Comparison between traditional and new obesity measurement index for screening metabolic associated fatty liver disease.

Objectives: Obesity measurement indexes have certain screening value for metabolic diseases. To investigate associations between metabolic associated fatty liver disease (MAFLD) and obesity measurement indexes, including traditional indexes (BMI, WC, WHtR) and new indexes (ABSI, BRI, VAI, LAP), and assess their screening ability. Methods: 12,658 subjects aged 18-75 at the Health Center of a Class III Grade A Hospital were included, who were divided into MAFLD and non-MAFLD groups. Spearman's rank correlation was used to study the correlation between MAFLD and obesity measurement indexes. Receiver operating characteristic (ROC) curves were used to calculate the area under the curve (AUC) to evaluate their screening accuracy. Results: MAFLD had strong correlation with traditional BMI and new index LAP. ROC analysis showed that BMI had the highest AUC (0.89), followed by LAP (0.87). Stratification by BMI, LAP had the highest AUC (0.90) for MAFLD in population without obesity (BMI< 23kg/m2), and its optimal cutoff value was 20.75, with a sensitivity and specificity of 85.9% and 79.0%, respectively. Conclusions: We proposed a two-step screening strategy for MAFLD, combining BMI and LAP, and defined a high-risk population for MAFLD as follows: 1) BMI ≥ 23 kg/m2; and 2) BMI< 23 kg/m2 and LAP ≥ 20.75.

MiR-203 is an anti-obese microRNA by targeting apical sodium-dependent bile acid transporter.

Obesity is characterized by excessive fat deposition within the body. Bile acids (BA) are important regulators for controlling the absorption of lipid. Here we show that miR-203 exerts weight-loss and lipid-lowering effects by increasing total BA excretion in obese rodents. miR-203 overexpression transgenic mice are resistant to high-fat diet (HFD)-induced obesity and dyslipidemia. Moreover, the knockdown of miR-203 deteriorates metabolic disorders. ASBT plays important role in regulating BA homeostasis and is a direct target of miR-203. In human intestinal epithelial cells, overexpression of miR-203 decreases the cellular uptake of BA by inhibiting ASBT. Furthermore, TCF7L2 is downregulated in obese mice and acts as a transcription factor of miR-203. The ASBT mRNA level was positively correlated with the body mass index (BMI) of population, while the miR-203 level was negatively associated with BMI. Taken together, these data suggest miR-203 could be a new therapeutic BA regulator for obesity and dyslipidemia.

DDAH2 (-449 G/C) G allele is positively associated with leukoaraiosis in northeastern China: a double-blind, intergroup comparison, case-control study.

Cerebrovascular endothelial dysfunction is involved in the progression of leukoaraiosis. Asymmetric dimethylarginine is a competitive inhibitor of nitric oxide, which is highly expressed in patients with leukoaraiosis. Dimethylarginine dimethylaminohydrolase (DDAH) is a hydrolytic enzyme that is primarily responsible for eliminating asymmetric dimethylarginine, and it plays a role in the pathogenesis of cardiovascular and cerebrovascular diseases. The DDAH2 subtype is expressed in organs rich in induced nitric oxide synthase, including the heart, the placenta, and the cerebral endothelium during cerebral ischemia, in the stress state, or under neurotoxicity. Overexpression of the DDAH2 gene can inhibit asymmetric dimethylarginine-induced peripheral circulating endothelial cell dysfunction. However, it is unknown whether this polymorphism regulates plasma asymmetric dimethylarginine levels in patients with leukoaraiosis. In this double-blind study, we recruited 46 patients with leukoaraiosis and 46 healthy, matched controls. Plasma asymmetric dimethylarginine levels were determined using enzyme-linked immunoassays. Genomic DNA was isolated from whole blood samples, and polymerase chain reaction, SmaI restriction enzyme digestion, restriction fragment length polymorphisms, and agarose electrophoresis were used to detect DDAH2 (-449 G/C) gene polymorphisms. The results revealed that 95.65% of leukoaraiosis patients had recessive genetic models (GG and CG), while 89.13% of healthy control subjects had dominant genetic models (CC and CG). There was a significant difference in the genotype composition ratio between leukoaraiosis patients and healthy controls (P = 0.0002). The frequency of G alleles in the leukoaraiosis patients (71.74%) was significantly higher than in healthy controls, whereas the frequency of C alleles was lower (χ2 = 13.9580, P = 0.0002). Furthermore, asymmetric dimethylarginine concentrations in subjects with the GG genotype were significantly higher than in subjects with the CG and CC genotypes (Kruskal-Wallis H = 24.5955, P < 0.0001). In addition, the GG genotype of DDAH2 (-449 G/C) was more common in patients with leukoaraiosis. These findings suggest that the G allele of DDAH2 (-449 G/C) is a risk factor for leukoaraiosis morbidity and is correlated with high levels of asymmetric dimethylarginine. This study was approved by the Institutional Ethics Committee of The 2nd Affiliated Hospital of Harbin Medical University of China (approval No. KY2016-177) on July 28, 2016.

