[Differential Expression of Long Non-coding RNA Cancer Susceptibility Candidate 2 and Imprinted Gene H19 in Extrahepatic Cholangiocarcinoma].

Objective To investigate the expression and the potential roles of long non-coding RNA(lncRNA)cancer susceptibility candidate 2(CASC2)and imprinted gene H19 in extrahepatic cholangiocarcinoma(ECC). Methods Four samples from patients with ECC were collected for high-throughput sequencing which was conducted to reveal the transcriptomic profiles of lncRNA CASC2 and H19.Bioinformatics tools were employed to predict the potential roles of the two genes.Another 22 ECC tissue samples and the cholangiocarcinoma cell lines(RBE,QBC939,HuH-28,and HuCCT1)with different degrees of differentiation were selected for validation.The para-carcinoma tissue and normal human intrahepatic biliary epithelial cell(HIBEC)were used as the control groups.The expression levels of lncRNA CASC2 and H19 in carcinoma tissue,para-carcinoma tissue,and cell lines were determined by real-time quantitative polymerase chain reaction(qRT-PCR).The correlation analysis was carried out for the clinical indicators of patients with the expression levels of the target genes. Results The two target genes showed significantly different expression between carcinoma tissue and para-carcinoma tissue(all P<0.05).Specifically,CASC2 had higher expression level in the carcinoma tissue than in the para-carcinoma tissue(t=1.262,P=0.025),whereas the expression of H19 showed an opposite trend(t=1.285,P=0.005).The expression levels of CASC2 in QBC939(t=8.114,P=0.015)and HuH-28(t=9.202,P=0.012)cells were significantly higher than that in the control group.The expression levels of H19 were significantly lower in RBE(t=-10.244,P<0.001),QBC939(t=-10.476,P<0.001),HuH-28(t=-19.798,P<0.001),and HuCCT1(t=-16.193,P=0.004)cells than in the control group.Bioinformatics analysis showed that CASC2 was mainly involved in the metabolic process and H19 in the development of multicellular organisms.Both CASC2 and H19 were related to catalytic activity.The expression level of lncRNA CASC2 was correlated with pathological differentiation(χ 2=6.222,P=0.022)and lymph node metastasis(χ2=5.455,P=0.020),and that of lncRNA H19 with pathological differentiation(χ2=1.174,P=0.029)and tumor size(χ2=-0.507,P=0.037). Conclusions In the case of ECC,lncRNA CASC2 and H19 have transcription disorders.lncRNA CASC2 is generally up-regulated in the carcinoma tissue,while H19 is down-regulated.Both genes have the potential to become new molecular markers for ECC.

Pathway of Diatoms Enter Experimental Rabbits through the Lymphatic System of the Digestive Tract.

OBJECTIVES: To study whether diatoms can enter the body through the lymphatic system of the digestive tract. METHODS: Twenty experimental rabbits were divided into the test group and the control group randomly, and intragastric administration was performed with 20 mL water sample from the Pearl River and 20 mL ultrapure water, respectively. After 30 min, lymph, lungs, livers and kidneys were extracted for the diatom test. The concentration, size and type of diatoms were recorded. RESULTS: The concentration of diatoms of the test group was higher than that of the control group (P<0.05). In the test group, Stephanodiscus, Coscinodiscus, Cyclotella, Melosira, Nitzschia, Synedra, Cymbella, and Navicula were detected; in the control group, Stephanodiscus, Coscinodiscus and Cyclotella were detected. The long diameter and the short diameter of diatoms of the test group were higher than those of the control group (P<0.05). In the test group, 1-2 diatoms were detected in 3 lung samples and 2 liver samples, which were Stephanodiscus or Cyclotella, and no diatoms were detected in the kidney samples; in the control group, 1-2 diatoms were detected in 2 lung samples and 3 liver samples, which were Stephanodiscus or Coscinodiscus, and no diatoms were detected in the kidney samples. CONCLUSIONS: Diatoms can enter the body through the lymphatic fluid, which is one of the reasons for the presence of diatoms in tissues and organs of non-drowning cadavers.

[Male infertility - related gene knockout mouse models].

