As a member of tyrosine kinases(TK) family,Epidermal growth factor receptor(EGFR) has an activity of intrinsic protein tyrosine kinase,which plays an essential role in the regulation of signal transduction in the cells.Due to the abnormal expression of EGFR-TK,the certain type cancers may developed and progressed.Based on that,the inhibitors of EGFR-TK could be effective medicines for the treatment of cancer.The EGFR-TK domain was amplified by RT-PCR with RNA of HUVCEs cells as the template and expressed in E.coliBL21(DE3) using plasmid pET30a as vector.The recombinant protein was purified with the affinity chromatography(Ni-NTA),which was identified to have kinase activity catalyzing the substrate phosphorylated with ATP in the enzymatic reaction.Using the recombinant EGFR-TK as target,the screening model for enzymatic inhibitors was constructed.

Objective To study the clinical significance of P - selection expression in children with viral encephalitis and the correlation between this expression and the cerebral infarction with critical viral encephalitis. Methods Flow cytometric was employed to detect the expression of P- selection on the surface of platelet membrane in 44 children with viral encephalitis(20 light patients and 24 critical patients) and 20 healthy control children. The area of the cerebral infarction was determined by computed tomographic scan in 20 patients with critical viral encephalitis. The correlation between the two variables was analyzed. Results The expressions of P - selection on the surface of platelet membrane on less than 5 days and on 2 weeks after the onset of viral encephalitis were significantly higher in critical patients than those in normal control children and light patients( P

Objective To observe the antihypertensive effect of indapamide combined with enalapril on spontaneously hypertensive rats(SHRs).Methods Forty SHRs were randomly divided into 4 groups: control,indapamide,enalapril,and indapamide+enalapril(n=10 in each group).Medicine in varied doses was given to rats by intragastric administration.Variations of weight,heart rate and blood pressure were measured. Results Varied doses of medicine did not exert significant effects to the weight and heart rate of SHRs during and after the administration.In indapamide+enalapril group,the pressure of SHRs was significantly decreased with varied doses compared to that before the administration(P

Animals ; Arsenic ; toxicity ; Comet Assay ; DNA Damage ; drug effects ; Lymphocytes ; drug effects ; metabolism ; Male ; Malondialdehyde ; metabolism ; Rats ; Rats, Wistar ; Tobacco Smoke Pollution ; adverse effects

Objective To know the antitumor effect and value of vinorelbine in chemotherapy with relapsed breast cancer.Methods To receive breast cancer tissue from primary and relapsed patients by surgery,then to obtain mammary cancer cells by collagenase.The antitumor effect of 5 chemotherapy drugs with breast cancer cells was measured by culture in vitro and MTT.Results With primary cancer cells from primary breast cancer,the antitumor effect of paclitaxel and docetaxel(sensitivity is 91.04%,92.54% respec- tively) is better than adriamycin,epirubicin and gemcitabine(sensitivity is 73.13%,74.63%,71.64% re- spectively;P

BackgroundPterygium is an ocular surface disease of abnormal cell proliferative kind and angiogenesis plays an important role in its development and recurrence.Several anti-angiogenic therapies have been used to treat pterygium,but there very few studtes for the in vivo observation of the microvessles in pterygium.ObjectiveThis study was to observe angiogenesis in pterygium with a high-resolution confocal microscope in vivo and to perform immunohistochemical study in vitro.MethodsA prospective case-controlled study was designed.Twenty eyes of 20 consecutive patients with primary pterygia and 20 age- and sex-matched patients with inner eye diseases and strabismus with normal conjunctiva were enrolled in this study.An in vivo confocal microscopy imaging system (Heidelberg Retina Tomograph Ⅱ Rostock Cornea Module) was used to collect microvascular pictures from the anterior part of pterygia and normal nasal conjunctiva of controls,and then immunochemistry was performed to examine the expression of CD31 in microvessel in vitro.The vascular density values were compared between these two groups.The correlation of vascular density values between in vivo Heidelberg Retina Tomograph and in vitro immunohistochemistry was calculated.Written informed consent was obtained from pationts before any examination and surgery.ResultsUnder the in vivo confocal microscope,the microvessel density was (8929±2993) μm/mm2 and (4202 ±692)μm/mm2,respectively,in pterygium and the normal conjunctiva group with a statistically significant difference between them (t =6.881,P＜0.01 ).Immunochemistry revealed that the expression of CD31 to measure vascular density was ( 21.00 ± 4.06/400 × field ) and ( 6.07 ± 1.75/400 × field ) in pterygium and the normal conjunctiva group,showing significant difference (t =12.312,P＜0.01 ).Positive correlations were found in the vascular density values between in vivo corneal laser confocal microscopy examination and in vitro immunochemistry examination in both the pterygium group and normal conjunctiva group (pterygium group:r=0.649,P＜0.01 ;normal conjunctiva group: r=0.572,P＜0.01 ) ConclusionsIn vivo confocal microscopy imaging is superior to in vitro immunochemistry in evaluating the microvessel of pterygium.The results of this study offer a new way index for further investigation of the biological behavior of pterygium and its mechanism.

