Objective:To clone and express panallergen profilin from the pollen of coco(Cocos nucifera Linnaeus).Methods:RT-PCR and RACE methods were applied to clone the full-length panallergen genes from coco pollen and the sequence was analyzed.The specific primers were designed.The ORF of profilin of coco pollen was amplified with RT-PCR and cloned into the expression vector pET 28a.Expression of the recombinant coco pollen profilin was carried out in E.coli BL21(DE3) and the purification of the recombinant protein was performed via affinity chromatography with Ni2+ coupled to sepharose.IgE reactivity to recombinant coco pollen profilin was investigated by immunoblot.Results:The complete sequence of coco pollen profilin was cloned.The sequence was 608 bp and included an open reading frame(396 bp) coding for 131 amino acids.Sequence analysis showed that the deduced protein was an acidic protein with an estimated molecular mass of 14.19 kD and a pI of 4.61.The GeneBank accession number of the clones was EF173598.After overexpressed in E.coli BL21(DE3),the recombinant protein was purified through affinity chromatography with Ni2+ coupled to sepharose.Immunoassay showed that the recombinant allergen has good IgE binding capacity.Conclusion:The profilin of coco pollen is expressed successfully in BL21(DE3),which will be used as a base for further study on coco pollen related allergy.

Objective To explore the pulmonary function,DVH and radiation pneumonitis after three-dimensional conformal radiation treatment ofⅢphase non-small-cell lung cancer.Methods 71 pa- tients (male 52,female 19,median age 63,KPS≥80) were evaluated by pulmonary function tests before radiotherapy and in M1 and M3 after radiotherapy respectively.After 3 months of follow-up time,it reviewed the appearance and grade of radiation pneumonitis.Then V_(20),V_(30) and MLD were worked out from dose vol- ume histogram.Results All patients completed radiotherapy,and total dose was 66-70 Gy.FVC (L), FEV1 (L) and CLCO were (2.58?0.65) L,(1.85?0.58) L and (15.15?4.65)ml/(min)before radio- therapy,with (2.96?0.76) L,(2.13?0.65) L and (14.71?3.92) ml/(min) in Ml after radiotherapy, with (2.65?0.61) L,(1.92?0.52) L and (13.15?3.71)ml/(min)in M3 after radiotherapy.The ac- cidence of radiation pneumonitis was 30%,moderate and severe radiation pneumonitis was 7%.With V_(20), V_(30) and MLD increasing,the grade of radiation pneumonitis was increasing.V_(20),V_(30) and MLD were related to the change in CLCO value among before,M1 and M3 after radiotherapy,and the correlation coefficient was more than 0.2.Conclusions There is a relationship in the pulmonary function,DVH and radiation pneu- monitis surely.The change in C_LCO value between before radiotherapy and M1 after radiotherapy could pre- dict the radiation pneumonitis.V_(20),V_(30) and MLD are not only correlated to radiation pneumonitis evidently but the change in FEV_1 and C_LCO after radiotherapy.

Objective To study the susceptible factors of atherosclerotic stenosis before the coronary artery myocardial bridges.Methods The data from 88 myocardial bridge cases which received coronary angiography were statistically analyzed.Sixty-seven cages which suffered from atherosclerofic stenesis in proximal of the myocardial bridges were recruited into group A,and the other 21 cases which suffered no atherosclerotic stenosis or from atherosclerotic stenosis in distal of the myocardial bridges were mcmited into group B.Difference of the age,gender,length of myocardial bridge,systolic blood pressure (SBP),diastolic blood pressure (DBP),pulse pressure (PP),the oppression degree of myocardial bridge (Nobel classification),fasting plasma glucose (FPG),and blood fat,and so on,in two groups,were observed and statistically analyzed.Results The difference of the Nobel classification,SBP and PP in two groups showed a statistical significance (P＜0.05).While the difference of the age,gender,length of myocardial bridge,DBP,FPG,total cholesterol,low-density hpoprotein in two groups showed no statistical significance (P＞0.05).A further regression analysis suggested that Nobel classification and PP had a correlation with the comphcation of stonosis before the myocardial bridge (r=3.0569,0.9740,P＜0.05).Conclusions High blood pressure cases are liable to suffer from myocardial bridge.Myocardial bridges themselves trend to promote or accelerate the atherosclerotic stenosis of the coronary arteries before.them.The oppression degree of myocardial bridge and PP has a correlation with the complication of stenosis before the myocardial bridge,while has no correlations with age,gender,bloodfat,SBP,DBP,FPG,length of myocardial bridge,and so on.

