Objective: To investigate the distribution of neural cholinesterase and elastic and collagen fibers in rat pancreatic tissues. Methods: Rats were decapitated and the pancreatic tissues were harvested, frozen and made into 8 μm serial sections. A combined-staining method (K-R-VB-P-PA method, a combination of Karnovsky-Roots and Victoria blue-Ponceau-Picric acid) was used for staining and the distribution of neural cholinesterase and elastic and collagen fibers was observed. Results: The results showed that the neural cholinesterase was brown, the elastic fibers were cyan, the collagen fibers were light red, and the smooth muscle was yellow in the pancreatic acinar, islet of Langerhans and blood vessels. Conclusion: K-R-VB-P-PA combined staining method can demonstrate the distribution of neural cholinesterase, elastic and collagen fibers and smooth muscle fibers in the pancreatic tissues synchronously.

Objective: To develop a compound staining method showing collagenic, elastic fibers and melanocyte in skin tissues. Methods: A staining meth od combining Masson Fontana silver nitrate, Ponceau S, Picric acid and Victoria blue (MFSN-P-PA-VB) was used in the present animal skin experimental. Results: The melanin granule was blac k in the skin melanocyte, collagenic fibers were red, the red blood cells at hemorrhagic focus were yellow, and the elastic fibers were green by this MFSN-P-PA-VB staining method. Conclusion: This compound staining method has multicol or and fine contrast, and can better show melanins, the red blood cells at hemorrhagic focus, and collagenic and elastic fibers.

Objective:To observe the effect of Busheng Yiqi Huoxue Capsule on the lipid peroxide, lipofuscin, lipid of aged rats and on the lipid, platelet aggregation and prostaglandin of the experimental hyperlipidemic rats. Methods: Adopting the aged rats and dividing randomly into groups, the indicators of MDA. lipofuscin, lipid were determined; Adopting the hyperlipidemic rats fed by high lipid diet and dividing randomly into groups, the indicators of lipid, platelet aggregation, 6 tone PGF 1? and TXB 2 were determined. Results: Busheng Yiqi Huoxue Capsule can reduce the level of lipid peroxide in the heart, brain, liver and plasma, the level of lipofuscin in the heart and brain and the level of lipid in the aged rats ( P

Objective To identify the diffusion alterations of deep gray matter(GM) and white matter (WM) among Alzheimer's disease (AD), mild cognitive impairment (MCI) and healthy people by atlas?based analysis (ABA), and to investigate the respective relationship with cognitive function. Methods Twenty?one AD patients (AD group), 8 MCI patients (MCI group) and 15 normal controls (control group) were performed by conventional MRI and diffusion tensor imaging (DTI). The raw data of DTI was processed by using DTI studio software to generate the fractional anisotropy (FA) images. Then ABA was used to quantify the FA value in 58 deep GM and WM structures. The differences of FA value among three groups were compared by using one way ANOVA, with a post?hoc analysis. In AD and MCI groups, the partial correlation was further investigated between mini?mental state examination (MMSE) score and FA value in the brain regions that have significant differences between AD and MCI group or between MCI and control group. Results Compared with control group, AD patients showed wide?spread FA decrease in most deep GM and WM regions (corrected P<0.05). The FA values of the hypothalamus, the fornix, the superior longitudinal fasciculus (SLF) and the cingulum in AD group were significantly lower than those in MCI group (corrected P<0.05). The FA value of the right splenium of corpus callosum (SCC) in MCI group was significantly lower than that in control group (MCI:0.550±0.018 vs. Control:0.585±0.026, P<0.05). In AD and MCI group, the FA values of the left hypothalamus, the right hypothalamus, the left cingulum, the right cingulum, and the left SLF were positively correlated with MMSE scores(r=0.502, 0.515, 0.535, 0.527, 0.512; P<0.05). No significant correlation was found between the FA value of the right SCC, the right SLF, the right fornix/stria terminalis, the right fornix and MMSE scores(P>0.05). Conclusion Based on ABA, this study found the diffusion changes not only in the WM but also the deep GM in AD patients, but only WM diffusion disruptions in MCI group. The decreased FA value in the right SCC appeared early, but had no correlation with the cognitive impairment. The FA value in the hypothalamus, the fornix, the SLF and the cingulum decreased with the disease progression, and correlated positively with the cognition decline.