The pathogenesis of male infertility is rather complicated. The establishment of animal models, especially mouse models, of male infertility, provides a model basis for the studies of the roles and molecular mechanisms of infertility-related genes. Currently there are mainly three types of mouse models for biomedical researches, namely, the mouse model made by the knockout, knock-in or gene capture method, transgenic mouse model, and chemically induced point mutant mouse model. This review summarizes male infertility - related gene knockout mouse models, aiming to find a suitable animal model for studying the pathogenesis of male infertility.

Identification and characterization of a novel Bacillus subtilis strain with potent antifungal activity of a flagellin-like protein.

The filamentous fungus Botrytis cinerea is an important agricultural pathogen affecting a wide range of cultivated plants. Since World War II, chemical fungicides have been the go-to method for agricultural pathogen control. However, the potential adverse environmental and health effects of these chemicals have led to an increasing demand for alternative methods of pathogen control, including biological control agents. In this study, we identified a bacterial isolate with strong antagonistic activity against B. cinerea. An analysis of the 16S rRNA gene sequence for this isolate identified it as a novel strain of Bacillus subtilis. Culture media from this isolate were harvested and fractionated using ion exchange and gel filtration chromatography. The fraction exhibiting the highest level of antifungal activity was identified, and its sequence determined by electrospray tandem mass spectrometry had significant similarity to flagellin. This flagellin-like protein was exogenously expressed in Escherichia coli, and screened for antifungal activity against B. cinerea. This flagellin-like protein demonstrated clear antifungal activity of inhibiting B. cinerea growth.

The Controversy of Pepsinogen A/Pepsin A in Detecting Extra-Gastroesophageal Reflux.

BACKGROUND: Pepsinogen A (PGA)/pepsin A is often used as a diagnostic marker of extra-gastroesophageal reflux. We aimed to explore whether its positivity in upper aerodigestive tract (UADT) was specific enough to diagnose reflux. METHODS: PGA/pepsin A protein levels were examined in 10 types of tissues and 10 types of body fluid by immunological staining, western blot or Elisa, using three different commercially available brands simultaneously. Liquid chromatography-tandem mass spectrometry parallel reaction monitoring (LC-MS/MS PRM) served as a gold reference for the detection of PGA/pepsin A proteins. PGA gene expression was analyzed by reverse transcriptase sequencing methods for tissue samples. Specifically, 24 hour pH monitoring technique was conducted for patients who donated saliva samples. RESULTS: Eight out of ten types of human tissue samples (stomach, esophagus, lung, kidney, colon, parotid gland, nasal turbinate and nasal polyps) were confirmed positive for PGA/pepsin A gene and protein by genetic and PRM technique, respectively. Two out of ten types of body fluid samples (gastric fluid, urine) were confirmed positive for PGA/pepsin A protein by PRM technique. The consistence rates of PGA/pepsin A positivity among three commercial antibody brands and Elisa kit were poor, and Elisa results of salivary did not match with 24-hour pH monitoring. CONCLUSIONS: Multiple tissues and body fluid could be detected baseline expression levels of PGA/pepsin A gene and protein. However, those commercially available PGA/pepsin A antibodies achieved poor sensitivity and specificity, therefore, relying on the detection of PGA/pepsin A in UADT by single antibodies to diagnose extra-gastroesophageal reflux without a specific positive cut-off value is unreliable.

Establishment of a Canine Training Model for Digestive Tract Reconstruction after Pancreaticoduodenectomy.

