakage. The serum albumin [(41.6±3.4) L vs. (39.9±2.6) L], prealbumin [(211.3±38.6) mg/L vs. (205.3±36.0) mg/L], and hemoglobin levels [(126.7±13.2) g/L vs. (121.0±9.7) g/L] of patients before and one month after surgery have no statistically significant differences (all P>0.05). Conclusion: mSOFY anastomosis can be used as one of the safe and feasible reconstruction methods in laparoscopic proximal gastrectomy.

[Clinical efficacy evaluation of minimally invasive cardiac surgery coronary artery bypass grafting in patients with multivessel coronary artery disease].

Objective: To investigate the clinical efficacy of minimally invasive cardiac surgery coronary artery bypass grafting (MICS CABG) in the treatment of patients with multivessel coronary artery disease. Methods: A total of 147 patients with multi-vessel coronary artery disease and coronary heart disease who underwent surgical treatment in Henan Provincial People's Hospital from March 2016 to March 2019 were retrospectively selected. Of which, 69 patients were treated by MICS CABG (minimally invasive group) and 78 patients were treated using the traditional thoracotomy (traditional group). The perioperative indexes, serum myocardial enzyme indexes and renal function indexes of patients before and after operation were compared between the two groups; Two groups of patients were followed up for 2 years; the incidence of adverse cardiovascular events (MACE) was recorded, and survival analysis was performed. Results: The age of the patients in the minimally invasive group and the traditional group were (66.9±5.8) and (68.2±7.0) years old, respectively, and the proportions of males were 60.9% (42 cases) and 51.3% (40 cases) (all P>0.05). All patients in the two groups successfully completed the operation, and no patients in the minimally invasive group were converted to thoracotomy. Before surgery, there was no significant difference in serum cTnI, CK-MB, BUN, Scr, and creatinine clearance between the minimally invasive group and the traditional group (all P>0.05). After re-examination 48 hours after operation, the serum cTnI in the minimally invasive group was (3.109±0.664) µg/L, and the CK-MB was (18.03±3.27) U/L, which were lower than those in the traditional group (3.438±0.715) µg/L, (20.63±4.28) U/L; the difference was statistically significant (all P<0.05). During the 2-year follow-up, there was no statiscally significant difference in the incidence of recurrent myocardial infarction, postoperative atrial fibrillation, postoperative stroke, arrhythmia, heart failure, thrombosis, cardiac death, and MACE events between the minimally invasive group and the traditional group. Statistical significance (all P>0.05). The survival curve analysis showed that the cumulative rates of MACE events in the minimally invasive and traditional groups were 17.39% and 26.92%, respectively (P=0.171). Conclusions: The effect of MICS CABG in the treatment of patients with multivessel coronary artery disease and coronary heart disease is not much different from that of traditional open thoracotomy, but the former is less traumatic, quicker after surgery, and has clinical significance for the recovery of patients' myocardial function.

[The value of preservation of subvalvular structures in patients with mitral and aortic valve replacement surgery and its effect on left ventricular contractility].