Objective To analyze CT findings correlated with pathologic findings in ectopic pancreas of the stomach.Methods CT scans of 15 surgically proven cases of ectopic pancreas of the stomach were reviewed,and enhanced CT scan was performed in 11 cases.CT findings were correlated with the pathologic findings.Results All cases had single lesion,and all lesions showed homogeneous density on plain scans without cystic or malignant changes.The size ranged from 1.3 to 3.1 cm,with mean diameter of (1.9? 0.2)cm.The lesions were round or oval in shape with broad base against the gastric wall.Two showed central umbilication sign.Only 2 cases were correctly diagnosed prior to operation and the rest were misdiagnosed or diagnosed indistinctly.The locations were in the gastric antrum in 11 cases,in the body in 3,and in fundus in one;The ectopic pancreas located in the greater curvature in 10,and in the lesser curvature in 5.Homogeneous or inhomogeneous strong enhancement similar to the pancreas was seen in 8 cases and they consisted mainly of pancreatic acini with the same histologic features as the pancreas.Three cases showed poor enhancement and consisted mainly of ducts and hypertrophied muscle,pancreatic acini were a minor component.Conclusion Ectopic pancreas of the stomach showed characteristic locations with the findings of snhmucosal diseases.Different enhancing patterns were correlated with their pathologic findings.

cDNA for Insulin-like growth factor binding protein 3 was cloned and constructed a prokaryotic expression vector--pET-DsBA-IGFBP3. The construct was transformed into E. coli BL21 (DE3)plysS. The induced fusion protein (D-IGFBP3) was expressed successfully in soluble form. We obtained D-IGFBP3 the purify of which is over 95% after purification by His affinity chromatography. The product was identified by Western-blot. The cell assay showed that the obtained fusion protein can inhibit the growth of MCF-7 and bind with IGF-I in vitro.
Blotting, Western ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Chromatography, Affinity ; Dose-Response Relationship, Drug ; Electrophoresis, Polyacrylamide Gel ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli ; genetics ; Gene Expression ; Humans ; Insulin-Like Growth Factor Binding Protein 3 ; genetics ; metabolism ; pharmacology ; Insulin-Like Growth Factor I ; metabolism ; Protein Binding ; Recombinant Proteins ; isolation & purification ; metabolism ; pharmacology ; Solubility

BACKGROUNDRetinal pigment epithelial (RPE) cells play an important role in the occurrence of choroidal neovascularization (CNV). Vascular endothelial growth factor (VEGF) as a positive regulatory growth factor is produced by the RPE in an autocrine or paracrine manner, promoting CNV development. Duplexes of 21 nt RNAs, known as short interfering RNAs (siRNAs), efficiently inhibit gene expression by RNA interference when introduced into mammalian cells. We searched for an efficient siRNA to interfere with VEGF expression in RPE cells and shed light on the treatment of CNV.

METHODSHuman primary RPE (hRPE) cells were cultured and identified. Three pairs of siRNAs were designed according to the sequence of VEGF 1-5 extrons and synthesized by T7 RNA polymerase transcription in vitro. To evaluate the inhibitory activity of T7-siRNAs, hRPE cells were transfected via siPORT Amine. The interfering effect of T7-siRNAs in hRPE cells was examined by semiquantitative reverse transcription-polymerase chain reaction and immunofluorescence.

RESULTSThree pairs of T7-siRNAs synthesized by in vitro transcription with T7 RNA polymerase suppressed VEGF gene expression with efficiency from 65% to 90%. T7-siRNA (B), targeted region at 207 nt to 228 nt and double stranded for 21 nt with 2 nt UU 3' overhangs, was the most effective sequence tested for inhibition of VEGF expression in hRPE cells. Compared with nontransfected cells, the mean fluorescence in hRPE cells transfected with T7-sRNAs was significantly less (P < 0.01). siRNA with a single-base mismatch and ssRNA(+) did not show suppressing effect. Furthermore, it was found that siRNAs had a dose dependent inhibitory effect (5 to 10 pmol).

CONCLUSIONT7-siRNA can effectively and specifically suppress VEGF expression in hRPE cells and may be a new way to treat CNV.

Base Sequence ; Cells, Cultured ; Choroidal Neovascularization ; therapy ; DNA-Directed RNA Polymerases ; metabolism ; Humans ; Molecular Sequence Data ; Pigment Epithelium of Eye ; cytology ; metabolism ; RNA Interference ; RNA, Small Interfering ; biosynthesis ; pharmacology ; Transcription, Genetic ; Vascular Endothelial Growth Factor A ; antagonists & inhibitors ; genetics ; Viral Proteins ; metabolism

BACKGROUNDMost of the studies on traditional Chinese medicine (TCM) 'spleen' deficiency syndrome in the recent 30 years were conducted only on the basis of single functional index, neglecting the study on the pathophysiologic internal relationship between spleen deficiency syndrome and gastric diseases in modern medicine. But it was at the subcellular molecular biological level that we explored the pathophysiologic basis of classification of spleen deficiency in chronic gastritis by detecting the bioactive substances in gastric mucosa nuclei and mitochondria.