Objective To observe clinical effects of Baixuan xiatare capsules combined with Baskin and compound Flumetasone on elderly patients with chronic eczema .Methods 160 elderly patients with chronic eczema were randomly divided into observation group and control group ,80 cases in each group , the observation group with XTR combined with Baskin , compound Flumetasone treatment , control group using Baskin combined with compound Flumetasone treatment for 4 weeks,the clinical efficacy was compared between the two groups .Results In the obser-vation group,the effective rate was 94.7%,the effective rate of control group was 82.1%,two groups have statistical significance with the efficiency difference (χ2 =5.862,P<0.05).Conclusion The XTR combined with Baskin and compound Flumetasone have better clinical effects than Baskin combined with compound flumetasone treatment with elderly patients with chronic eczema .

Objective To analyze the causes of death and prognostic indicators of rheumatoid arthritis (RA).Methods A retrospective investigation was performed on 91 RA patients who were admitted into our hospital from 1990 to 2011.The clinical features and laboratory data were studied to assess the causes of death and the relationship between causes of death and disease activity and treatment.T test and x2 test were used for statistical analysis.Prognostic indicators of mortality were studied by Cox propor-tional hazards models.The prognostic indicators of RA were also analyzed.Results The three most common causes of death in patients with RA were infection (57％),cardiac-cerebral vascular diseases (13％) and cancer (12％).Severe disease activity and presence of extra-articular manifestations were more common among the patients who died.Seventy-five percent patients presented with systemic symptoms including fever,fatigue and weight loss,followed by pulmonary fibrosis (32％) and pleural effusion (27％).Cox regression analysis revealed that the presence of extra-articular features was independent risk determinant for mortality while DMARDs treatment was the independent protect determinant.Conclusion Infection,cardiac-cerebral vascular diseases and cancer are the main causes of death in RA.The presence of extra-articular features is the independent risk factor of RA.

Pericellular matrix (PCM) is a narrow tissue region surrounding chondrocytes, which "chondron" with its enclosed cells. A number of studies suggested that PCM is rich in proteoglycans, collagen and fibronectin, and plays an important role in regulating microenvironment of chondrocytes. Direct measures of PCM properties through micropipette aspiration technique showed that PCM was different from mechanical property of chondrocytes and nature extracellular matrix. However, the function of PCM is not clear, and need further study.
Animals ; Biomechanical Phenomena ; Chondrocytes ; chemistry ; cytology ; metabolism ; Extracellular Matrix ; chemistry ; metabolism ; Humans

Abstract: Objective To establish the duplex TaqMan RT-PCR method for detection of Entamoeba histolytica and Giardia lamblia in fecal samples. Methods Primer pairs and probes for Entamoeba histolytica and Giardia lamblia were designed and duplex TaqMan RT-PCR amplification system was constructed. PCR products were inserted into the pUC57 plasmid, and the lower limit of detection of the method was determined. Clinical stool samples were tested in order to evaluated the efficacy of the method. Results The detection limits of duplex TaqMan RT-PCR were 31.6 copies/μL for Entamoeba histolytica and 32 copies/μL for Giardia lamblia, respectively. Of the total of 212 clinical stool samples tested, all 3 samples with E. histolytica-positive patients by microscopy were positive by PCR, while 1 from 209 samples with E. histolytica-negative patients by microscopy were positive by PCR, and the remaining samples were negative. For Giardia lamblia, all 8 samples positive by microscopy were positive by PCR, and 1 from 204 sample with a microscopy-negative patient was positive by PCR, and the remaining samples were negative. The amplification product sequencing and blast analysis were used to confirm that the amplified sequence in the specimen of a patient with negative microscopy but positive PCR belongs to the targeted pathogen, supported by clinical symptoms and laboratory test results. PCR results for other diarrhea-causing pathogens were negative, indicating no cross-reactivity. Conclusions The dual TaqMan RT-PCR method developed in this study can not only detect microscopy-positive samples of Entamoeba histolytica and Giardia lamblia but also can detect samples that were missed by microscopy, with higher sensitivity than the microscopy method. Further, this detection method does not cross-react with other diarrhea-causing pathogens, including cross-react with other diarrhea-causing pathogens including Iodamoeba butschlii, Blastocystis hominis, Plesiomonas, Aeromonas, Salmonella, Shigella, Sphaerozoum fuscum, and Entamoeba hartmani, thus has a good specificity.