Objective: To discuss the clinical value of preserving subvalvular structure in mitral and aortic valve replacement surgery and its effect on left ventricular contractility. Methods: A total of 97 patients who underwent mitral valve replacement surgery in the Adult Cardiac Surgery of Heart Center of Henan Provincial People's Hospital, Central China Fuwai Hospital from June 2016 to December 2018 were selected as the research subjects, of whom 45 cases were preserved subvalvular structure and 52 cases were in the total resection group (intraoperative total resection of the mitral valve and subvalvular chordae tendineae). General cardiac function indexes and left ventricular function quantitative indexes were compared before and in 3 months and 6 months after the operation of the two groups; The changes of the overall longitudinal strain of the long axis of the apex and the overall circumferential strain of the short axis of the left ventricle determined by the two-dimensional speckle tracking technology were compared before and after the operation. Results: The ages of the patients in the preservation group and the total resection group were (41.8±11.3) and (43.3±10.6) years old, respectively, and the male proportions were 58.0% (26 cases) and 44.0% (23 cases), respectively, with no significant difference (all P>0.05). The aortic occlusion time and cardiopulmonary bypass time of the patients in the preservation group were (57.8±4.5) and (78.6±6.7) min, respectively, which were longer than those in the total resection group [(48.1±4.4) and (48.1±4.4) min, respectively] (all P<0.05). The left atrial pressure of the patients in the preservation group at shutdown was (8.4±1.8) mmHg (1 mmHg=0.133 kPa), which was lower than that of the total resection group (11.3±2.5) mmHg (P<0.001). There were interaction effects between groups and time in regards to the left ventricular end-diastolic diameter ( LVEDD ), left ventricular ejection fraction ( LVEF ) and Tei index, as well as the strain rate of mitral annulus and left ventricular wall of interventricular septum of the preservation group and the total resection group (all P<0.05). LVEDD and LVEF of patients in the preservation group at 3rd month after operation were (44.7±4.0) mm and (45.5±4.2) mm, and at 6th months were (56.5±4.9)% and (58.8±5.0)%, respectively, all larger than (42.7±3.6) mm and (42.7±3.6) mm, (54.5±4.6)% and (56.3±4.8)% of the total resection group. The measured value of LVESD in the preservation group at 3rd month after surgery was (32.6±3.2) mm, which was greater than that in the total resection group (31.2±3.4) mm (P<0.05). The Tei index of patients in the preservation group at 3rd and 6th months after surgery were 1.0±0.2 and 0.8±0.2, respectively, which were lower than those in the total resection group 1.2±0.3 and 0.9±0.2 (all P<0.05). Conclusion: Preserving the subvalvular structure during mitral valve replacement surgery can better improve the patient's left ventricular function and left ventricular systolic capacity.

[Diffuse midline gliomas with H3K27 alteration in children: a clinicopathological analysis of forty-one cases].

Objective: To investigate the clinicopathological features of pediatric diffuse midline glioma with H3K27 alteration and to analyze their relationship with prognosis. Methods: Forty-one cases of childhood diffuse midline glioma with H3K27 alteration were collected at Children's Hospital of Fudan University (39 cases) and Xi'an Children's Hospital (2 cases), from July 2016 to July 2020. The clinical manifestations, imaging data, histopathology, immunohistochemical phenotype and molecular genetics features, tumor size, site and histological grading were evaluated. Results: Among the 41 cases, 21 were males and 20 females, the age of onset was 3-14 years, the average and median age was 7.6 years and 7.0 years, respectively. The tumor sites were brain stem (n=36) and other locations (n=5). The clinical manifestations were dizziness, gait disturbance, and limb weakness, etc. The MRI features were variable. The histology varied from low-grade to high-grade glioma with neuron differentiation. Immunohistochemistry showed that the tumor cells expressed H3K27M, GFAP, and Olig2. Genetic study showed that 76% (16/21) of tumors had H3F3A gene mutation, mostly accompanied by TP53 (62%, 13/21) missense mutation; five tumors (24%, 5/21) had HIST1H3B gene mutation, accompanied by missense mutations in ACVR1 and PI3K pathway-related gene PIK3CA (4/5) and PIK3R1 (1/5) mutations. The prognosis was dismal with only one alive and others died. The average and median overall survival time was 7 months and 4 months, respectively. Cox multivariate regression analysis showed that age, tumor location, radiologically maximum tumor diameter, histologic grading, and surgical methods were not significantly associated with overall survival rate (P>0.05). Conclusions: Pediatric diffuse midline gliomas with H3K27 alteration have unique clinicopathological and genetic characteristics. The prognosis is poor. The tumor location and histopathologic grading are not related to prognosis. New specific drugs and comprehensive treatment are needed to improve the prognosis.

Effects of lncRNA MALAT1 and lncRNA NKILA on proliferation, invasion and apoptosis of retinoblastoma.

OBJECTIVE: To assess the expression of LncRNA NKILA and MALAT1 in retinoblastoma and its related mechanisms. PATIENTS AND METHODS: Tixty-eight cases of retinoblastoma patients admitted to our hospital from February 2017 to February 2019 were collected as a research group, while 70 healthy people who came to our hospital for checkup at the same time were chosen as a control group. Both retinoblastoma and human colorectal mucosa cells were purchased, expression and clinical value of NKILA and MALAT1 in serum of Rb patients were tested, and sh-NKILA, si-NKILA, NC, sh-MALAT1 and si-MALAT1 were transfected into Weri-Rb1 and Y79 cells. qRT-PCR was adopted to detect the NKILA and MALAT1 levels in samples, and WB was adopted to detect the cle-caspase-3, cle-caspase-9, Bax, Cyclin B1, CDC2 and p-CDC2 protein levels in cells. Cell proliferation was conducted via MTT assay, invasion was carried out through transwell assay, and apoptosis was confirmed by flow cytometry assay. RESULTS: NKILA and MALAT1 were low expressed in retinoblastoma, and AUC of LncRNA NKILA and MALAT1 was over 0.8. LncRNA NKILA and MALAT1 were associated with tumor size, classification and clinical grading in children with retinoblastoma. Over-expression of NKILA and MALAT1 could promote apoptosis, inhibit cell growth and Bcl-2 protein, and promote upregulation of the expression levels of clecaspase-3, clecaspase-9 and Bax. CONCLUSIONS: By regulating MALAT1 and NKILA, we controlled the growth and apoptosis of Rb cells, which was expected to be a potential clinical therapeutic target for Rb.