METHODSBy means of optical microscope, scanning electron microscope (SEM), transmission electron microscopy (TEM) and histochemical staining, we conducted histopathological, subcellular ultrastructural analysis and nuclei and mitochondrial ultrastructural analysis of gastric mucosa of 188 spleen deficiency patients and of 42 voluntary blood donors. At the same time, bioactive substances were measured by means of X-ray energy dispersive analysis system (EDAX) image analysis system, radioimmunoassay method and chemiluminescence method.

RESULTSThe content of cAMP, superoxide dismutase (SOD), Zn and Cu in gastric mucosa, and the content of Zn and Cu in mitochondria decreased progressively in order of groups: healthy control (HC), spleen Qi deficiency without organic lesion (F-SQD), spleen Yang deficiency without organic lesion (F-SyangD), disease without symptoms group, spleen Qi deficiency with organic lesion (G-SQD), spleen Yang deficiency with organic lesion (G-SyangD), spleen Yin deficiency (SyinD) and spleen deficiency with Qi stagnation (SDQS), chronic spleen deficiency gastritis (CSG) and chronic atrophic gastritis (CAG); decreased in order of HC, intestinal metaplasia (IM)Ia, IMIb, IMIIa and IMIIb, P < 0.05. The content of DNA, Zn and Cu in nuclei progressively increased in order mentioned above, P < 0.05.

CONCLUSIONSThe quantitative changes of gastric mucosal cAMP, SOD, Zn, Cu, of mitochondrial Zn, Cu and of nuclear DNA, Zn and Cu are not only the substance base on which the lesion of gastric mucosa tissue structure occurs, but also the substance base on which spleen deficiency is classified. G-SQD and G-SyangD were more likely to be found in low-grade or middle-grade CSG and CAG, while SyinD and SDQS in middle-grade or high-grade CSG, CAG and IMIIb.

Adult ; Aged ; Chronic Disease ; Cyclic AMP ; analysis ; Female ; Gastric Mucosa ; pathology ; ultrastructure ; Gastritis ; metabolism ; pathology ; Humans ; Lipid Peroxides ; blood ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Splenic Diseases ; classification ; Superoxide Dismutase ; analysis

BACKGROUNDMagnetic resonance imaging (MRI) is prone to be deformed by artifacts caused by the presence of metallic materials. The aim of this study was to evaluate the artifacts from galvano-ceramic and metal-ceramic crowns in MRI, in order to analyze their influences on diagnostic interpretation of MRI.

METHODSGalvano-ceramic and metal-ceramic crowns (Bio98, Wiron99, SP-78, BioKC97) were fabricated with the same model. All materials were imaged by means of 1.5T MRI apparatus with three different sequences, T1-weighted spin-echo (T1-weighted SE), T2-weighted spin-echo (T2-weighted SE) and Gradient echo (GE). Mean and standard deviation of distilled water signal intensity (SI) around the sample in the region of interest (500 mm2) enclosing the whole artifacts were determined, and compared for evaluation of the homogeneity of signal intensity. Images around the sample were acquired and evaluated.

RESULTSThere were statistically significant differences in the values of signal intensity between acrylic resin control and BioKC97, Wiron99 in the three sequences (P<0.001). The mean values of signal intensity for Bio98, SP-78 were significantly different from that of acrylic resin control (RE) in GE sequence (P<0.001). No difference was showed between acrylic resin control and galvano-ceramic crown (P>0.05). Images showed that the greatest artifact was a 25 mm ring with distortion in Wiron99 in GE sequence.

CONCLUSIONSThis in vitro study suggested that galvano-ceramic crown had no influence on the MRI, while metal-ceramic crowns caused moderate artifacts in the MRI. Therefore, galvano-ceramic restoration is a valuable alternative in dentistry.

Artifacts ; Ceramics ; Crowns ; Dental Alloys ; Humans ; Magnetic Resonance Imaging ; methods ; Metals

BACKGROUNDSpleen in Traditional Chinese Medicine (TCM) is not actually the spleen in the anatomic sense designated in western medicine because its functions basically belong to the physiological category of digestive system in modern medicine, and it represents a macroscopic concept of digestion, absorption and nutrition metabolism. Spleen deficiency syndrome refers to the clinical phenomena such as hypofunction of digestion, absorption and nutrition metabolism. By integrating TCM with modern medicine, this paper is intended to explore the pathological basis of classification of spleen deficiency in chronic gastritis.

METHODBy means of optical microscope, scanning electron microscope (SEM), transmission electron microscope (TEM) and histochemical staining, we conducted histopathological and subcellular ultrastructural (nuclei and mitochondrial) analysis of gastric mucosa of 188 patients of spleen deficiency, and that of 42 voluntary blood donors without clinical symptoms.

