Objective Making the fusion protein of IgG-binding peptide with enhanced green fluorescent protein(EGFP) and determining its bioactivity.Methods The enhanced green fluorescent protein(EGFP) gene was cloned into pEZZ 18 vector containing ZZ peptide gene to construct expression vector pSpA-EGFP-His.The fusion protein was expressed in E.coliDH5? and its bioactivity was detected by competitive ELISA and fluorescence properties.Results The fusion protein migrated at approximately 42kD in SDS-PAGE,which correspond to the theoretical molecular weight.The spectra of SpA-EGFP fusion protein was similar to what was reported.SpA-EGFP competed with SpA-Peroxidase to bind IgG.Conclusion The plasmid pSpA-EGFP-His correctly expressed in E.coli.The fusion protein retains the bifunctional effects of EGFP and IgG-binding activity.

Objective:To investigate the effects of arecoline on the expression of human telomerase reverse transcriptase(hTERT)mRNA and protein in cultured normal human oral mucosa keratinocytes(KCs),and then to investigate the role of hTERT in carcinogenesis of oral submucous fibrosis(OSF).Methods:Normal human oral mucosa KCs were cultured in vitro,and in experiment group,KCs was divided into 0.03,0.06,0.09 g/L arecoline group,and 0.0 g/L arecoline group served as the control.The hTERT mRNA and protein expression of KC was examined by reverse transcription polymerase chain reaction(RT-PCR)and Western blotting.Results:Arecoline induced the hTERT mRNA and protein expression of KC in a dose dependent manner.hTERT mRNA and protein expression of KC were increased by 0.03,0.06 and 0.09 g/L arecoline,when compared with the control group(P

Objective:To explore the effects of myofibroblast in the pathogenesis of oral submucous fibrosis(OSF).Methods:The expressions of ?-SMA in the tissues of OSF and normal control were detected by immunohistochemistry. Fibroblasts were isolated from oral mucosa of OSF patients and healthy controls and were cultured under stimulation by arecoline.The expressions of ?-SMA in OSF fibroblasts and normal fibroblast were detected by immunocellchemistry and RT-PCR.Results:(1)No positive expression was found in normal tissues except in the blood vessel,but in OSF tissue,?-SMA positive expression was found in many spindle cells in the oral submucousal layer.(2)Few fibroblasts in vitro from normal controls were ?-SMA positive.(3)Most of the second passage of OSF fibroblasts were ?-SMA positive(85.80%?3.56%). (4)The expression of ?-SMA in fibroblasts could not be increased by the stimulation of arecoline.Conclusion:Myofibroblasts may play an important role in the pathogenesis of OSF,and the appearrence of myofibroblasts in OSF may not be directly induced by the stimulation of arecoline.

Objective:In order to elucidate the derivation and differentiation mechanisms for derivation of myofibroblasts in oral submucous fibrosis.Methods:Oral keratinocytes and fibroblasts were isolated and cultured,and a co-cultur system composed of the keratinocytes and fibroblasts was established in vitro.Expressions of ?-SMA protein and mRNA in fibroblasts were detected by immunohistochemistry and RT-PCR. The level of TGF-?1 in conditioned medium were evaluated by ELISA.Results:The expressions of ?-SMA mRNA and protein in fibroblasts co-cultured with keratinocytes preprocessed by arecoline was higher than that in fibroblasts co-cultured with keratinocytes and without preprocessed by arecoline. The level of TGF-?1 in conditioned medium of fibroblasts-keratinocytes preprocessed by arecoline co-cultures was higher than those of fibroblasts cultured alone.Conclusion:The derivation of myofibroblast from FB can be promoted by keratinocytes preprocessed by arecoline and TGF-?1 may play a role in the process.

Objective:To investigate the role of human telomerase reverse transcriptase(hTERT) in carcinogenesis of oral submucous fibrosis(OSF).Methods:The expression of hTERT was examined by immunohistochemical SP method in 10 cases of normal oral mucosa, 46 cases of OSF,14 of carcinogenesis of OSF and 10 of adjacent non-cancerous OSF tissue. Results:The expression of hTERT was negative in normal oral muscosal epithelium.The frequency of hTERT positive expression was higher in carcinogenesis lesions of OSF(12/14) than those in OSF(7/46)(P

Purpose: To observe the therapeutic effect of plum-blossom-magnet-needle synthetic therapy on benign prostatic hyperplasia (BPH). Method: 219 cases of BPH patients were randomly divided into two groups. There were 149 cases in treatment group, which treated by pressing, needling with plum-blossom-magnet-needle and putting effect-increasing pads on Prostate Reflex Area, bilateral Zhongliao (BL 32), Neck 1, Neck 7, Mingmen (GV 4) and Yongquan (KI 1). There were 70 cases in control group, which treated by orally taken Terazosin Hydrochloride (Gaoteling) tablets, 2 mg/time, twice a day. The therapeutic course was 4 weeks.After one therapeutic course, the therapeutic effect was evaluated. Results: The marked effect rate was 63.3% and total effective rate was 91.8%; in the control group, there were no markedly effective cases and the total effective rate was 72.7%. There was a significant difference between two groups (P<0.05). Conclusion: The plum-blossom-magnet-needle synthetic therapy had a significant therapeutic effect for the treatment of benign prostatic hyperplasia (BPH), so it was a safe and effective therapy for BPH.