MiR-181a affects myocardial ischemia-reperfusion injury in rats via regulating akt signaling pathway.

OBJECTIVE: The aim of this study was to explore the influence of the micro ribonucleic acid (miR)-181a on myocardial ischemia-reperfusion injury (MIRI) in rats by regulating the protein kinase B (Akt) signaling pathway. MATERIALS AND METHODS: A total of 30 male Sprague-Dawley rats were randomly divided into three groups, including: sham operation group (Sham group), ischemia-reperfusion group (I/R group), and miR group (MiR-181a group). The model of myocardial ischemia-reperfusion was successfully established in rats. The concentration of blood nitric oxide (NO) was detected by the relative kits. Myocardial apoptosis in rats of the three groups was detected using terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay. Furthermore, the expressions of myocardial cell apoptosis-related proteins and tumor necrosis factor-α (TNF-α), and the degree of Akt phosphorylation were determined by Western blotting. RESULTS: Compared with Sham group and miR-181a group, I/R group exhibited significantly elevated left ventricular end-diastolic pressure (LVEDP) (p<0.05). However, the left ventricular end-systolic pressure (LVESP), stroke work (SW), differential pressure (DP), end-systolic pressure-volume relationship (ESPVR), and end-diastolic pressure-volume relationship (EDPVR) significantly decreased in the I/R group (p<0.05). In comparison with miR-181a group, the apoptosis index of myocardial cells was remarkably elevated in the I/R group, showing statistically significant differences (p<0.05). The protein bands were analyzed using the Quantity One detection software. The results demonstrated that, compared with the Sham group, I/R group showed significantly elevated expressions of cysteine-aspartic protease (Caspase)-3 and TNF-α in rat myocardial tissues (p<0.05). However, the protein levels of Akt and endothelial NO synthase (eNOS) phosphorylation and NO in rat myocardial cells were significantly down-regulated (p<0.05). CONCLUSIONS: MiR-181a activates Akt to promote the phosphorylation of its downstream protein eNOS, inhibit the apoptosis of myocardial cells, and alleviate MIRI.

MiR-34a affects hepatocyte proliferation during hepatocyte regeneration through regulating Notch/HIF-1α signaling pathway.

OBJECTIVE: To explore the influences of micro ribonucleic acid (miR)-34a on liver function and hepatocyte proliferation during hepatocyte regeneration in rats and its mechanism. MATERIALS AND METHODS: A total of 80 Sprague-Dawley rats were randomly divided into 4 groups: Sham-2 d group (2 days after hepatectomy), Sham-10 d group (10 days after hepatectomy), miR-34a siRNA-2d group (miR-34a knockdown + 2 days after hepatectomy) and miR-34a siRNA-10 d group (miR-34a knockdown + 10 days after hepatectomy), with 20 rats in each group. Serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), and lactate dehydrogenase (LDH) were detected at 2 d and 10 d after the operation. The rat liver was harvested for calculating the liver/body weight ratio. In addition, the deoxyribonucleic acid (DNA) content in rat hepatocytes was detected via Feulgen staining. The pathological changes in rat liver were detected via hematoxylin-eosin (H&E) staining. Moreover, the hepatocyte apoptosis in each group was detected via terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining. Expression levels of proliferating cell nuclear antigen (PCNA), Notch1 intracellular domain (NICD), and hypoxia-inducible factor-1α (HIF-1α) in liver tissues of each group were detected via immunohistochemistry and Western blotting. RESULTS: No significant differences in the liver/body weight ratio, serum levels of ALT, AST, LDH, pathological structure of the liver, hepatocyte apoptosis level, and PCNA expression in hepatocytes were found between miR-34a siRNA-2 d group and Sham-2 d group. However, the expression levels of NICD and HIF-1α in the liver significantly increased in miR-34a siRNA-2 d group compared with those in Sham-2 d group (p<0.05). On the contrary, compared with those in Sham-10 d group, the liver function and hepatocyte regeneration level significantly increased in miR-34a siRNA-10 d group. Increased liver/body weight ratio, remarkable decline in serum levels of ALT, AST, and LDH, significant alleviation of pathological injury of liver tissues, decreased the apoptosis level and upregulated PCNA protein were observed in miR-34a siRNA-10 d group than those of Sham-10 d group. The Notch/HIF-1α signaling pathway was also significantly activated. CONCLUSIONS: MiR-34a knockdown can significantly enhance the liver function and hepatocyte regeneration ability in rats at 10 d after hepatectomy through activating the Notch/HIF-1α signaling pathway.

