High incidence of EDNRB gene mutation in seven southern Chinese familial cases with Hirschsprung's disease.

PURPOSE: Hirschsprung's disease (HSCR) is the leading cause of neonatal functional intestinal obstruction, which has been identified in many familial cases. HSCR, a multifactorial disorder of enteric nervous system (ENS) development, is associated with at least 24 genes and seven chromosomal loci, with RET and EDNRB as its major genes. We present a genetic investigation of familial HSCR to clarify the genotype-phenotype relationship. METHODS: We performed whole exome sequencing (WES) on Illumina HiSeq X Ten platform to investigate genetic backgrounds of core family members, and identified the possibly harmful mutation genes. Mutation carriers and pedigree relatives were validated by Sanger sequencing for evaluating the gene penetrance. RESULTS: Four familial cases showed potential disease-relative variants in EDNRB and RET gene, accounting for all detection rate of 57.1%. Three familial cases exhibited strong pathogenic variants as frameshift or missense mutations in EDNRB gene. A novel c.367delinsTT mutation of EDNRB was identified in one family member. The other two EDNRB mutations, c.553G>A in family 2 and c.877delinsTT in family 5, have been reported in previous literatures. The penetrance of EDNRB variants was 33-50% according mutation carries. In family 6, the RET c.1858T>C (C620R) point mutation has previously been reported to cause HSCR, with 28.5% penetrance. CONCLUSION: We identified a novel EDNRB (deleted C and inserted TT) mutation in this study using WES. Heterozygote variations in EDNRB gene were significantly enriched in three families and RET mutations were identified in one family. EDNRB variants showed an overall higher incidence and penetrance than RET in southern Chinese families cases.

Enhanced molecular binding affinity toward aromatic dications by anthracene-derived crown ethers in water.

The pursuit of high molecular binding affinity using conventional crown ethers in water remains a challenging task in the field of supramolecular chemistry and may hold great promise in the creation of advanced biocompatible nanoconstructs. In this work, the molecular binding strength toward a series of structurally relevant cationic guests has been greatly enhanced by tetrasulfonated 1,5-dianthracenyl-42-crown-10 and as investigated by means of 1H NMR, UV-vis, and fluorescence spectroscopy, the host-guest association constants can reach up to 108 M-1 order of magnitude in aqueous solution. X-ray crystal diffraction analysis further demonstrates that the aromatic dication can be tightly encapsulated in the ring of anthracene-derived crown ether via multiple π-stacking and electrostatic interactions. Meanwhile, the obtained association constants are remarkably higher than the ones in the cases of the known benzene- and naphthalene-derived sulfonated crown ethers, substantiating that the appropriate extension of π-conjugation in the molecular skeleton of crown ether is a feasible method in attaining a highly affiliative host-guest complex. Taken together, our results indicate that the anthracene-based sulfonated crown ether can be developed as a new family of water-soluble macrocyclic receptors in the fabrication of functional nanoarchitectures.

Effect of triptolide on expression of oxidative carbonyl protein in renal cortex of rats with diabetic nephropathy.

The traditional Chinese medicine (Tripterygium wilfordiiHook.f., TWH) has been clinically used to treat primary and secondary renal diseases and proteinuria for nearly 40 years. However, there is a rare literature about the effect of triptolide (the main active ingredient of TWH) on the expression of oxidative carbonyl protein (OCP) in diabetic nephropathy (DN). This study aimed to provide experimental evidence for triptolide treatment on DN through its effect on the expression of OCP, in order to investigate the effects of triptolide on the expression of OCP in rats with DN. Sixty SD rats were randomly divided into five groups: control group, high-dose triptolide (Th) group, low-dose triptolide (Tl) group, DN model group, and positive control (benazepril) group. The DN model was established using streptozotocin. Urinary protein excretion, fasting blood glucose (FBG), superoxide dismutase (SOD) in renal homogenate, malondialdehyde (MDA) in renal homogenate and renal nitrotyrosine by immunohistochemistry, and the expression of OCP by oxyblotimmune blotting were detected. In the DN model group, rat urinary protein excretion and renal MDA were significantly increased, while renal SOD significantly decreased and nitrotyrosine expression was obviously upregulated in the kidney. After triptolide treatment, 24-h urinary protein excretion (61.96±19.00 vs. 18.32±4.78 mg/day, P<0.001), renal MDA (8.09±0.79 vs. 5.45±0.68 nmol/L, P<0.001), and nitrotyrosine expression were decreased. Furthermore, renal OCP significantly decreased, while renal SOD (82.50±19.10 vs. 124.00±20.52 U/L, P<0.001) was elevated. This study revealed that triptolide can down-regulate the expression of OCP in the renal cortex of DN rats.

Effects and clinical significance of pentoxifylline on the oxidative stress of rats with diabetic nephropathy.