LOC101930370/MiR-1471 Axis Modulates the Hedgehog Signaling Pathway in Breast Cancer.

BACKGROUND/AIMS: Non-coding RNAs (ncRNAs) play vital regulatory roles in many tumors. However, the functional roles of these transcripts responsible for their dysregulation in breast cancer (BC) are not thoroughly understood. METHODS: We examined the expression of microRNA miR-1471 in BC specimens. Online analysis tools predicted that lncRNA LOC101930370 might act as an endogenous 'sponge' by competing for miR-1471 binding targets. Luciferase assays were used to prove the interaction of LOC101930370, miR-1471 and SHH. Edu, wound-healing and transwell assays were used to verify the contribution of miR-1471 and LOC101930370 on MCF-7 cells proliferation and metastasis. Gain and loss of function studies were performed to evaluate the relevance of Hedgehog pathway with LOC101930370/miR-1471 regulating axis in MCF-7 cells. RESULTS: The expression of miR-1471 was markedly downregulated in BC. Inhibition of miR-1471 by LOC101930370 was proved by luciferase assay. Knockdown of LOC101930370 suppressed BC cells progression. MiR-1471 inhibitor resulted in a more aggressive metastasis of MCF-7 cells. Moreover, SHH and Gli-1 expression were significantly suppressed by LOC101930370 knockdown, and upregulated by miR-1471 inhibitor transfection. CONCLUSIONS: Collectively, our study reveals the interaction between LOC101930370 and miR-1471 for the first time. LOC101930370 positively regulates the expression of SHH by sponging miR-1471, which sheds new light on lncRNA-directed diagnostics and therapeutics in BC.

MiR-519d suppresses breast cancer tumorigenesis and metastasis via targeting MMP3.

Breast cancer (BC) is the most common cause of death in women throughout the world. Although microRNAs (miRNAs) have been identified as novel regulators in carcinogenesis, there are still abundant hidden treasure needed to be excavated. In the present study, we found that miR-519d expression was remarkably decreased in both human BC tissues and MCF-7 cells. CCK8 and 5-Ethynyl-2'-deoxyuridine (EdU) assays were used to evaluate cell proliferation. Wound-healing and transwell assays were performed for detection of cell migration and invasion. The results demonstrated miR-519d overexpression dramatically suppressed MCF-7 cells proliferation, migration and invasion. While downregulation of miR-519d by miR-519d inhibitor substantially increased MCF-7 cell carcinogenesis. Further analysis identified Matrix Metalloproteinase-3 (MMP3) as a direct target of miR-519d. QRT-PCR and western blot results indicated the correlative expression of miR-519d and MMP3 in BC tissues and MCF-7 cells. In summary, our data uncovered the novel molecular interaction between miR-519d and MMP3, indicating a therapeutic strategy of miR-519d for BC.

Analysis of risk factors in patients with leukoaraiosis.

To investigate the risk factors for leukoaraiosis (LA) and the correlation between risk factors and LA.The study comprised 92 patients with diagnoses of LA (LA group) and 56 non-LA individuals (control group). Data were collected for the following: age, gender, fasting blood glucose, total cholesterol, triglyceride, high- and low-density lipoprotein cholesterol, uric acid, creatinine, and histories of smoking, hypertension, and diabetes. Levels of serum asymmetric dimethylarginine (ADMA) were detected by enzyme-linked immunosorbent assay.Univariate analysis showed statistical significance between the 2 groups in age, histories of hypertension and smoking, uric acid, creatinine, and levels of serum ADMA (P < 0.05). Binary logistic analysis showed that age (P < 0.0001), hypertension (P = 0.0101), and serum ADMA (P = 0.0206) were related to LA. Pearson correlation analysis showed that levels of serum ADMA correlated with uric acid (r = 0.184, P = 0.025) and creatinine (r = 0.169, P = 0.04).Age, hypertension, and levels of serum ADMA were independent risk factors for LA. Serum ADMA levels may be related to uric acid and creatinine.