Therapeutic effects of probiotic Clostridium butyricum WZ001 on bacterial vaginosis in mice.

AIMS: To observe the therapeutic effects of vaginal infusion of probiotic Clostridium butyricum WZ001 on bacterial vaginosis (BV) in mice. METHODS AND RESULTS: Female ICR mice were used to establish the model of BV by infecting oestrogen-treated mice with Escherichia coli, and then treated with high- and low dose of C. butyricum. Clinical indexes of mice in the C. butyricum-treated groups were significantly improved and comparable to those in the antibiotic group. Pap staining showed that neutrophil count was significantly increased after modelling and largely decreased after C. butyricum treatment (P < 0·01). Dynamic observation of E. coli and Lactobacillus showed that the number of E. coli significantly decreased in the C. butyricum-treated groups or in the antibiotic group with prolonged treatment (P < 0·01). Besides, the number of E. coli in the low-dose C. butyricum group was higher than that in either its high-dose counterpart or the antibiotic group respectively (P < 0·01). The number of Lactobacillus decreased evidently in the antibiotic group (P < 0·01), while that in the C. butyricum groups remained consistent. Moreover, C. butyricum inhibited the proliferation of E. coli by the experiment in vitro. The phosphorylation of nuclear factor-kappa B (NF-κB) p65 in vaginal tissue and the serum levels of inflammatory cytokines, IL-1ß, TNF-α and IL-6, increased after modelling and significantly decreased after treated with C. butyricum (P < 0·01), with no difference found when compared with the antibiotic group. CONCLUSION: Clostridium butyricum inhibits the growth of pathogenic bacteria as well as the inflammatory response induced by E. coli and promotes the growth of Lactobacillus to maintain the vaginal micro-ecological balance. SIGNIFICANCE AND IMPACT OF THE STUDY: Our results suggest that probiobitc C. butyricum WZ001 has a great potential in the clinical treatment of BV.

MiR-21 regulates pulmonary hypertension in rats via TGF-ß1/Smad2 signaling pathway.

OBJECTIVE: To investigate the effects of micro ribonucleic acid (miR)-21 on pulmonary hypertension (PH) in rats via regulating tumor growth factor-ß1 (TGF-ß1)/mothers against decapentaplegic homolog 2 (Smad2) signaling pathway and the possible underlying mechanism. MATERIALS AND METHODS: MiR-21 inhibition vector (pLKO-anti-miR-21) was first constructed. The rat model of PH was established by hypoxia feeding induction. A total of three groups were established, including: blank control group, model group and miR-21 low-expression group were set up, with 12 rats in each group. The expression level of miR-21 in lung tissues of rats in each group was detected via quantitative polymerase chain reaction (qPCR). The right ventricle systolic pressure (RVSP) and right ventricular hypertrophy index (RVHI) of rats in each group were measured. The pathological changes in lung tissues of rats were detected using hematoxylin and eosin (H&E) staining. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining was used to detect the level of apoptosis in lung tissues of rats in each group. Furthermore, Western blotting was adopted to detect the expression levels of TGF-ß1/Smad2 signal pathway-related proteins and apoptosis-related proteins in lung tissues of rats in each group. RESULTS: Compared with blank control group, the expression level of miR-21 in lung tissues of rats in model group was significantly increased (p<0.01). Meanwhile, miR-21 expression in lung tissues of rats in miR-21 low-expression group was significantly decreased by transfection of miR-21 inhibition vector (p<0.01). The RVSP and RVHI of rats in model group were significantly higher than those of blank control group and miR-21 low-expression group (p<0.01). H&E staining results indicated that the degree of lung tissue injury in model group was remarkably higher than blank control group and miR-21 low-expression group (p<0.01). According to TUNEL staining results, the number of apoptotic cells in lung tissues of rats in model group was markedly smaller than that of miR-21 low-expression group (p<0.01). Moreover, the expression level of Caspase 3 in lung tissues of rats in model group was significantly lower than that of miR-21 low-expression group, while the expression level of B-cell lymphoma 2 (Bcl-2)/Bcl-2-associated X protein (BAX) was markedly higher. The expression levels of TGF-ß1 and phosphorylated (p)-Smad2 in lung tissues of rats in model group were evidently higher than those of blank control group (p<0.01). In addition, lowly expressed miR-21 could effectively reduce the expressions of TGF-ß1 and p-Smad2 (p<0.01). CONCLUSIONS: MiR-21 regulates the symptoms of PH in rats by activating TGF-ß1/Smad2 signaling pathway.