PURPOSE: Practical training models can be a viable and effective educational tool that allows surgeons to acquire specific surgical techniques or skills. However, a suitable animal training model for reconstruction after a pancreaticoduodenectomy (PD) has not yet been reported. Therefore, we explored the feasibility and safety of establishing an animal training model for digestive tract reconstruction after a simulated PD using mongrel dogs. METHODS: We used the anatomical similarity between the canine and human digestive tract to simulate the digestive tract reconstruction after pancreatoduodenectomy. A hepatobiliary surgeon performed simulated PD digestive reconstructions on 6 mongrel canines. Pancreaticojejunostomy (PJ), biliary-enteric anastomosis (BEA), and jejuno-jejunal anastomosis (JJ) were performed sequentially. The survival rate, surgical operation time, complications, body weight changes, gross specimen, and pathological examination of the anastomotic region were observed 30 days after surgery. RESULTS: The survival rate 30 days after surgery was 100%. Total mean operative time was 230.5 ± 39.7 min. The operative time for PJ, BEA, and JJ was calculated as 21.5 ± 7 min, 21.7 ± 8.7 min, and 13.2 ± 1.8 min, respectively. An incision infection occurred in 1 case (16.7%); there was 1 case of ascites (16.7%), and 1 case of vomiting (16.7%). The total protein and total bilirubin indicators of the 6 dogs and the serum amylase index of 5 dogs 30 days postoperatively were within the normal range. The 6th dog's serum amylase was approximately double the normal value, possibly due to pancreatitis. Observing the gross specimen, the mucosa of the anastomosis was intact and smooth. Masson staining showed that the bile duct and jejunum anastomosis, the pancreas, and jejunum of the 6 canines were all integrated with rich collagen. CONCLUSION: Establishing an animal model for digestive tract reconstruction after a simulated PD in canines is feasible and safe.

Endoscopic linear stapler-assisted resection of a giant solid pseudopapillary pancreatic tumor with concurrent regional portal hypertension: a case report.

Solid pseudopapillary tumor of the pancreas (SPTP) is a rare neoplasm with a low incidence and low rate of malignancy. We herein report a rare case of SPTP concurrent with regional portal hypertension (RPH) that was successfully treated by distal pancreatectomy and splenectomy. A 22-year-old woman presented with a left upper abdominal apophysis and normal liver function. She was diagnosed with an SPTP and RPH by abdominal ultrasound and computed tomography, and she subsequently underwent distal pancreatectomy and splenectomy. Noticeably, varicose vein plexus with wide range appeared on the upper edge of the pancreatic body and posterior gastric wall of the patient. Therefore, we created a path to avoid touching the varicose veins and took advantage of the endoscopic linear stapler to staple the veins. We herein report our surgical experience on SPTP assisted with the endoscopic linear stapler, which will be very realistic for the management of this rare clinical entity.

Mirizzi syndrome complicated by common hepatic duct fistula and left hepatic atrophy: a case report.

BACKGROUND: Mirizzi syndrome is a rare complication of chronic cholecystitis, usually caused by gallstones impacted in the cystic duct or the neck of the gallbladder. Mirizzi syndrome results in compression of the hepatic duct or fistula formation between the gallbladder and common bile duct (or hepatic duct, right hepatic duct, or even mutative right posterior hepatic duct). Clinical features include abdominal pain, fever, and obstructive jaundice. Severe inflammation and adhesion at Calot's triangle are potentially very dangerous for patients with Mirizzi syndrome undergoing cholecystectomy. Case presentation: We report the case of a 68-year-old Asian woman who presented with abdominal pain and jaundice. She had a medical history of gallstones, but no fever. Magnetic resonance cholangiopancreatography revealed cholecystitis, cholelithiasis, common hepatic duct stones, and ascites. Findings at surgery included a porcelainized, atrophic gallbladder that was full of gallstones, fistula formation between the gallbladder and common hepatic duct, and left hepatic atrophy. The prominent feature was the left hepatic atrophy, but stones were not visible pre-operatively in the left liver by radiologic examination. CONCLUSIONS: This patient exhibited what can be considered a special type II of Mirizzi syndrome with a fistula of the common hepatic duct as well as left hepatic atrophy.

Modified laparoscopic cholecystectomy technique for treatment of situs inversus totalis: A case report.

Laparoscopic cholecystectomy is a broadly used technique for gallbladder treatment. However, situs inversus, a rare anomaly, is reportedly difficult to treat by conventional laparoscopic cholecystectomy. A 36-year-old woman with chronic cholecystitis and multiple gallstones was found to have dextrocardia on a chest X-ray. Magnetic resonance imaging demonstrated situs inversus, cholecystitis, and cholelithiasis. We successfully performed laparoscopic cholecystectomy using our modified technique, which mainly involved a left-handed operation and adjustment of the port positions. This case will be very instructive for right-handed surgeons in the management of cholelithiasis by laparoscopic cholecystectomy in patients with situs inversus.