Protein kinase C delta mediated cytotoxicity of 6-Hydroxydopamine via sustained extracellular signal-regulated kinase 1/2 activation in PC12 cells.

OBJECTIVES: The incidence of Parkinson's disease (PD) is increasing as the global population ages. 6-hydroxydopamine (6-OHDA) can induce PD-like neuropathology and biochemical changes in both in vitro and in vivo models. Therefore, clarification of the molecular mechanism of 6-OHDA-induced cell death might contribute to the understanding of the pathogenesis of PD. METHODS: With this goal in mind, we investigated the role of protein kinase C delta (PKC delta) in 6-OHDA-dependent death using the pheochromocytoma cell line, PC12. Cells were treated with 6-OHDA to induce toxicity with or without pretreatment using rottlerin (a PKC delta inhibitor), bisindolylmaleimide I (a general PKC inhibitor), Gö6976 (a PKC inhibitor selective for calcium-dependent PKC isoforms), or phorbol-12-myristate-13-acetate (PMA, a PKC activator). RESULTS: Phorbol-12-myristate-13-acetate decreased cell survival and increased the rate of apoptosis while rottlerin increased cell survival and decreased the rate of apoptosis. In contrast, neither bisindolylmaleimide I nor Gö6976 affected 6-OHDA-induced cell death. Western analysis demonstrated that phosphorylation of PKC delta on Thr 505 as well as extracellular signal-regulated kinase (ERK) phosphorylation increased after exposure to 6-OHDA. This increase in PKC delta phosphorylation was potentiated by PMA. However, rottlerin attenuated the 6-OHDA-stimulated increase in PKC delta and ERK phosphorylation. CONCLUSION: These data suggest that PKC delta, rather than classic-type PKC (alpha, beta1, beta2, gamma), participates in 6-OHDA-induced neurotoxicity in PC12 cells, and PKC delta activity is required for subsequent ERK activation during cell death.

[AG490 inhibits the proliferation of K562 and down-regulates protein phosphatase PHLPP expression].

This study was aimed to investigate the effect of AG490, a JAK2 inhibitor, on expression of PHLPP and p-Akt in K562. K562 cells were treated with different concentrations of AG490. The proliferation of K562 cells was examined by WST-1 assay and apoptosis of K562 cells was detected by flow cytometry with Annexin V-FITC/PI double staining. The expressions of PHLPP, phosphorate-Akt (p-Akt) and total Akt protein were detected by Western blot. The results indicated that AG490 inhibited the proliferation of K562 cells in concentration-and time-dependent manners, with the IC(50) 338.0 µmol/L in 48 h. AG490 100 µmol/L also induced apoptosis of K562 cells in a time-dependent manner. AG490 100 µmol/L time-dependently down-regulated the protein expression of p-Akt and PHLPP, but without significant effect on expression of total Akt. It is concluded that AG490 can inhibit proliferation and induce apoptosis of K562 cells through down-regulation of p-Akt expression, but inhibiting efficacy of AG490 on K562 proliferation also may be limited due to the down-regulation of p-Akt regulatory protein PHLPP expression.

Depleting MEKK1 expression inhibits the ability of invasion and migration of human pancreatic cancer cells.