Diabetic nephropathy (DN) is a common and serious clinical complication of diabetes and presently there are no effective ways to prevent its occurrence and progression. Recent studies show that pentoxifylline (PTX) can improve renal hemodynamics, reduce urinary protein excretion, and alleviate or delay renal failure in DN patients. In this study, we focused on the anti-oxidative stress effect of PTX on alleviating renal damages of DN using rat models. DN rats were established with injection of streptozotocin. Blood glucose, urinary protein excretion, serum cystatin C, renal biopsy, superoxide dismutase (SOD) and malondialdehyde (MDA) in serum and renal homogenate and renal nitrotyrosine levels were analyzed before and 12 weeks after the treatment of PTX. Before treatment, all the DN rats had elevated blood glucose, increased urinary protein excretion and elevated serum cystatin C. Morphologically, DN rats exhibited renal tissue damages, including swelling and fusions of foot processes of podocytes under electron microscope. Masson staining revealed blue collagen deposition in glomeruli and renal interstitium. With treatment of PTX, symptoms and renal pathological changes of DN rats were alleviated. Furthermore, the MDA levels were increased and the SOD levels were decreased in the serum and kidneys of DN rats, and these changes were reversed by PTX. The expression of nitrotyrosine was up-regulated in DN rat model and down-regulated by PTX, indicating that PTX was able to inhibit oxidative reactions in DN rats. PTX could alleviate renal damage in DN, which may be attributable to its anti-oxidative stress activity.

Molecular characterization and gene functional analysis of Dicer-2 gene from Nilaparvata lugens (Hemiptera: Geometroidea).

Nilaparvata lugens (Stål) (Hemiptera: Geometroidea), a serious rice pest in many countries of Asia, causes a great loss in rice production every year. RNA interference (RNAi) is a powerful technology for gene function study in insects and a potential tool for pest control. As a core component of RNAi pathway, Dicer-2 (Dcr-2) protein determines the production of small interfering RNA (siRNA) and is crucial for the efficiency of RNAi. In this study, the full-length complementary DNA (cDNA) of N. lugens Dcr-2 (NlDcr-2) was first cloned and analyzed, and then the RNAi experiment was conducted to explore the function of NlDcr-2 gene. The complete Dcr-2 cDNA of N. lugens was 4 971 bp in length with an open reading frame (ORF) of 1,656 amino acids. Phylogenetic and protein domain analysis showed that the predicted NlDcr-2 protein was similar to Tribolium castaneum. In the RNAi experiment, the messenger RNA level of NlDcr-2 was significantly reduced by NlDcr-2 double-stranded RNA (dsRNA) (dsDcr-2). Fifty-five per cent decrease of NlDcr-2 was found after 4 days of unremitting feeding. No significant effect was observed on the development of N. lugens after dsRNA ingestion.

[Evaluation of scalene lymph node or contralateral mediastinum biopsy during mediastinoscopy for non-small cell lung cancer].

OBJECTIVE: To explore the clinical indication of N3 lymph node biopsy during mediastinoscopy for non-small cell lung cancer (NSCLC). METHODS: Cervical mediastinoscopy was performed in 89 patients with clinical stage I-IIIA non-small cell lung cancer prior to thoracotomy. Of those, 12 underwent cervical medistinoscopy combined with right scalene lymph node biopsy and 10 with anterior mediastinotomy. RESULTS: Nine patients were found to have lymph node metastasis (N3 disease) during mediastinosopy. Of those, 6 had contralateral mediastinal lymph node metastasis and 3 cases with right scalene lymph node metastasis. The incidence of N3 disease in the patients with adenocarcinoma, serum CEA > 5 ng/ml and multi-station mediastinal lymph node metastasis was significantly higher than that in those with non-adenocarcinoma, CEA < 5 ng/ml and ipsilateral uni-station mediastinal lymph nodes metastasis (P < 0.05). CONCLUSION: Biopsy of scalene lymph node or contralateral mediastinal lymph node should be performed during mediastinoscopy in order to exclude N3 disease for potentially operable NSCLC patients with adenocarcinoma, serum CEA >5 ng/ml and ipsilateral multi-station mediastinal lymph nodes metastasis.

[Value of mediastinoscopy in preoperative staging of non-small cell lung cancer-based on survival analysis].

OBJECTIVE: To evaluate the value of mediastinoscopy in preoperative staging of non-small cell lung cancer (NSCLC) based on survival analysis. METHODS: 152 cases of potentially operable NSCLC were enrolled in this study. All cases underwent CT scan and mediastinoscopy for assessment of the mediastinal lymph node status before initial treatment. The definitive treatment was decided on the basis of mediastinoscopy and the survival rate was analyzed with a median follow-up of 30.5 months. Survival analysis was conducted by comparing the lymph node status which was determined by final pathology (groups pN0, pN1, pN2, pN3), CT scan (group cN0-1, cN2-3) and mediastinoscopy (group mN0-1, mN2, mN3). RESULTS: The 5-year survival rates in group pN0, pN1, pN2 and pN3 were 61.7%, 75.0%, 32.4% and 16.1%, respectively. Both groups pN0 and pN1 had significantly higher survival rates than those in groups pN2 and pN3 (P < 0.05). There were not significant differences between survival rates in groups cN0-1 and cN2-3 (P = 0.670), while the survival rate in group mN0-1 was significantly higher than that in groups mN2 and mN3 (P < 0.05). CONCLUSION: Mediastinoscopy is of great value in preoperative staging of NSCLC. Not only does it detect lymph node metastasis more precisely but also better predict the prognosis than CT scan.