Adenocarcinoma, Clear Cell ; pathology ; Aged ; Female ; Humans ; Kidney Neoplasms ; pathology ; Paranasal Sinus Neoplasms ; secondary

Mitogen-activated protein kinases (MAPKs) are important signaling transduction components well conserved in eukaryotes and play essential roles in various physiological, developmental and hormonal responses in plant. In the present study, a MAPK gene, designated as DoMPK4 (GenBank accession No. JX297597), is identified from a rare endangered medicinal orchid species D. officinale using the reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) methods. The full length cDNA of DoMPK4 is 1 518 bp in length and encoded a 369 aa protein with a molecular weight of 42.42 kD and an isoelectric point of 5.55. DoMPK4 protein contained a serine/threonine protein kinase active site (158-170), a MAP kinase site (71-174), and eight conserved motifs. DoMPK4 had a transmembrane (214-232) but no signal peptide. Multiple sequence alignment showed that DoMPK4 shared high identities (74.9%-80.6%) with MAPK proteins from various plants. Phylogenetic analysis demonstrated that DoMPK4 belonged to group A of the MAPK evolutionary tree, and is closely related to monocots. Real time quantitative PCR (qPCR) analysis revealed that DoMPK4 is differentially expressed among the five organs including leaf, stem, root, seed, and protocorm-like body (PLB). The transcription level of DoMPK4 is the highest in the PLBs with 17.65 fold, followed by seeds, roots, and stems with 5.84, 2.28, and 1.64 fold, respectively. The progressive enhancement of DoMPK4 transcripts in the developing PLBs compared to that in the germinating seeds, suggests a role of DoMPK4 during the development of embryogenic PLBs formation in D. officinale.
Amino Acid Sequence ; DNA, Complementary ; genetics ; DNA, Plant ; genetics ; Dendrobium ; enzymology ; genetics ; Gene Expression Regulation, Plant ; Mitogen-Activated Protein Kinases ; genetics ; metabolism ; Phylogeny ; Plant Leaves ; metabolism ; Plant Proteins ; genetics ; metabolism ; Plant Roots ; metabolism ; Plant Stems ; metabolism ; Seeds ; metabolism ; Sequence Alignment

The zinc-regulated transporters (ZRT), iron-regulated transporter (IRT)-like protein (ZIP) plays an important role in the growth and development of plant. In this study, a full length cDNA of ZIP encoding gene, designed as DoZIP1 (GenBank accession KJ946203), was identified from Dendrobium officinale using RT-PCR and RACE. Bioinformatics analysis showed that DoZIP1 consisted of a 1,056 bp open reading frame (ORF) encoded a 351-aa protein with a molecular weight of 37.57 kDa and an isoelectric point (pI) of 6.09. The deduced DoZIP1 protein contained the conserved ZIP domain, and its secondary structure was composed of 50.71% alpha helix, 11.11% extended strand, 36.18% random coil, and beta turn 1.99%. DoZIP1 protein exhibited a signal peptide and eight transmembrane domains, presumably locating in cell membrane. The amino acid sequence had high homology with ZIP proteins from Arabidopsis, alfalfa and rice. A phylogenetic tree analysis demonstrated that DoZIP1 was closely related to AtZIP10 and OsZIP3, and they were clustered into one clade. Real time quantitative PCR analysis demonstrated that the transcription level of DoZIP1 in D. officinale roots was the highest (4.19 fold higher than that of stems), followed by that of leaves (1.12 fold). Molecular characters of DoZIP1 will be useful for further functional determination of the gene involving in the growth and development of D. officinale.
Amino Acid Sequence ; Cloning, Molecular ; Dendrobium ; chemistry ; classification ; genetics ; metabolism ; Gene Expression Regulation, Plant ; Iron ; metabolism ; Membrane Transport Proteins ; chemistry ; genetics ; metabolism ; Molecular Sequence Data ; Phylogeny ; Plant Proteins ; chemistry ; genetics ; metabolism ; Plants ; chemistry ; classification ; genetics ; Sequence Alignment ; Zinc ; metabolism