BACKGROUND: Mitogen-activated protein/ERK kinase 1 (MEKK1) is a Ser/Thr protein kinase belonging to the MEKK/STE11 subgroup of the MAPKKK family and plays a key role in tumor metastasis. However, it remains unclear about its functions in pancreatic cancer. MATERIALS AND METHODS: We analyzed MEKK1 expression in 41 surgically resection pancreatic cancer patient's samples by immunohistochemistry and determined its role in BxPC3 cells via RNAi experiment. The abilities of invasion, motility, and adhesion of BxPC3 cells were detected by transwell assay, wound healing assay and adhesion assay, respectively. Gelatinase activity of MMPs in cultured cells was examined by gelatin zymography. RESULTS: Our data showed that MEKK1 expression is positively correlated with lymphatic metastases (P < 0.01). The abilities of invasion, motility, and adhesion of BxPC3 cells were inhibited significantly (P < 0.01) when MEKK1 was depleted with a specific siRNA. We observed that the activity of MMP2 was decreased in the MEKK1 depletion cell line (P < 0.05), accompanied with decreased phosphorylated ERK1/2. CONCLUSION: Our results indicated that the depletion of MEKK1 led to a potent inhibition on the invasion and migration of human pancreatic adenocarcinoma in vitro. It suggests that MEKK1 may be a potential target for development of anti-invasion and metastasis drugs.

[Surgical treatment and prognosis of cancer of hepatic flexure of colon invading the duodenum in 65 patients].

OBJECTIVE: To discuss surgical treatment of right colon carcinoma of hepatic flexure invading the duodenum. METHODS: Sixty-five patients with right colon carcinoma of hepatic flexure invading the duodenum, treated in our department from 1987 to 2007, were included in this study. Their clinicopathological data were retrospectively reviewed and analyzed. All the cases were divided into three types (local invasion, regional invasion, and cancer with internal fistula) according to duodenal defect, including local invasion (< 2.0 cm), wide invasion (> 2.0 cm) and the presence of internal fistula. RESULTS: 25 patients with local invasion underwent en bloc resection of the duodenal wall. Pedicled ileal flap was used to cover the large duodenal defect measuring 2.0 - 3.0 cm in 5 patients. Dudenojejunostomy was used to reconstruct the large defect measuring more than 5 cm in 3 patients. Conservative resection of right-sided colon was performed in 18 patients with wide invasion. 4 patients underwent pancreaticoduodenectomy combined with right hemicolectomy for colon cancer involving the pancreatic head. 10 underwent duodenal diverticularization. One patient with anastomotic leakage healed within 3 weeks. Other patients were cured without postoperative complications. The total 3-year and 5-year survival rates after surgery were 53.8% and 9.2%, respectively. CONCLUSION: The surgical procedure to be performed is usually decided according to the cancer location, extent, and duodenal defect and invasion, which are important for prolonging life time, improving of quality of life and prognosis in these patients.

[Diagnosis and surgical treatment of carcinoid tumors of the appendix in 64 patients].

OBJECTIVE: To summarize the experience in the diagnosis and surgical treatment of carcinoid tumors of the appendix. METHODS: From 1972 to 2006, 64 patients with carcinoid tumors of the appendix received surgical treatment in our hospitals. The clinical data of those patients were retrospectively analyzed. RESULTS: Of the 64 cases, only 6 cases (9.4%) were correctly diagnosed preoperatively, while 58 (90.6%) not confirmed, with a misdiagnosis rate of 90.6%. All patients underwent surgical treatment, including appendectomy in 54, ileocecectomy in 4, right hemicolectomy in 2 and right hemicolectomy with regional lymph node dissection in 4 cases. The operation modes were determined according to the doctor's judgments based on the age of the patients, the nature, size, location, infiltration depth and lymph node metastasis of the tumors. Of the 64 patients, 58 were followed up with a longest follow-up period of 13 years, while 6 lost follow-up. Fifty-seven of those were still surviving, only one died of liver metastasis at 13 years after operation. CONCLUSION: Carcinoid tumor of the appendix is rare with a high rate of misdiagnosis before operation. Surgical resection is the only effective treatment for this disease and proper operation mode is the key to achieve good survival.

[Impact of trichostatin A on gastric carcinoma cell line SGC-7901].