[Liver transplantation for polycystic liver disease: 11 cases report and literature review].

Objective: To explore the indications and safety of orthopedic liver transplantation for polycystic liver disease (PLD). Methods: Data of 11 patients with PLD who underwent orthotopic liver transplantation between 2004 and 2013 was retrospectively analyzed. Demographic, clinical and follow-up data were collected for statistical analysis. The survival rate was calculated by Kaplan-Meier method. Results: Over a period of 10 years, the patients received modified piggyback orthopedic liver transplantation (n=9) or combined liver-kidney transplantation (n=2) for PLD. The recipients' median age was 56 years. Seven patients were classified as Gigot type Ⅱ PLD, and four were classified as Gigot type Ⅲ PLD. Eight patients had severe decreased mobility (Eastern Cooperative Oncology Group, ECOG≥3). Only three cases were Child-Pguh Class C patients and the model for end-stage liver disease (MELD) score was>20. The mean hospitalization duration was (45.4±15.3) days, and the mean length of stay in intensive care unit was (4.1±1.9) days. The perioperative mortality was 18.2% and morbidity of complications was 63.6%. The median follow-up period was 111 months. Two patients died of severe complications after combined liver-kidney transplantation. One patient died of ischemia cholangitis during follow-up. The actuarial 1-, 5-and 10-year survival rate during the follow-up period was 82.2%, 81.8%, and 65.5%, respectively. Conclusions: Liver transplantation is the only curative and safe procedure for PLD, and it provides a good long-term prognosis and high quality of life for PLD patients. Liver transplantation could be a primary option in treating progressive or advanced PLD.

[Quantitative evaluation of intravoxel incoherent motion diffusion-weighted imaging and three-dimensional arterial spin labeling in Ki-67 labeling index and grading of brain gliomas].

Objective: To investigate the value of multiple parameters derived from intravoxel incoherent motion diffusion-weighted imaging (IVIM-DWI) and three-dimensional arterial spin labeling (3D-ASL) in Ki-67 labeling index (Ki-67 LI) and grading of human brain gliomas. Methods: From December 2015 to May 2018, 45 patients of gliomas confirmed by surgical pathology in Li Huili Hospital, Ningbo Medical Center were divided into low-grade group (20 cases of WHO grade Ⅱ) and high-grade group (12 cases of WHO grade Ⅲ, 13 cases of WHO grade Ⅳ), and the Ki-67 LI of glioma was obtained by immunohistochemistry. All patients, 24 males and 21 females, aged 25-83 years, mean(53±12)years, underwent conventional magnetic resonance imaging (MRI), IVIM-DWI and 3D-ASL before operation, then measured the true water diffusion coefficient (D), microcirculation perfusion coefficient (D(*)), perfusion fraction (f) and cerebral blood flow (CBF) in the tumor solid area and the contralateral normal white matter area. Those parameters and the Ki-67 LI were compared between the low-and high-grade groups with Mann-Whitney U test. Spearman's correlation was used to analyze the correlation between the quantitative parameters and Ki-67 LI. The ROC curve was used to assess the diagnostic efficacy of parameters in the grading assessment of brain gliomas. Results: The D(0.791×10(-3)mm(2)/s) and f (0.261) of the high-grade group were lower than those of the low-grade group, whereas D(*) (4.153×10(-3) mm(2)/s), CBF(102.027 ml·min(-1)·100 g(-1)) and Ki-67 LI (0.25) were higher (P<0.05). There was a moderate negative correlation between D, f and Ki-67 LI(r=-0.513,-0.457, all P<0.05). There was no significant correlation between D(*) and Ki-67 LI (P=0.571). The area under the curve (AUC) for identifying high-and low-grade gliomas by D, D(*), f and CBF values was 0.965, 0.745, 0.842, and 0.830 respectively (all P<0.05). Conclusion: D and f can be used for quantitative prediction of Ki-67 LI. IVIM-DWI and 3D-ASL are helpful in the grading assessment of gliomas, and the diagnostic efficiency of D is the highest.

