Transcription factor PbNAC71 regulates xylem and vessel development to control plant height.

Dwarfism is an important agronomic trait in fruit breeding programs. However, the germplasm resources required to generate dwarf pear (Pyrus spp.) varieties are limited. Moreover, the mechanisms underlying dwarfism remain unclear. In this study, "Yunnan" quince (Cydonia oblonga Mill.) had a dwarfing effect on "Zaosu" pear. Additionally, the dwarfism-related NAC transcription factor gene PbNAC71 was isolated from pear trees comprising "Zaosu" (scion) grafted onto "Yunnan" quince (rootstock). Transgenic Nicotiana benthamiana and pear OHF-333 (Pyrus communis) plants overexpressing PbNAC71 exhibited dwarfism, with a substantially smaller xylem and vessel area relative to the wild-type controls. Yeast one-hybrid, dual-luciferase, chromatin immunoprecipitation-qPCR, and electrophoretic mobility shift assays indicated that PbNAC71 downregulates PbWalls are thin 1 expression by binding to NAC-binding elements in its promoter. Yeast two-hybrid assays showed that PbNAC71 interacts with the E3 ubiquitin ligase PbRING finger protein 217 (PbRNF217). Furthermore, PbRNF217 promotes the ubiquitin-mediated degradation of PbNAC71 by the 26S proteasome, thereby regulating plant height as well as xylem and vessel development. Our findings reveal a mechanism underlying pear dwarfism and expand our understanding of the molecular basis of dwarfism in woody plants.

[Chemical constituents of cyclic peptides from fibrous roots of Pseudostellaria heterophylla].

Four cyclic peptides were isolated from the 75% ethanol extract of the fibrous roots of Pseudostellaria heterophylla by silica gel, Sephadex LH-20 column chromatography, and semi-preparative HPLC. Through mass spectrometry, NMR and other methods, they were identified as pseudostellarin L(1), heterophyllin B(2), pseudostellarin B(3), and pseudostellarin C(4). Among them, compound 1 was a new cyclic peptide, and compounds 2-4 were isolated from the fibrous roots of P. heterophylla for the first time. None of these compounds displayed cytotoxic activities against MCF-7, A549, HCT-116, and SGC-7901 cells.

A new cyclic peptide from the fibrous root of Pseudostellaria heterophylla.

A new cyclic peptide, Pseudostellarin K (1), together with thirteen known compounds, including two cyclic peptides (2 and 3), one ß-carboline alkaloid (4), two amides (5 and 6), three phenylpropanoids (7-9) and other compounds (10-14), were isolated from the fibrous root of Pseudostellaria heterophylla. Their structures were elucidated by extensive spectroscopic analysis. Compounds 1, 4-6, 10 were isolated from the genus pseudostellaria for the first time. All compounds were evaluated for cytotoxic activities against MCF-7, A549, HCT-116 and SGC-7901 cell lines by MTT assay. Unfortunately, all these compounds displayed weak cytotoxic activities.

Circulating MicroRNA-122 for the Diagnosis of Hepatocellular Carcinoma: A Meta-Analysis.

BACKGROUND: Circulating microRNA-122 (miR-122) has been recognized as a marker of hepatocellular carcinoma (HCC). The current meta-analysis was performed to quantitatively evaluate the diagnostic performance of circulating miR-122 for HCC. METHODS: Related studies that evaluated the diagnostic performance of circulating miR-122 determined from pathophysiological examination for HCC were obtained by systematic searches of the PubMed and Embase databases. A randomized fixed effects model was applied according to the heterogeneity among studies. The pooled sensitivity, specificity, and area under the summary receiver operating characteristic curve (AUC) were calculated to evaluate the diagnostic accuracy. Publication bias was detected by Deeks' funnel plot asymmetry test. RESULTS: Thirteen studies providing data for 920 HCC patients and 1217 controls were included in the meta-analysis. The pooled sensitivities, specificities, and AUCs of serum miR-122 were 0.76, 0.75, and 0.82, respectively, for discriminating HCC patients from overall controls; 0.85, 0.83, and 0.91, respectively, for discriminating HCC patients from healthy controls; 0.79, 0.82, and 0.87, respectively, for discriminating HCC from HBV or HCV infection; and 0.65, 0.75, and 0.74, respectively, for discriminating HCC from liver cirrhosis or dysplastic nodule formation. No significant publication bias was detected. CONCLUSIONS: Serum miR-122 confers moderate efficacy for discriminating HCC patients from healthy controls or patients with HBV or HCV infection, but not for discriminating HCC patients from those with liver cirrhosis or dysplastic nodule formation.