OBJECTIVE: To investigate the effect of trichostatin A(TSA) on SGC- 7901 cells. METHODS: Cytotoxicity and cell viability of gastric cancer cell line SGC- 7901 were assayed by MTT method. Morphologic assessment of apoptosis was performed with fluorescence microscope. Cell cycle and apoptosis rate were analyzed by flow cytometry. Histone H3 acetylation was detected by Western blot. RESULTS: TSA showed apparently cytotoxicity in SGC- 7901 cells. The growth curve showed the growth ratio decreased with the increase of TSA concentration. Apoptosis rate were significantly different between TSA treated group(75 ng/ml for 72 h)and control group (P < 0.05). Morphologic changes of apoptosis including nuclear chromatin condensation and fluorescence strength were observed with fluorescence microscope.TSA treatment (75 ng/ml for 72 h) sensitively induced apoptosis in the cell,which was demonstrated by the migration of many cells to the sub- G1 phase,the reduction of G1- phase cells and the increment of apoptosis rate (29.54%) in flow cytometric analysis. The expression of acetylated histone H3 was increased in TSA group(75 ng/ml) for 48 h compared with control group by Western blot. CONCLUSIONS: TSA can induce SGC- 7901 cell apoptosis. The expression of acetylated histone H3 may contribute to the apoptosis.

[Clinical analysis of gastroparesis syndrome after nongastrectomy abdominal operation: a report of 22 cases].

OBJECTIVE: To investigate the causes, diagnosis and treatment of gastroparesis syndrome after nongastrectomy abdominal operation. METHODS: The clinical data of 22 cases with gastroparesis syndrome after nongastrectomy abdominal operation from 1972 to 2004 were retrospectively analyzed. RESULTS: Gastroparesis syndrome after nongastrectomy abdominal operation often occurred during 4-6 days postoperatively when the patients began to take in food, characterized by upper abdominal distension, nausea, vomiting, strong splashing bowel sound, weak bowel sound and large quantity of gastric drainage ranging from 1000 to 3000 ml every day. Barium meal was valuable not only in the diagnosis but also effective for promoting gas motility. It revealed a non-peristaltic, flabby and static stomach, and retention of contrast medium in the stomach even 5-6 hours later. All the patients recovered through non-operative therapy for 5-25 days including continuous gastrointestinal decompression, TPN and gastro-intestinal dynamic medicine. CONCLUSIONS: The causes of gastroparesis syndrome after nongastrectomy abdominal operation are multifactorial, most of such patients can be cured by non-operative therapy.

[Anatomic study of small intestinal vessel in piglet small intestine transplantation].

OBJECTIVE: To study the anatomy of the small intestine,and investigate the optimal selection of donors,recipients,and their small intestine vessels in piglet small intestine transplantation. METHODS: The weight and length of 30 piglets were measured. Angiography and pigments perfusion were used to observe the main vessels of the small intestine,and the length of the small intestine,and the external diameter of the main vessels of the small intestine were measured in vivo and ex vivo. RESULTS: The length of the small intestine was 11.5 times as long as the body length, and its weight accounted for 2.3% of the body weight. The outer diameters of abdominal aorta (AT), mesenteric anterior artery (MAA) and its 5(th)-6(th) branches in vivo and ex vitro were 4.3/4.6mm, 2.5/2.7mm and 1.9/2.2mm respectively. The total number of MAA's branches was 6-8 in general and its 5(th)-6(th) branches were the longest [(20.0 +/- 7.0) mm, (22.0 +/- 8.2) mm]. The outer diameter of mesenterial anterior vein (MAV) was 1-2 mm wider than that of MAA. CONCLUSIONS: AT, MAA and its 5(th)-6(th) branches are the preferable vessels for small intestine transplantation. In segmental small intestine transplantation, the length of the small intestine and body weight can be used to primarily select the suitable animals.

[Diagnosis and treatment of paraduodenal hernia: a report of 16 cases].

OBJECTIVE: To summarize the experience of diagnosis and treatment for paraduodenal hernia (PDH). METHODS: The clinical data of 16 cases with PDH surgically treated from 1962 to 2003 were analyzed retrospectively. RESULTS: The average age of the patients was 36 years old. There were 13 cases with left PDH (Landzerts hernia) and 3 cases with right PDH (Waldeyer's hernia). Ten patients usually had no symptoms, while acute abdominal pain occurred after full food in 3 case and after vigorous movement in 13 cases respectively. X-ray revealed complete or incomplete intestinal obstruction in 12 cases, while ultrasonography and CT revealed dilated cystic jejunal loop between the pancreas and the stomach in 6 cases. Fourteen cases (87.5%) were misdiagnosed before operation. The hospital stay ranged from 10 to 13 days. All patients were followed up from 2 to 4 years without recurrence. CONCLUSION: With high misdiagnostic rate, it is the key to perform exploratory operation for suspicious PDH.