Role of miR-16-5p in the proliferation and metastasis of hepatocellular carcinoma.

OBJECTIVE: The aim of this study was to investigate the role of miR-16-5p in hepatocellular carcinoma (HCC), and to explore the possible underlying mechanism. PATIENTS AND METHODS: 100 pairs of cancerous and para-cancerous tissues surgically removed in our hospital were collected. Real Time quantitative-Polymerase Chain Reaction (qRT-PCR) was used to detect the expression level of miR-16-5p in tissues. Bioinformatics and Dual-Luciferase reporter gene assay were used to screen and verify the potential target genes of miR-16-5p, respectively. Human HCC SMMC-7721 cells were used for functional experiments. Cell proliferation was detected by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay. Cell invasion and migration were evaluated by transwell and scratch wound-healing assay, respectively. The protein expression levels of epithelial-mesenchymal transition (EMT) associated markers were measured by Western blot (WB) assay. RESULTS: QRT-PCR showed that miR-16-5p expression in HCC tissues was significantly lower than that of adjacent normal liver tissues. At the cellular level, miR-16-5p was lowly expressed in HCC cells (SMMC-7721). Bioinformatics websites (including Targetscan, PicTar, miRanda) predicted that insulin-like growth factor 1 receptor (IGF1R) was a potential target gene of miR-16-5p. Meanwhile, IGF1R was selected for further investigation due to its metastatic function. The results showed that no significant difference was found in the mRNA expression level of IGF1R in HCC tissues. However, the protein level of IGF1R was significantly up-regulated, which was negatively correlated with miR-16-5p. Combined with Dual-Luciferase reporter gene assay, it was confirmed that miR-16-5p could regulate the expression of IGF1R in a targeted manner. Furthermore, down-regulation of IGF1R significantly reduced the inhibitory effect of miR-16-5p on the proliferation and metastasis of SMMC-7721 cells. CONCLUSIONS: We showed that miR-16-5p suppressed invasion and migration of HCC cells, mechanically by directly targeting and inhibiting IGF1R protein expression. The newly identified miR-16-5p/IGF1R axis might provide new insights into the pathogenesis of HCC and novel potential therapeutic targets for the treatment of HCC.

Postcontrast T1 Mapping for Differential Diagnosis of Recurrence and Radionecrosis after Gamma Knife Radiosurgery for Brain Metastasis.

BACKGROUND AND PURPOSE: The differential diagnosis of radionecrosis and tumor recurrence in brain metastases is challenging. We investigated the diagnostic efficiency of postcontrast T1 mapping in solving this problem. MATERIALS AND METHODS: Between March 2016 and June 2017, fifty-six patients with brain metastases who underwent contrast-enhanced cerebral T1 mapping were recruited for this prospective study. The findings revealed new enhancement after gamma knife radiosurgery. The subjects were assigned to radionecrosis and recurrence groups based on follow-up (median, 11.5 months) and histopathologic results. T1 values of lesions 5 (T15min) and 60 (T160min) minutes after administration of contrast agent and their difference (T1differ) were compared between the 2 groups with the 2-tailed Mann-Whitney U test. Receiver operating characteristic curves were used to determine the optimum cutoff values for differential diagnosis. RESULTS: There were significant differences between the 2 groups in T15min, T160min, and T1differ values (P = .012, P = .004, and P < .001, respectively). Relative to T15min and T160min, T1differ exhibited greater sensitivity and specificity (P < .001, respectively) in identifying radionecrosis. The optimum T1differ value for differential diagnosis was 71.1 ms (area under the curve = 0.97; 95% CI, 0.93-1.00), with sensitivity and specificity of 81.5% and 96.5%, respectively. CONCLUSIONS: Postcontrast T1 mapping is optimal for the differential diagnosis of radionecrosis and tumor recurrence. Among T1 parameters, T1differ is the most powerful parameter for differential diagnosis. Advantages in terms of quantitative analysis and high resolution portend the wide use of postcontrast T1 mapping in the future.