PepsinA as a Marker of Laryngopharyngeal Reflux Detected in Chronic Rhinosinusitis Patients.

Objectives We aimed to confirm the presence of pepsinA in the nasal secretions and tissues of chronic rhinosinusitis (CRS) patients and reveal the relationship between CRS and laryngopharyngeal reflux (LPR). Study Design Cross-sectional study. Setting The study was conducted at the Department of Oto-Rhino-Laryngology, West China Hospital, Sichuan University. Subjects and Methods A total of 32 CRS patients with or without nasal polyps (CRSwNP and CRSsNP, respectively) and 10 normal controls were enrolled in our study. We investigated the expression of pepsinA in the nasal tissues, secretions, and blood plasma from the subjects by immunohistochemical staining, Western blot, or ELISA. Additionally, the expressions of MUC4, MUC5AC, MUC5B, MUC8, and pepsinogenA in nasal tissue were evaluated by quantitative real-time polymerase chain reaction. Results Immunohistochemistry and Western blot revealed that the pepsinA expression levels in the turbinate mucosa in CRSwNP/CRSsNP patients, which were largely restricted to the epithelial layer or glandular mucous cells in nasal tissues, were significantly higher than those in controls and in the polyp tissues of CRSwNP patients ( P < .05). In addition, the concentration of pepsinA in nasal secretions was significantly increased in the CRSwNP (147.85 ± 53.69 ng/mL, P < .001) and CRSsNP (134.12 ± 36.23 ng/mL, P < .001) groups as compared with the controls (68.69 ± 19.28 ng/mL). Although MUC5AC, MUC5B, and MUC8 expression differed among the groups, no correlation between pepsinA and mucin genes was found. Conclusion The results of this study provided evidence of an association between LPR and CRS, although no correlation was found to exist between LPR and mucin genes in CRS patients.

The Wnt/ß-catenin signaling pathway mechanism for pancreatic cancer chemoresistance in a three-dimensional cancer microenvironment.

ß-catenin is a key protein that is encoded by the CTNNB1 gene in the Wnt signaling pathway. This study investigated the associations between ß-catenin expression and implications for the efficacy of gemcitabine on pancreatic cancer cells in a three-dimensional (3-D) cancer microenvironment. For low ß-catenin expression pancreatic carcinoma cells, the inhibition rates (IRs) for low, middle, and high doses of gemcitabine were 0.615 ± 0.079, 0.691 ± 0.093, and 0.765 ± 0.061, respectively. For the high ß-catenin expression pancreatic carcinoma cells, the IRs for the same doses were 0.325 ± 0.072, 0.453 ± 0.075, and 0.537 ± 0.056, respectively. Additionally, the evaluation of ß-catenin immunoreactivity in 31 pancreatic cancer patients revealed that the low ß-catenin protein expression group had significantly longer overall survival (OS) and disease free survival (DFS) than the high ß-catenin protein group (P < 0.05). Overall, ß-catenin protein expression levels were significantly correlated to gemcitabine sensitivity in seven pancreatic carcinoma cell lines in the 3-D cancer microenvironment. These data suggest that large-scale clinical studies are warranted to assess the role of the Wnt/ß-catenin signaling pathway on ß-catenin protein expression and chemosensitivity to gemcitabine in pancreatic cancer.

Studied microRNA gene expression in human hepatocellular carcinoma by microRNA microarray techniques.

