Objective To establish a method for detecting the EGFR gene mutations by the real-time fluorescence quantification PCR combined with Sanger sequencing and to preliminarily explore its clinical application value .Methods With EGFR gene hotspot mutations region exon 19 and 21 as the research locus ,the specific amplification and the sequencing primer were designed ,the known wild-type and mutant samples were utilized to construct the corresponding plasmid as the standard substance by the TA clone technique .Then the EGFR gene mutation detection method by the real-time fluorescence quantification PCR combined with Sanger sequencing was established and the methodological and the application evaluation were performed .Results The wild-type and mutant standard plasmids of the EGFR gene exon 19 and 21 were constructed successfully .The EGFR gene mutations detection method of the real-time fluorescence quantification PCR combined with Sanger sequencing was established ,which had high sensitivi-ty(101copies/μL)andgoodrepeatability(intra-assayCVandinter-assayCVofthereal-timefluorescencequantificationPCRofex-on 19 and 21 were 1 .42% /3 .52% and 0 .97% /2 .44% ,respectively ) .20 clinical samples were simultaneously detected by this method and the traditional Sanger sequencing ,the results were completely consistent .Conclusion The EGFR gene mutations detec-tion method of the real-time fluorescence quantification PCR combined with Sanger sequencing is successfully established ,which can be used in the clinical sample detection .

Objective To evaluate the effect of subfascial endoscopic perforator surgery(SEPS) for the(treatment) of recurrent venous ulcer.Methods SEPS Was preformed on 56 limbs with recurrent venous(ulcer),and ulcer healing was assessed at follow up.Results Fifty-five limb ulcers healed in 10～49 days,one limb ulcer was not healed after 90 days,but healed after ligation of lesser saphenous vein.The patients were followed up for 1 to 3 years,1 case recurred after 6 months and it healed with antibiotic treatment;2 cases recurred after 18 months,and both healed after high ligation of lesser saphenous vein.Conclusions SEPS is effective and safe in the treatment of recurrent venous ulcer.SEPS is minitraumatic with few(complications).

Objective:To detect tenascin(TN) expressions and its effect in salivary gland tumor. Methods:The expressions of TN was examined in 8 chronic inflammations of salivary glands, 50 benign adenomas and 57 malign adenomas by Envision immunohistochemical technique. Results:Expression level of TN in the malign adenomas was higher than that in benign adenoma and chronic sialadenitis; higher in low differentiation than that in high differentiation and higher in lymphatic metastasis groups than that without lymphatic metastasis groups. No obviously difference was found between soakage inside the blood vessel and nerves and without soakage. Conclusion: TN might regulate cell attachment of epithelial cells in adenomas, and involve in adenomas' histogenesis and development.

Health Knowledge, Attitudes, Practice ; Humans ; Occupational Diseases ; prevention & control ; Rural Population ; Sampling Studies ; Surveys and Questionnaires ; Transients and Migrants

Denys-Drash Syndrome ; diagnosis ; Humans ; Infant ; Male

OBJECTIVETo investigate protective effect of paeonol (Pae) on human umbilical vein endothelial cells (HUVECs)injured by hyperlipidemic serum and explore its possible mechanism.METHODS The injury model was induced by 20％ hyperlipidemic serum incubating HUVECs for 24 h.Pae 124,247 and 495.μmol· L-1 were given followed by administration of hyperlipidemic serum for 24 h.The morphological changes were observed under inverted microscope,cell survival rate was evaluated by MTT method,the nitric oxide (NO) content was measured by nitric acid reductase method and the endothelial nitric oxide synthase (eNOS) mRNA expression was determined by RT-PCR.RESULTSCompared with normal contorl group,most cells in model group split and exfoliated.However,the morphology was tending to normal level after intervention with Pae.Pae significantly improved cell viability(P ＜0.01).Compared with model group,the survival rate increased from (53.0 ±10.1 ) ％ to ( 68.4 ± 9.1 ) ％,( 84.5 ± 6.7 ) ％ and (98.1 ± 7.5 ) ％,respectively.The NO content and eNOS mRNA expression both increased greatly in Pae groups(P ＜ 0.01 ).Compared with model group,content of NO increased from 54 ± 4 to 79 ± 6,115 ± 5 and ( 136 ± 6) μmol · L- 1,respectively.The expression level of eNOSmRNA improved from 0.215 ± 0.060 to 0.451 ± 0.045,0.563 ± 0.013,0.704 ± 0.068,respectively.CONCLUSIONPae exerts protective effect on HUVECs injured by hyperlipidemic serum by increasing eNOS mRNA expression,which might therefore improve the content of NO.

Objective To evaluate results of double eyelid operations with different incisions.Methods 100 patientswere selected randomly from out-patients that accepted double eyelid operation.Half of them were objected to double eyelid operation through a palpebral marginal incision.Incision was designed along the palpebral margin,close to boots of eyelashes.Most of the subcutaneous tissues was cut and removed,and then dermis and tarsal plate were fixed with 5 stitches of suture.About 1-3 mm wide of skin along superior margin was removed reasonably.In another 50 cases.double eyelid operation was performed with conventional incision.Incision was designed 6-8 mm far from the palpebral margin.Most of the subcutaneous tissues was removed.and then dermis and tarsal plate were fixed with 5 stitches of suture.Results Postoperative results were evaluated 5 days and 1 month after operation respectively.Postoperative double eyelid shapes were all satisfled in two groups.Swollen eyedids and scar were more inconspicuous in the cases with the palpebral-marginal incision without impairment on eyelashes.Conclusion Compared with double eyelid operation with conventional incision,blepharoplasty with palpebral margin incision has the same results but with lighter impairment,milder hydropsia and more inconspicuous scar.The key point of this procedure is the fixation of dermis and tarsal plate,keeping a good radian of double eyelid line.