Effects of oxidative stress induced by high dosage of dietary iron ingested on intestinal damage and caecal microbiota in Chinese Yellow broilers.

The objective of this trial was to test the effects of oxidative stress induced by a high dosage of dietary iron on intestinal lesion and the microbiological compositions in caecum in Chinese Yellow broilers. A total of 450 1-day-old male chicks were randomly allotted into three groups. Supplemental iron (0, 700 and 1,400 mg/kg) was added to the basal diet resulting in three treatments containing 245, 908 and 1,651 mg/kg Fe (measured value) in diet respectively. Each treatment consisted of six replicate pens with 25 birds per pen. Jejunal enterocyte ultrastructure was observed by transmission electron microscopy. The results showed that a high dosage of dietary iron induced oxidative stress in broilers. Dilated endoplasmic reticulum (ER), autophagosome formation of jejunal enterocytes and decreased villi were caused by this oxidative stress. Compared to the control, concentration of the malondialdehyde (MDA) in jejunal mucosa in the 908 and 1,651 mg/kg Fe groups increased by 180% (p < .01) and 155% respectively (p < .01); activity of copper-zinc superoxide dismutase (Cu/ZnSOD) increased in jejunum (p < .01); and the concentration of plasma reduced glutathione (GSH) decreased by 34.9% (p < .01) in birds fed 1,651 mg/kg Fe. Gene expression of nuclear factor, erythroid-derived 2-like 2 (Nrf2) and zonula occludens-1 (ZO-1), in the higher dietary Fe groups was enhanced (p < .05). Species of microbial flora in caecum increased caused by oxidative stress. The PCR-DGGE (denaturing gradient gel electrophoresis) dendrograms revealed different microbiota (65% similarity coefficient) between the control and iron-supplemented groups (p < .05). These data suggest high dosage of iron supplement in feed diet can induce oxidative stress in Chinese Yellow broilers, and composition of microbiota in the caecum changed. It implied there should be no addition of excess iron when formulating diets in Chinese Yellow broilers.

RNA interference-mediated silencing of aquaporin (AQP)-5 hinders angiogenesis of colorectal tumor by suppressing the production of vascular endothelial growth factor.

Aquaporin (AQP)-5 is an essential member of AQP family involved in the tumorigenesis of various malignant tumors. However, its role in the angiogenesis of colorectal cancer is unclear and requires further investigation. In this study, a pRNA-H1.1 vector containing the short hairpin RNA (shRNA) targeting AQP5 mRNA was constructed to inhibit the endogenous expression of AQP5 in human umbilical vein endothelial cells (HUVECs). We found that the AQP5-silenced HUVECs acquired decreased proliferation, migration and tube formation ability. AQP5 shRNA also inhibited the enzyme activity of matrix metalloprotease (MMP)-9 in HUVECs without affecting the MMP-2. Further, two colorectal cancer cell lines (HT29 and HCT116) stably transfected with scrambled or AQP5 shRNA were established. The expression and secretion of vascular endothelial growth factor (VEGF)-A (a pro-angiogenic factor) in colorectal cancer cells were downregulated by AQP5 shRNA. HUVECs cultured in low-VEGF conditioned media (CM) obtained from cancer cells developed less vessel-like tubes and had decreased proliferation and migration. The growth and angiogenesis of xenograft tumors were suppressed when the endogenous AQP5 in HT29 cells was knocked down. Tumor samples were additionally collected from patients with colorectal cancer to analyze the expression of AQP5. The immunofluorescence data indicated that AQP5 was expressed in both inner cancer areas and CD31-positive vessels. Taken together, our study suggests AQP5 as a novel anti-angiogenesis target for colorectal cancer.

[Influence of UCHL5 on proliferation and apoptosis of SW527 breast cancer cells].