MicroRNA-497 functions as an inflammatory suppressor via targeting DDX3Y and modulating toll-like receptor 4/NF-κB in cigarette smoke extract-stimulated human bronchial epithelial cells.

BACKGROUND: We aimed to investigate the biological effect of miR-497 in cigarette smoke extract (CSE)-damaged human bronchial epithelial (HBE) cells and the underlying molecular mechanism. METHODS: MiR-497 mimic was transfected into HBE cells to up-regulate miR-497 expression. Cigarette smoke extract (CSE, 20 µg/mL) was utilized to treat HBE cells to form the injury model. Cell proliferation and apoptosis were detected by CCK8 and flow cytometry assays. DDX3Y mRNA expression was determined by a quantitative reverse transcriptase-polymerase chain reaction. The interaction between miR-497 and DDX3Y was verified by a luciferase reporter assay. Protein expression levels were tested by western blotting. RESULTS: CSE treatment decreased miR-497 level in HBE cells. CSE exposure restrained cell proliferation, promoted cell apoptosis and enhanced the relative expression of TLR4 and p-NF-κB p65. DDX3Y was predicted as a target of miR-497. The mRNA and protein expression of DDX3Y was negatively modulated by miR-497 in CSE-injured HBE cells. Up-regulation of miR-497 by miR-497 mimic increased cell proliferation and reduced cell apoptosis in CSE-treated HBE cells, which were rescued by DDX3Y high expression in CSE-treated HBE cells. Consistently, Bcl-2 protein level was heightened, whereas Bax and actived caspase-3/9 protein levels were decreased by miR-497 mimic in CSE-stimulated HBE cells, which was reversed by DDX3Y over-expression in CSE-stimulated HBE cells. The relative expression of TLR4 and p-NF-κB p65 was decreased by miR-497 mimic, whereas they were rescued by DDX3Y over-expression in CSE-damaged HBE cells. CONCLUSIONS: The results of the present study demonstrate that up-regulation of miR-497 exhibits a protective effect on CSE-damaged HBE cells, which might be achieved by targeting DDX3Y and regulating the TLR4/NF-κB pathway.

Gastrodin protects retinal ganglion cells through inhibiting microglial-mediated neuroinflammation in an acute ocular hypertension model.

AIM: To investigate the neuroprotective effect of gastrodin on retinal ganglion cells (RGCs) in an acute ocular hypertension (AOH) rat model and to identify its possible mechanism. METHODS: AOH rat model was performed in a randomly selected eye by anterior chamber perfusion and either received an intraperitoneal injection with various concentrations of gastrodin or normal saline. After 2wk, the rats were sacrificed. FluoroGold was used to label survival RGCs. Immunostaining with anti-Iba1 in the retinal flat mounts to calculate the microglia density in the ganglion cell layer (GCL). Changes in microglial cytokines, tumour necrosis factor-alpha (TNF-α) and inducible NO synthase (iNOS) were examined with Western blot and reverse transcription-quantitative polymerase chain reaction. Expression levels of total and phosphorylated p38 mitogen activated protein kinase (MAPK) were determined by Western blot. RESULTS: Results showed that AOH induced significant loss of RGCs and severe microglia activation in the GCL. Besides, AOH increased the phosphorylation of p38 MAPK and promoted the release of microglial cytokines in the retinas. Intraperitoneal injection with dose-dependent gastrodin significantly reduced the loss of RGCs and inhibited retinal microglia activation, accompanied with the decreased expression levels of microglial cytokines and p38 MAPK phosphorylation. CONCLUSION: Gastrodin exerts a neuroprotective effect on RGCs in an acute glaucoma animal model via inhibiting microglia activation and microglial-mediated neuroinflammation. The finding demonstrates the potential application of gastrodin in the neuroprotective therapy of acute glaucoma and other retinal neurodegenerative diseases characterized by microglia activation and RGCs death.