Objective To observe the influence of peripheral serum came from patients with hepatectomy at different time points on hepatocyte proliferation in vitro. Methods According to the different types of cultured serum, cultured HL-7702 cells were divided into fetal bovine serum (FBS) group, preoperative serum group, 0.5 h, 3 h, 24 h and 72 h post operative serum groups. All groups of cells were cultured for 72 hours in the Cell-IQ unmarked living cell image analysis system, and the amplification curves of each group were mapped by continuous counting of cells. The cell amplification multiple was compared between all groups after culturing for 72 hours. BrdU immunofluorescence staining was performed and BrdU positive rate was calculated for comparing the cell proliferation of all groups. Results Amplification curves showed that HL-7702 cell proliferation rates of all human serum groups except for 72 h post operative group were higher than those of FBS group. Human serum 0.5 h and 3 h postoperative groups were more obvious. The amplification multiples of human serum groups, except for 72 h post operative group were all significantly higher than those of FBS group (P<0.01), and 0.5 h and 3 h post operative groups were both significantly higher than those of preoperative group (P < 0.05). BrdU positive rates of all human serum groups were significantly higher than those of FBS group (P < 0.01), which were significantly higher in 0.5 h and 3 h post operative groups than those of preoperative group (P < 0.05), but there were no statistical differences between 24 h and 72 h post operative groups and the preoperative group. Conclusion Human serum can promote the proliferation of hepatocytes compared with that of FBS. The influence of serum acquired post hepatectomy is closely associated with the post operative time.

Objective To analyze the imaging features and follow-up changes of high-resolution CT(HRCT) in Goodpasture syndrome.Methods HRCT imaging features and follow-up findings of 15 cases Goodpasture syndrome confirmed by clinical were analyzed retrospectrively.The imaging features included extent,forms and follow-up changes.Results The lung lesion of Goodpasture syndrome involved two lobes(n=1), three lobes(n=2), four lobes(n=5) and five lobes (n=7).Upper lobe of the right lung was the most common involved region.Centered on the hilum of lung consolidations confused ground glass opacity (GGO) were showed in 7 cases, GGO distribution of pulmonary leaflets in 5 cases.On follow-up observation, lobar or segmental consolidation could change into GGO,GGO could disappear in short times.Conclusion Multiple lobar or segmental consolidations confused GGO without the lung bottom and periphery involvement is the imaging characteristics of Goodpasture syndrome patients with anemia and hemoptysis.HRCT is a helpful method for the diagnosis and following up of Goodpasture syndrome.

Objective Temporal lobe epilepsy (TLE),frequently accompanied by working memory dysfunction,is an important form of adult epilepsy syndrome.Study on neural mechanisms of TLE with working memory dysfunction has important scientific value and clinical significance.Theta oscillations are synchronous activity of the brain in the 4-8 Hz frequency range.It is well recognized that there is a close relationship between the theta oscillations and working memory behavior.TLE patients with working memory dysfunction were taken as the research subjects,and the theta oscillations absence in TLE with working memory behavioral disorder was carried out,which provide the reference for further research into the neural mechanisms of TLE with working memory dysfunction.Methods The 34-channel electroencephalographs (EEGs) were recorded from TLE group (18 TLE patients) and control groups (18 healthy subjects) while subjects were performing visual working memory (delayed matching-tosample) tasks.The EEGs during the 3 s delay period was analyzed as experimental data.Fourier transform was used to assess the EEGs spectrum.The channel with the strongest spectrum was selected as the feature channel.Short-time Fourier transform algosiths was employed to calculate the time-frequency representation of the feature channel for the TLE and control groups.Frequency band with the strongest spectrum was selected for control group as the feature frequency band in working memory.Then,the topographical maps of the feature frequency band spectrum were calculated for the TLE and control groups.Results Compared with the control group,the working memnory behavioral performance of the TLE group was lower:accuracy of the TLE group was significantly lower than that of the control group (P＜0.05),and reaction time of the TLE group was significantly longer than that of the control group (P＜0.001).The feature channel in working memnory was frontal midline (Fz) and the feature frequency band was theta band.The Fz spectrum of the feature frequency for TLE was lower than that for control (P＜0.05).The frontal spectrum (seven channels) of the feature frequency for TLE was lower than that of control (P＜0.01).Conclusions Theta oscillations for TLE with working memory behavioral disorder is absent,which maybe one of the possible neural mechanism of TLE with working memory dysfunction.

The relationship between the precision of the estimated sampling variances and the number of samples was quantitatively established. It revealed that the product of the standard deviation of the estimated samplingvariance and square root of the number of samples was a constant. Monte Carlo simulation technique was employed to verify the equation. Populations with Gaussian,unifohn random and multi-nomial(taking silicon carbide material as an example) distributions were studied with satisfactory results.