RESULTSThe gastric mucosa of patients with spleen Qi deficiency (SQD) and spleen yang deficiency (SyangD) could either be affected by organic lesion (type G-occurring on the basis of chronic superficial gastritis (CSG), chronic atrophic gastritis (CAG)) or unaffected (type F-chiefly belonging to functional indigestion); spleen yin deficiency (SyinD) and spleen deficiency with Qi stagnation (SDQS) both occurred on the basis of CSG and CAG; and the degree of mucosa inflammatory cells infiltration, the degree of decrease in glands propria, and the incidence of IMIIb in CSG and CAG were more serious than those of G-SQD and G-SyangD, P < 0.05 - 0.01.

CONCLUSIONSpleen deficiency syndrome is likely to occur on the basis of organic lesion of gastric mucosa (disease with symptoms of both CSG or CAG and spleen deficiency symptoms), as well as on the basis of inorganic lesion of gastric mucosa (nondisease with symptoms, which is, despite spleen deficiency symptoms, there is no CSG or CAG). Besides, the clinical phenomenon of disease without symptoms (despite CSG or CAG, there is no spleen deficiency symptoms) occurres because of such factors as genetic diathesis and compensation. The lesion degree of CSG or CAG and the incidence of IMIIb of SyinD and SDQS are more serious than those CSG and CAG of G-SQD and G-SyangD.

Adult ; Chronic Disease ; Digestive System Diseases ; classification ; Epithelial Cells ; ultrastructure ; Female ; Gastric Mucosa ; pathology ; ultrastructure ; Gastritis ; pathology ; Humans ; Male ; Medicine, Chinese Traditional ; Splenic Diseases ; classification

OBJECTIVETo analyze the clinical characteristics, influencing factors and outcome of recurrent patients with early stage bulky cervical carcinoma.

METHODSBetween January 1(st) 2000 and December 31(st) 2009, 76 patients with stage Ib2 and IIa2 bulky cervical carcinoma developed recurrence and (or) metastasis. The recurrence time, recurrence location, recurrence-related factors, treatment and survival were analyzed.

RESULTSThe median follow up was 44 months (9-137 months). The overall recurrence and (or) metastasis rate was 22.6%. The 1-, 1-2, 3-5 and 5-year recurrence and (or) metastasis rates were 38.2%, 27.6%, 30.3% and 3.9%, respectively. The 5-year survival rate of local recurrence was 34.5%, that of distant metastasis was 23.6%, and that of distant metastasis with synchronous pelvic recurrence was 11.1%, (P = 0.555). The 5-year survival rate of patients who received surgery plus chemotherapy, radiation plus chemotherapy and chemotherapy alone after recurrence and (or) metastasis were 53.3%, 30.7% and 24.6%, respectively (P = 0.686). Univariate analysis demonstrated that tumor recurrence and (or) metastasis in patients of the stage Ib2 and IIa2 bulky cervical carcinoma were influenced by the disease stage, pelvic lymph node metastasis, deep cervical stromal invasion, lymphovascular tumor thrombus and pathological types. Multivariate regression analysis demonstrated that pelvic lymph node metastasis, lymphovascular tumor thrombus and pathological types were the key factors affecting the recurrence and (or) metastases of the stage Ib2 and IIa2 bulky cervical carcinoma. Subgroup analysis showed that pelvic lymph node metastasis and stage were the main factors affecting the local recurrence in those patients, and the pathological type, vascular tumor thrombus and pelvic lymph node metastasis were the main factors affecting the distant metastasis.

CONCLUSIONSRecurrence and(or) metastasis of early stage bulky cervical cancer are mostly happened within 2 years post operation. Patients with pelvic lymph node metastasis have high probability to develop local recurrence and distant metastasis. Patients with non-squamous cell carcinoma and lymphovascular tumor thrombus are more likely to develop distant metastasis. Neoadjuvant chemotherapy does not decrease local recurrence and distant metastasis in patients with stage Ib2 and IIa2 bulky cervical carcinoma. Individualized treatment is advised for recurrent patients.

Adenocarcinoma ; pathology ; secondary ; surgery ; therapy ; Carcinoma, Squamous Cell ; pathology ; secondary ; surgery ; therapy ; Chemotherapy, Adjuvant ; Female ; Follow-Up Studies ; Humans ; Hysterectomy ; Lung Neoplasms ; drug therapy ; secondary ; Lymph Node Excision ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Multivariate Analysis ; Neoadjuvant Therapy ; Neoplasm Invasiveness ; Neoplasm Recurrence, Local ; surgery ; therapy ; Neoplasm Staging ; Pelvis ; Radiotherapy, Adjuvant ; Survival Rate ; Tumor Burden ; Uterine Cervical Neoplasms ; pathology ; surgery ; therapy