[Thumb replantation through bridging ulnar proper digital artery and dorsal carpal branch of radial artery].

OBJECTIVE: To introduce treatment method of replantation of severed thumb trauma with proximal arterial, and to evaluate its efficacy. METHODS: From February 2007 to March 2009,13 patients with severed thumb in serious injury of proximal arterial were treated with vein graft of volar forearm,bridging between dorsal carpal branch of radial artery on nasopharyngeal fossa and distal stump of ulnar proper digital artery. Among them, there were 11 males and 2 females with an average age of 34.5 years ranging from 16 to 50 years. Seven cases were in the left thumb,6 in the right thumb. Eight cases were complete separation, incomplete separation in 5 cases. Ten cases recieved emergency reimplantation, 3 cases with the arterial crisis after conventional replantation were explorated and repaired. RESULTS: Thumb of 13 cases all survived. All patients were followed-up for 8 to 17 months (averaged, 11 months). Replantation thumb obtained satisfactory appearance. According to Chinese Medical Association Society of Hand Surgery Trial criteria, the results were excellent in 9 fingers, good in 3 fingers, 1 poor finger. Two point discrimination of finger pulp was 5 to 8 mm (averaged 6.5 mm). CONCLUSION: By repairing artery of thumb with vein graft of volar forearm, bridging between dorsal carpal branch of radial artery on nasopharyngeal fossa and distal stump of ulnar proper digital artery,complex severed thumb replants on surgery position comfortable,without affecting the blood supply of the hand, expanding the indications for replantation and improving the success rate of replantation.

[Treatment of carpal tunnel syndrome with mini-incision decompression].

OBJECTIVE: To introduce the method of the transverse carpal ligament (TCL) transection through palmar mini-incision for carpal tunnel syndrome (CTS) and evaluate its efficacy and safety. METHODS: From January 2006 to September 2007, 15 patients (18 hands) with CTS were treated with palmar longitudinal mini-incision decompression. There were 2 males and 13 females, with an average age of 48 years (ranged 34 to 69) and an average course of 18 months (ranged 8 to 26). The main clinical symptom including the radial numbness or pain of the 3 and a half fingers, wrist pain and radiation to the forearm, the night awake history of numbness, thenar muscle atrophy, positive Tinel sign and Phalen sign. Median nerve electrophysiology showed that sensory nerve conduction velocity (SCV) slowed down and sensory nerve action potential (SNAP) decreased or missed, short abductor muscle of thumb had spontaneous potential in severe cases. Clinical effect were evaluated according to the Global symptom score (GSS) scoring in aspect of pain, numbness, paraesthesia, weakness and nocturnal awakening. RESULTS: Postoperative wound smoothly healed in all patients and no complications occurred. All patients were followed up from 20 to 28 months with an average of 24 months. Symptoms of 1 patient had not incomplete relief, other patient's symptoms disappeared and muscle force of abductor pollicis brevis reinforced. Postoperative GSS scoring obviously improved than preoperative (P < 0.05). CONCLUSION: Treatment of carpal tunnel syndrome through palmar mini-incision decompression has advantages such as higher safe, shorter operative time, less invasive, smaller scar, which can cut off carpi transversum ligament and thoroughly decompress median nerve under direct sight, it is a safe and effective operative approach.

[Consideration on relative issues of gastrointestinal stromal tumor].

Nowadays one of the hot research topics in gastrointestinal surgery is gastrointestinal stromal tumor (GIST). However, mechanisms of their formation and development, and the causes of recurrence following R0 resection, have not been fully understood. This article aims to investigate some important issues concerning the neoplastic essences of GIST including the genomics, bionomics, drug resistance and immunotherapy.

[Analysis of gene expression patterns in an irinotecan-resistance colon cancer cell by cDNA microarray].