Objective To analyze clinical features and sum up experience for the treatment of ischemic bowel disease. Methods Clinical data of 73 patients with the diagnosis of ischemic bowel disease were retrospectively analyzed. ResultsTwenty-eight patients were male and 45 patients were female. The median of age was 65 years (range of 38 to 89 years). Forty-eight patients were associated with hypertension, 23%(17/73) patients had a history of coronary disease and 15% (11/73) had diabetes. Seventy patients presented symptom of abdominal pain and 93% (68/73) had hematochezia. Symptoms relieved by conservative treatment in 96% (63/66) patients. Nine patients underwent a surgery. One patient died of sepsis postoperatively. One suffered from colostomy necrosis and leakage of the rectum segment. Conclusion 1. Elder patients presenting symptoms of abdominal pain and hematochezia, especially with a history of cardio-cerebrovascular disease and diabetes should be considered for the possibility of ischemic bowel disease. 2. Most patients with ischemic bowel disease could be successfully treated by conservative therapy. 3. Surgery for patients with chronic relapsing and nonresponsible symptoms was difficult and patients often suffer from high postoperative complications.

Objective To investigate the effect of early enteral nutrition combined with probiotics on the nutritional status of patients with severe head injury. Methods Thirty-five patients with severe head injury were randomized into early enteral nutrition group ( control group, n = 18 ) and early enteral nutrition combined with probiotics group (observational group, n = 17). All the patients received enteral nutrition via nasogastric tube within 24-48 hours following admission. Hemoglobin, lymphocyte count,plasma concentrations of total protein, albumin, prealbumin, cholinesterase, and fasting blood glucose were determined before enteral nutrition and at days 8, 15, 21 after enteral nutrition. Arm circumference (AC), triceps skinfold thickness (TSF) and ann muscle circumference ( AMC ) were measured at the same time points. Gastrointestinal reactions as well as the time to first defecation were recorded. Glasgow coma scale (GCS) was assessed. Results There was no difference in terms of hemoglobin, lymphocyte count, plasma concentrations of total protein, prealbumin, and GCS score at each time point between the two groups ( P ＞ 0. 05 ). At day 8 after enteral nutrition, the patients in the observational group exhibited significantly lower level of fasting blood glucose than those in the control group (P ＜ 0.05 ). At day 15,the concentration of plasma cholinesterase in the observational group was significantly higher than that in the control group ( P ＜ 0. 05 ). At day 21, the observational group showed a tendency in increase of plasma albumin level in comparison with the control group( P = 0. 054), and the decrease of AC and AMC was obviously lower than that in the control group ( P ＜0. 05 ) compared with pre-enteral nutrition. The observational group showed less gastro-oesophageal reflux and constipation as well as earlier time to first defecation compared with the control group ( P ＜ 0. 05,0. 01 ). Conclusion Compared with the ordinary enteral nutrition, the enteral nutrition combined with probiotics can attenuate metabolic disorders and improve the nutritional status of the patients with severe head injury.

Aim To establish in vitro blood-brain barrier (BBB) model with characteristics of simulation of in vivo BBB by primi-tive co-culture of brain-microvessel endothelial cells (BMECs) with brain-microvessel pericytes (BMPC)and astrocytes (AS). Methods BMECs,BMPC and AS from SD rats were primitively isolated,purified and cultured,and then primitive culture cells were identified by cellular morphological and immunocytochemi-cal staining methods.Five types of in vitro BBB models were es-tablished by using Millicell culture insert (pore diameter 0.4μm)and their barrier functions were evaluated by detection of transendothelial electrical resistance (TEER),permeability of sodium fluorescent (Na-FLU ),expression of alkaline phospha-tase (AKP)and γ-glutamyl transpeptidase (γ-GT1 ),and simi-larity of permeation amount for positive drugs in vitro and in vivo BBB conditions.Results Primitive culture of BMECs presented typical pebbles-like structure,BMPC presented larger soma with branching property,AS presented slender synapse and shallower cytoplasm.Moreover,immunocytochemical staining results iden-tified primitive cells were targeted cells.TEER value for co-cul-ture of BMECs,BMPC and AS reached (478 ±25 )Ω· cm2 , permeability coefficients (Papp )value of Na-FLU was [(8.23 ± 0.78) ×10 -6 ]cm·s-1 ,expression of AKP and γ-GT1 were (6.90 ±0.27 )King unit · g-1 Pro and (4.39 ±0.32 )μg · g-1 Pro respectively.Moreover,good correlation could be found in Papp for positive controls in vitro and in vivo BBB models (R2=0.92).Conclusion The established in vitro BBB model by using primitive co-culture of BMECs with BMPC and AS posses-ses in vivo BBB properties in cell morphology,structures and barrier functions,and can be used as a powerful tool for studying physiology,pathology of BBB and screening candidate com-pounds.