Objective: To investigate the effect of UCHL5 on proliferation and apoptosis of breast cancer cells. Methods: SW527 cells were infected with lentiviral vector carrying short hairpin RNA to delete the expression of UCHL5. 3-(4, 5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay was used to examine cell proliferation, and subcutaneous transplantation experiments were performed to detect tumor growth. Cell apoptosis was detected using Annexin V/ Propidium iodide (PI) double staining. The correlation between UCHL5 expression and the expressions of proliferation and apoptosis associated genes was analyzed using TCGA breast invasive carcinoma data set. The relationship between UCHL5 expression and breast cancer patients'survival was analyzed using Kaplan-Meier Plotter online tool. Results: After knockdown of UCHL5, A values of SW527 cells on day 2 and day 4 were 0.822±0.017 and 1.045±0.023, respectively, which were significantly lower than 0.976±0.016 and 1.284±0.025 of control cells on day 2 and day 4 (P<0.001). In vivo xenografted mouse model, the volume in UCHL5-suppressed group was (166.90±75.05) mm(3,) significantly smaller than (329.80±35.84) mm(3) in control group (P=0.029). Flow cytometry analysis showed the apoptotic rate of SW527 cells was (8.60±1.13)% after knockdown of UCHL5, significantly higher than (2.95±0.07)% of control group (P=0.020). TCGA database analysis showed that the expression of UCHL5 was positively correlated with the expressions of genes related to cell proliferation, in paralled with the increased expression of UCHL5, the expression of the pro-apoptosis associated genes was decreased. Kaplan-Meier Plotter analysis demonstrated that the overall survival and relapse-free survival of breast cancer patients with high expression of UCHL5 were much shorter (all P<0.001). Conclusions: Down-regulation of UCHL5 inhibits the proliferation and tumor formation and promotes apoptosis of SW527 cells. High expression of UCHL5 may predict poor prognosis of breast cancer patients.

MRI-measured myocardial iron load in patients with severe diabetic heart failure.

AIM: To investigate whether a correlation exists between abnormal myocardial iron status and cardiac lipid deposition as well as other biomarkers in patients with diabetic heart failure (DHF). MATERIALS AND METHODS: Seventeen volunteers (Group 1), 26 patients with non-severe DHF (Group 2), and 25 patients with severe DHF (Group 3) were recruited for this study. Myocardial middle-section T2* mapping and septal 1H magnetic resonance spectroscopy (1H-MRS) were performed using a 3 T magnetic resonance imaging (MRI) machine to assess the iron status and lipid deposition individually. Fasting venous blood was used to examine serum biomarkers. RESULTS: Cardiac T2* (ms) of the three groups were 22.8±2.1, 21.7±1.8, and 18.6±1.3, respectively. The value of Group 3 was significantly lower than that of the other two groups (p<0.001). Myocardial triglyceride (%) levels differed among the three groups (Group 1, 0.53±0.13; Group 2, 1.11±0.29; Group 3, 1.47±0.12; p<0.001). Cardiac T2* was inversely correlated with both cardiac triglycerides and left ventricular ejection fraction (LVEF) in overall participants (Groups 1-3) or Group 3 (each p<0.001). CONCLUSIONS: Abnormal myocardial iron status was found in patients with severe diabetic heart failure. Myocardial lipotoxicity may be responsible for this process.

SPSB3 targets SNAIL for degradation in GSK-3ß phosphorylation-dependent manner and regulates metastasis.

Epithelial-mesenchymal transition (EMT) is a process during which normal epithelial cells acquire mesenchymal characteristics. EMT has a critical role in various human diseases especially in cancer. EMT facilitates tumor initiation and progression by mediating cancer cell stemness and motility. Zinc finger transcription factor SNAIL is one of the most important initiators of EMT. Therefore, it is of great significance to understand the regulating mechanism of SNAIL. In this study, we carried out a luciferase-based genome-wide screening using small interfering RNA library against ~200 of E3 ligases and ubiquitin-related genes and identified SOCS box protein SPSB3 as a novel E3 ligase component that targets SNAIL into polyubiquitination and degradation in response to GSK-3ß phosphorylation of SNAIL. Functionally, we observed that SPSB3 overexpression greatly inhibits tumor metastasis by regulating SNAIL degradation both in vitro and in vivo. The expression of SPSB3 and SNAIL are negatively correlated in human esophageal squamous cell carcinoma tissues, and low SPSB3 expression indicates lymph node metastasis. Moreover, high SPSB3 expression indicates good survivals in various kinds of cancer. Collectively, these findings suggest that SPSB3-mediated SNAIL degradation has a vital role in regulating EMT and cancer progression.