OBJECTIVE: To study the mechanism of irinotecan-resistant colon cancer by analyzing the differential gene expression patterns with cDNA microarray. METHODS: Total RNA was purified from irinotecan-sensitive colonic cancer cell line, SW480 and its irinotecan-resistance cell line, SW480/CPT. The cRNA retro-transcribed from RNA were labeled with Cy3 fluorescence as probes. The probes were hybridized with Agilent gene chips and the fluorescence images of the chips were obtained with Axon 4000B scanner as well as analyzed with Genepix 3.0 software. The microarray results were confirmed by reverse transcription-polymerase chain reaction. RESULTS: Of the 1598 genes with altered expressions, there were 911 up-regulated genes and 687 down-regulated genes. Glutathione S-Transferase (GST) isoenzyme family GSTA such as GSTA1, GSTA2, GSTA3 and GSTA5 were significantly up-regulated. The expression levels of many Zinc finger protein family members (ZNF) were also differentially regulated. CONCLUSION: GSTA and ZNF subunit genes might play an important regulation role in the irinotecan resistance of colon cancer.

Retroperitoneal neoplasms within the perirenal space in infants and children: differentiation of renal and non-renal origin in enhanced CT images.

PURPOSE: To retrospectively demonstrate the specific CT findings of retroperitoneal neoplasms to diagnosis and differential diagnosis renal and non-renal tumors within the perirenal space in infants and children. MATERIALS AND METHODS: We retrospectively reviewed the clinical data and CT images of 42 consecutive patients with surgically and pathologically proven retroperitoneal neoplasms within the perirenal space. The patients were divided into renal tumors group (n=16) and non-renal tumors group (n=26). The former included nephroblastoma (n=15) and renal lymphoma (n=1), while the latter included neuroblastoma (n=12), retroperitoneal teratoma (n=6), adrenal ganglioneuroma (n=4), retroperitoneal lymphoma (n=2), ectopic pheochromocytoma (n=1) and adrenal cortical carcinoma (n=1). The clinical information of these patients and the major CT imaging findings which were related to lesion localization in the two groups were compared and statistically analyzed using Pearson Chi-Square Test and Risk Estimate. RESULTS: The mean diameter of tumors was 9.82±6.13 cm (n=42 range: 2.3-3 2cm). The demographic data and chief clinical symptoms between the renal tumor group and the non-renal tumor group showed no statistically significant differences (P>0.05). 30.8% (8/26) of non-renal tumor patients presented elevated urinary vanillylmandelic acid (VMA) level, while no patient showed elevated VMA in renal tumor group (P<0.05). Some CT imaging signs of the renal tumors including "crescent sign" (odds ratio, OR=52), "beak sign" (OR=84), "embedded organ sign" (OR=84), and "prominent feeding artery sign" (OR=36) showed significantly higher incidence when compared to the non-renal tumors (P<0.001). The sign of "renal displacement and renal axis rotation" (OR=0.059) was seen in 23 of 26 (88.5%) non-renal tumors, but in only 5 of 16 (31.3%) renal tumors (P<0.001). The sign of "extra-renal central plane of tumor" (OR=0.038) was displayed in 24 of 26 (92.3%) non-renal tumors, but in only 5 of 16 (31.3%) renal tumors (P<0.001). The CT findings such as "pseudocapsule" (OR=38.5), "necrosis and cystic change" (OR=11.2), "vascularity" (OR=16.867), "distant metastasis" (OR=5.96), and "inferior vena cava tumor thrombus" which were thought to be characteristic of renal tumors were observed with significant higher incidence in renal tumors group than in the non-renal tumors group (P<0.05); while CT signs of "irregular mass" (OR=0.045) and "intratumoral calcifications" (OR=0.065) were observed with lower incidence in renal tumors group than in the non-renal tumors group (P<0.05). CONCLUSION: The "crescent sign", "beak sign", "embedded kidney sign" and "renal arteries feeding" are the most specific CT signs suggestive of renal tumors and distinguish them from non-renal origin tumors within the perirenal space. Other CT signs, such as "pseudocapsule", "hypervascular tumors" and "Inferior vena cava tumor thrombus", when present, tumors of renal origin are strongly suggested. On the other hand, CT signs of "irregular mass", "intratumoral calcifications", and associated elevated urinary vanillylmandelic acid strongly suggest the non-renal tumors.

[A clinical study on different decompression methods in cervical spondylosis].