AIM: To achieve a better understanding of the molecular mechanisms of microRNA expression changes involved in hepatocellular carcinoma. METHODS: In this research process, patients were not treated with antivirals, immunosuppressants or immunomodulators for at least 6 mo before collecting serum. The study population was composed of 35 outpatient hepatitis B virus (HBV) cases and 12 healthy control cases from the Affiliated Hospital of Inner Mongolia Medical University (Inner Mongolia, China) from July 2013 to April 2014. The 35 HBV cases were divided into two groups: a hepatocirrhosis group with 20 cases and a liver cancer group with 15 cases. All 35 cases carried HBsAg. The diagnostic criteria followed the European Association for the Study of the Liver 2012 (EASL2012) standards. MicroRNA (miRNA) was extracted from a control group of patients, a group with hepatocirrhosis and a group with liver cancer and its quality was analyzed using the human V2 microRNA expression beadchip. Cluster analysis and a radar chart were then applied to the miRNA changes. RESULTS: The miRNA-qualified rate of human serum samples was 93%. The concentration of a single sample was > 200 ng/µL and the volume was > 5 µL. All miRNA serum samples were uncontaminated by the genome. The Mann-Whitney test showed significant differences in miRNA between each group, with a detection P-value of < 0.05. Illumina software was set up with Diff Score set to ± 13, meaning that P = 0.001.There were significant changes in miRNA expression between the three groups. miRNA-183 was the most up-regulated, followed by miRNA-373. miRNA-129 and miRNA-188 were both strongly down-regulated and miRNA-378 was down-regulated a small amount. The liver cancer group had greater changes, which indicated that changes in miRNA expression levels were caused by hepatocirrhosis. The liver cancer disease course then further increased these changes. In the pentagon created by these five miRNAs, three groups showed significant deviation. The liver cancer group had a bigger deviation trend. The chart indicated that miRNA expression changes occurred in the hepatocirrhosis group, which increased in the liver cancer disease course and were irreversible. CONCLUSION: There was a significant relationship between the irreversible up-regulation of miRNA-183/373 and down-regulation of miRNA-129/188/378 and incidences of hepatocirrhosis and liver cancer.

Langerhans cell sarcoma arising from the root of tongue: a rare case.

Langerhans cell sarcoma (LCS), a rare malignant disease with markedly malignant cytological features and poor outcome, originates from Langerhans cells and most commonly affects the lymph nodes, skin, and bone. This paper presents the case of a 58-year-old female with LCS at the root of her tongue, with neither local recurrence nor distant metastasis observed during 47 months of follow up following radiotherapy for more than one month after complete tumor resection. Histological and immunophenotypic tests revealed that the malignant tumor cells were positive for S-100 protein, CD1a, and LCA, and partially positive for CD3ε. By contrast, the tumor cells were negative for langenin, CD30, HMB45, PCK, CK5/6, and P63. Their Ki-67proliferation index ranged from 30% to 40%. This neoplasm was diagnosed as LCS according to the classification of WHO2008. This work is the first report on LCS arising from the root of tongue. This rare case may serve as a reference for future clinical studies.

Effects of Bifidobacterium Breve Feeding Strategy and Delivery Modes on Experimental Allergic Rhinitis Mice.

BACKGROUND: Different delivery modes may affect the susceptibility to allergic diseases. It is still unknown whether early intervention with probiotics would counteract this effect. OBJECTIVES: The effect of different delivery modes on immune status and nasal symptoms was investigated on established allergic rhinitis (AR) mouse model. In addition, the immunoregulatory effects and mechanisms of different feeding manners with Bifidobacterium breve(B. breve) were examined. METHODS: Live lyophilized B. breve was orally administered to BALB/c mice born via vaginal delivery(VD) or cesarean delivery (CD) for 8 consecutive weeks, after which they were sensitized by ovalbumin(OVA) to establish experimental AR. Nasal symptoms, serum immunoglobulins, cytokines, splenic percentages of CD4(+)CD25(+)Foxp3(+) regulatory T(Treg) cells and nasal eosinophil infiltration were evaluated. RESULTS: Compared with VD mice, mice delivered via CD demonstrated more serious nasal symptoms, higher concentrations of OVA-specific immunoglobulin (Ig) E, more nasal eosinophils and lower percentages of splenic CD4(+)CD25(+)Foxp3(+)Treg cells after establishing experimental AR. These parameters were reversed by administering B. breves hortly after birth. However, the effect of B. breve did not differ between different delivery modes. CONCLUSION: CD aggravates the nasal symptoms of AR mice compared to VD. This is the first report that oral administration of B. breve shortly after birth can significantly alleviate the symptoms of AR mice born via both deliveries, probably via activation of the regulatory capacity of CD4(+)CD25(+)Foxp3(+)Treg cells.