Objective To explore the causal influences between activated regions in executive control network when participants were performing attention network test (ANT)under passive hyperthermia using Granger causality analysis.Methods Thirty male healthy participants which were randomly divided into control group (n=1 5)and experimental group (n=1 5)performed ANT task during two thermal conditions:hyperthermic condition (50℃)and normothermic condition (20℃)during functional magnetic reso-nance imaging.The data processing was performed using statistical parametric mapping 8 (SPM8).Granger causality analysis was performed on the resulted activated regions,as well as the correlations of the GC values with the reaction time in both groups.Results Between-group comparisons on Granger causality analysis revealed significantly enhanced influences of anterior cingulate gyrus and right inferior parietal lobule on dorsolateral prefrontal cortex,and bidirectional influences between dorsolateral prefrontal cortex and right superior parietal lobule,but decreased influence between right superior parietal lobule and right inferior parietal lobule.The correlation analysis showed significant correlation between GC value of right superior parietal lobule to dorsolateral prefrontal cortex and the reaction time in control group (r=-0.558,P =0.031),but yet there was no significant correlation in experimental group (r=0.355,P =0.257).Conclusion Heat stress affected the causal influences within the executive control network,which subse-quently disrupted the executive control performance during passive hyperthermia.

Objective To observe effects of aqueous extractive of Clarke boea herb on mice with gastric ulcer, and provide basis for clinical application.Methods Gastric ulcer model was induced by reserpine. The mice were divided into normal group, model group, ranitidine group, low and high dose of Clarke boea herb groups. Mice in treatment groups were given corresponding drugs by gavage, and the normal group and control group were given the same amount of saline for 4 weeks. The area of gastric ulcer, serum proinflammatory cytokines gastric juice volume, gastric juice acidity, iNOS and COX-2 gene expressions were detected.Results Compared with control group, the gastric ulcer area in low and high dose of Clarke boea herb groups decreased significantly. Clarke boea herb increased the SS and VIP, decreased the MOT and SP serum levels (P<0.05,P<0.01). Clarke boea herb reduced the levels of serum proinflammatory cytokines of IL-6, IL-12, TNF-α and IFN-γ(P<0.05,P<0.01). Clarke boea herb also decreased the genetic expressions of iNOS and COX-2 (P<0.05,P<0.01).ConclusionClarke boea herb has good gastric ulcer preventive effects, and the high dose of Clarke boea herb group showed the best efficiency.

Objective To explore the effects of erythropoietin (EPO) on the expression of signal transducer and activator of transcription (STAT) 1,phosphorylated STAT1 (P-STAT1),STAT3,P-STAT3 and cell apoptosis in rat models of focal cerebral ischemia-reperfusion.Methods Eighty male SpragueDawley rats were randomly and evenly divided into four groups by completely random design method: shamoperation (group A),cerebral ischemia-reperfusion (group B),cerebral ischemia-reperfusion ± saline (group C) and cerebral ischemia-reperfusion ± EPO (group D).The model of focal cerebral ischemiareperfusion injury was established by blocking the left middle cerebral artery.All rats underwent MRI for the detection of the changes of infarct area between 2 h post ischemia and 24 h of reperfusion.Western blot was used to observe the expression of STAT1,P-STAT1,STAT3,P-STAT3.Terminal oxynucleotidyl transferase mediated dUTP biotin nick end labeling (TUNEL) was used to evaluate the cell apoptosis including the relative area (ROI area/whole brain area of the same layer × 100％) of abnormal signal region,relative optical density (rOD) and apoptotic index.One-way analysis of variance and q test were used to analyze the data.Results On T2WI imaging,rats in group B and group C presented large hyperintense areas in the cortex and subcortex of left hemispheric ((28.00±4.60)％ and (29.70±4.80)％ respectively).Group D presented less hyperintense areas in the cortex and subcortex of left hemispheric compared with group B and group C ((21.10±2.40) ％; F=11.285,P＜0.01).The expression of STAT1 and STAT3 proteins was not significantly affected by ischemia-reperfusion and EPO intervention compared with normal brain tissue (F=0.806,1.558,both P＞0.05).However,the level of P-STAT1 was low in group A (rOD =0.75±0.13) but increased after cerebral ischemia-reperfusion.Compared with group B and group C,P-STAT1 expression was lower in group D (B-D: 2.08±0.15,2.05±0.16,1.92±0.05; F=3.274,P＞0.05).The level of P-STAT3was also low in group A (rOD=1.02±0.09).Compared with group B and group C,P-STAT3 expression in group D was significandy higher (B-D: 2.22±0.13,2.04±0.14,4.21±0.21 ; F=40.719,P＜0.01).The apoptotic index of group B and group C was (42.00±1.30)％ and (41.20±2.50)％,respectively,which was significantly higher than that of group D ((20.90± 1.46) ％ ; F=378.704,P＜0.01).Conclusion Intraperitoneal injection of EPO can reduce the cerebral ischemic area and the number of apoptotic cells in the ischemic penumbra in rat ischemia-reperfusion models through increasing P-STAT3 and decreasing P-STAT1 levels.