OBJECTIVE: To analyze the different decompression methods to treat cervical spondylosis based on imageological evaluation. METHODS: Two hundred and sixty three consecutive patients with cervical spondylosis between Nov. 2004 and Oct. 2007 were involved in this study. Patients were distributed to different operation groups based on the preoperative imageological evaluation, including anterior or posterior decompression methods. The Anterior method is to use the discectomy of one to three segments, autogenous iliac graft or titanium mesh or cage fusion and titanium plate fixation, or subtotal vertebrectomy of one to two segments autogenous iliac graft or titanium mesh fusion and titanium plate fixation, or discectomy plus subtotal vertebrectomy, The posterior expansive single open door laminoplasty and other operation types. All the patients were divided into different groups by the preoperative imageological evaluation, age, sex and course of diseases. Then we collected each group's preoperative and postoperative JOA scores and mean improvement rate to evaluate the postoperative effect by different decompression methods. RESULTS: Two hundred and thirty five patients were followed up with a mean period of 18 months (range, 4 to 36 months). JOA scores of all patients were improved by different degrees after operations. Anterior and posterior decompression methods both can achieve higher mean improvement rates. There were no significant differences in mean improvement rates between anterior groups, and so did male and female (P > 0.05). The effect will decrease as age increases or the course of disease prolongs. Statistical significance existed among the different age groups and between course groups (P < 0.05). CONCLUSIONS: Anterior and posterior decompression methods both can achieve good effect. The key point is to choose the surgical indication correctly, decompress thoroughly, and make the fusion reliable and fixation firm. In regard to the patients' imageological evaluation, the methods should be differentiated. The anterior operation type included discectomy of one to three segments, subtotal vertebrectomy of one to two segments and discectomy plus subtotal vertebra ectomy.

Effects of n-3 fatty acid, fructose-1,6-diphosphate and glutamine on mucosal cell proliferation and apoptosis of small bowel graft after transplantation in rats.

AIM: To evaluate the effects of n-3 fatty acids (n-3FA), fructose-1,6-diphosphate (FDP) and glutamine (GLN) on mucosal cell proliferation and apoptosis of small bowel graft. METHODS: One hundred and ninety-six inbred strain Wistar rats were grouped as donors and recipients, and underwent heterotopic small bowel transplantation (SBT). n-3FA, FDP and GLN were administered via gastric tube as well as venous infusion for 10 days before and after surgery, respectively. The proliferation and apoptosis of mucosal cells were analyzed with flow cytometry and in situ cell death detection kits. RESULTS: Apparent apoptosis and minor proliferation of mucosal cells of small bowel graft after transplantation were observed. A higher mucosal cell proliferative index and lower apoptotic index were found in all small bowel grafts after supplying with n-3FA, FDP and GLN. CONCLUSION: Nutritional support with n-3FA, FDP and GLN promotes mucosal cell proliferation significantly, and prevents mucosal cell from undergoing apoptosis with different degrees. These regulatory effects on the apoptosis alter the structure and absorption function of transplanted small bowel favorably.

Modified techniques of heterotopic total small intestinal transplantation in rats.

AIM: To establish a successful model of heterotopic total small intestinal transplantation (SIT) in rats in order to reduce the complications and increase the survival rate. METHODS: A total of 196 Wistar rats underwent heterotopic SIT with microsurgical technique. Technical modifications included shortening fasting time and supplying energy before surgery, administering optimal volume of crystalloid fluid to the donor and recipient during surgical procedures, reducing mechanical and ischemic injuries to donor intestine, revascularizing small intestinal graft with a combination of conventional aorta to aorta anastomosis and a cuffed portal vein to left renal vein anastomosis which resulted in an acceptably short warm ischemic time, and also an adequate blood supply and drainage of the graft. RESULTS: The average time for the donor surgery was 86 min +/- 20 min, the mean operative time for the recipient was 115 min +/-20 min and warm ischemia time was shortened to 40 min +/- 5 min. There was a shorter revascularizing time of the graft, the abdominal aorta (AA) to AA anastomosis being 21 min +/- 10 min, and the cuffed portal vein (PV) to the renal vein anastomosis being 5 min +/- 5 min. The one-week survival rate of 98 rats with SIT was 88.78% (87/98), without thrombosis and stenosis of anastomosis. The longest survival time of recipient rats was more than 389 days after SIT, the rats were maintaining normal weight, with perfect intestinal function and intact intestinal histology. CONCLUSION: These modified techniques for SIT would remarkably reduce the complications and improve survival rate in rats, which provided a potentially more consistent and practical model for experimental and clinical studies.