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 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 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 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

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

OBJECTIVETo study the physiopathologic basis of Weikangfu Granule (WKFG) in treating precancerosis of gastric mucosa in patients of chronic gastritis with Pi-deficiency syndrome (CG-PDS).

METHODSOne hundred and fifteen patients of CG-PDS who suffered from intestinal metaplasia (IM) and atypical hyperplasia (ATHP) of gastric mucosa, were divided into two groups. The treated group (n = 61) was treated by WKFG with its ingredients modified according to the syndrome type of patients. The control group (n = 54) was treated with Weishu granule. The histopathological and subcellular ultrastructural changes were detected by optical microscope, screening electronic microscope, transmission electronic microscope and histochemical staining; the nuclear and mitochondrial ultrastructure of gastric mucosa were analyzed with energy dispersion X-ray analyser and image analysis system. And the changes of cAMP, lipid peroxide (LPO), superoxide dismutase (SOD) before and after treatment in the treated group were measured and compared with those of the health control group consisting of 15 volunteers.

RESULTSThe symptomatic and pathological therapeutic effect in the treated group were significantly superior to those in the control group (P < 0.05). The contents of Zn, Cu, cAMP, SOD and (3)H-TdR LCT in gastric mucosa of the treated group before treatment were all lower than those of the healthy control group, yet all these indexes markedly increased after treatment, while serum LPO level, which increased before treatment was lowered after treatment. All the changes showed statistical significance (P < 0.05 or P < 0.01).

CONCLUSIONWKFG can reverse IM and ATHP in patients of CG-PDS, and the effect may be realized by way of increasing the level of Zn, Cu, cAMP and SOD in gastric mucosa, promoting cell differentiation, enhancing cellular immunity and reducing oxygen free radicals and lipid peroxidation.

Adult ; Aged ; Antineoplastic Agents ; pharmacology ; therapeutic use ; Chronic Disease ; Copper ; analysis ; Cyclic AMP ; analysis ; Drugs, Chinese Herbal ; therapeutic use ; Gastric Mucosa ; chemistry ; pathology ; ultrastructure ; Gastritis, Atrophic ; pathology ; Humans ; Lipid Peroxides ; analysis ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Precancerous Conditions ; pathology ; Stomach Neoplasms ; pathology ; Superoxide Dismutase ; analysis ; Syndrome ; Yang Deficiency ; complications ; Zinc ; analysis

OBJECTIVETo test the hypothesis that tissue-engineered cartilage can be bioincorporated with a nonreactive, permanent endoskeletal scaffold.

METHODSChondrocytes obtained from swine articular were seeded onto polyglycolic acids(PGA) scaffold which was incorporated with high-density polyethylene (Medpor). After cultured in vitro for two weeks,the cell-scaffold construct was implanted into subcutaneous pockets on the back of nude mice. Six weeks later,the newly formed cartilage prosthesis was harvested, and a small part of sample was evaluated by gross view, histology, type II collagen immunohistochemistry and biochemistry. PGA scaffold seeded with cells as the control group.

RESULTSThe newly formed cartilage was very similar to normal cartilage in both gross view and histology, and jointed Medpor tightly. The center of control group was hollow.

CONCLUSIONThis pilot technique combining tissue engineering with a permanent success in creating cartilage without "hollow" phenomenon. biocompatible endoskeleton demonstrated

Animals ; Biocompatible Materials ; Cartilage ; transplantation ; Chondrocytes ; cytology ; Materials Testing ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Pilot Projects ; Polyethylenes ; Swine ; Tissue Engineering ; methods ; Tissue Scaffolds

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