Objective To express and purify the killer immunoglobulin-like receptor KIR3DL1 extracelluar domain.Methods pUC57-KIR3DL1 was used as template,KIR3DL1 extracelluar domain was amplified by polymerase chain reaction(PCR)and cloned into pGEM-T vector with A-T cloning technique.After DNA sequence analysis,the target fragment was inserted into the prokaryotic expression vector pET28a-DsbA to construct the recombinant vector pET28a-DsbA /KIR3DL1.The recombinant plasmid was transformed into E.coli BL21(DE3),and induced with IPTG.Bacterial pellets were resuspended in 8M urea and centrifuged to remove the insoluble material.The crude extract was purified by passing over a Ni-NTA-agarose column.After the inclusion body flowing through the Ni-NTA-agarose affinity chromatography was refolded successfully,it was purified by Superdex75 gel filtration and the purification effects of the fusion protein were identified by SDS-PAGE and western blot.Result Some fusion proteins were expressed in the supernatant,and the others were expressed in the form of inclusion bodies.The purity of fusion protein was over 95% after purification under denaturing condition.Conclusion The highly efficient expression of KIR3DL1 extracelluar domain laid the foundation for the further studies on exploration of the mechanism of immunization recognition between KIR3DL1 and its ligand.

Objective To study the current research status of minichromosome maintenance protein 2 (MCM2), the cell cycle proliferation marker, in the diagnosis of colorectal carcinoma. Methods Literatures about the application of MCM2 in the study of colorectal carcinoma were collected and reviewed.Results MCM2, as a marker of cell dysplasia and malignancy, was usually used in the study of carcinoma. The study on expression of MCM2 in the cell of colorectum in different proliferational stage might help to screen colorectal carcinoma as early as possible. Conclusion As a relatively specific and sensitive marker of cell proliferation, MCM2 might become a promising mark for diagnosing colorectal carcinoma in the early stage.

?AIM: To compare the clinical effect of 23G and 25G+vitrectomy for retinal detachment.?METHODS:Forty seven patients with retinal detachment were treated with 23G vitrectomy (27 eyes in 27 cases as group A) and 25G+ vitrectomy (20 eyes in 20 cases as group B ) . The operation time and the incidence of intraoperative complications were recorded. The occurrence of retinal reposition, visual acuity, intraocular pressure ( IOP ) and complications were observed. Postoperative follow-up time of the two groups were 3d, 1wk, 3mo. The relevant records were statistically analyzed and compared.?RESULTS: The operation time of 23G group and 25G+group were 50. 21+4. 52min, 49. 15+5. 14min,respectively and there was no significant difference between the two groups (P>0. 05). The main complications were retinal hemorrhage and iatrogenic retinal hole. There were 3 eyes with retinal hemorrhage, 2 eyes with iatrogenic retinal hole in 23G group, and 1 eye with retinal hemorrhage, 1 eye with iatrogenic retinal hole in the 25G+group, and the difference was statistically significant ( P<0. 05). The postoperative visual acuity of 23G group and 25G + group were significantly improved, and the differences between the two groups were not statistically significant at different time points after operation ( P>0. 05). The number of eyes with hypotonia in 23G and 25G+group were 3 and 1 eyes respectively, the difference was statistically significant ( P < 0. 05 ). But there were no significant differences between the two groups on IOP at 1wk and 3mo after surgery (P>0. 05). At the last follow-up, the results showed that 26 eyes ( 96%) with retinal reposition in 23G group, 19 eyes (95%) in 25G+ group, the difference was not statistically significant (P>0. 05).?CONCLUSION: The clinical effect of 23G and 25G+vitrectomy for retinal detachment is similar, but 25G+vitrectomy can reduce incidence of complications and early postoperative low IOP.

Objective To analyze laboratory turnaround time (TAT) and find effective ways to shorten TAT.Methods Data associated with cardiac panel (CK,cTnI and Mb) were collected in 2011 including 19 906 outpatient data and 22 973 inpatient data.The medians and the average medians of the quality indicators on TAT were calculated and the results were transformed to the Six Sigma scale to estimate the degree of control over related process.Processes were considered well controlled when σ ≥4.Based on the results of data analysis,an improvement plan was decided by laboratory quality management meeting and clinical communication meeting.The effect of the improvement plan was evaluated through 2011-2012 satisfaction surveys of outpatients and clinicians.Results The average median of overtime reports for outpatient from specimen collection to reception was 2.78％ (3.5σ),and 17.82％ (2.5σ) for inpatients.The average median of overtime reports for outpatient from specimen reception to result reporting was 3.39％ (3.4σ),and 2.96％ (3.4σ) for inpatients.The average median of overtime reports for outpatient from specimen collection to result reporting was 3.93％ (3.3σ),and 12.18％ (2.7σ) for inpatient.The results of TAT satisfaction surveys for outpatients from 2011 to 2012 were similar,which were 78％ in 2011 and 79％ in 2012; the results for clinicians showed an increase from 80％ in 2011 to 90％ in 2012,including an increase from 75％ to 79％ for very satisfaction choice.Conclusions Outside the laboratory TAT is a key step in sickroom patients delay TAT.The implementation for ten improvement suggestions enabled to shorten TAT effectively.

Objective To investigate the mechanisms of phenytoin accelerating healing process.Methods Adult male Wistar rats that survived ligatation of the left coronary artery were randomized to phenytoin group or operation control and compared to sham-operated rats.The time-dependent proteolytic activity of MMP-2 and MMP-9 were detected by gelatin zymography.Picrosirius red staining plus polarized light micrscopy was used for qualitative and quantitative analysis of collagen include collagen volume fraction(CVF) and ratio of typeⅠ/Ⅲ collagen.In addition,infarct thickness and myocyte cross-sectional area were also evaluated by image analysis.Results Fourteen days after myocardial infartion(MI),phenytoin reduced infarct wall thinning.Compared with control group,the rats received phenytoin had less myocyte cross-sectional area.Two weeks after MI,CVF in two groups both had significantly dynamic increase and phenytoin accelerated the change.In contrast to control group,ratio of typeⅠto type Ⅲ collagen in phenytoin increased more quickly.Apart from these results,phenytoin did little to CVF in non-infarcted region.Analysis of MMPs activity in myocardial extracts by zymography demonstrated that infarction-induced expression of proMMPs and active MMPs was both upregulated in phenytoin group and operation control.We found that after MI,MMP-9 activity increased as early as 1 day and reached a maximum then gradually descended,whereas MMP-2 began to increase rapidly and remain elevated for up to 14 days thereafter.Phenytoin seemed toenhance expression of MMP-2 and MMP-9.1 day after MI,active MMP-9 in phenytoin group expressed an increasing trendency compared to MI control.Conclusion Phenytoin can attenuate the degree of post-infarction left ventricular dilation and expansion of the infarct during the early phase of MI healing.MMP-2 and MMP-9 enhanced by phenytoin probably played a prominent role.

Objective To investigate the expression pattern of Nrf2 in the lung of septic rat and preliminary analysis of the role of Nrf2 in the development of sepsis. Methods Wistar rats were used in this study, it was divided into 4 groups, including normal control group, pure burn group, burn with staphylococcus sepsis group, burn with pseudomonas sepsis group. According to the different time intervals such as 2 hours, 8 hours, and 24 hours, it was divided into three sub-group after injection of bacteria. The expression of Nrf2 in the lung at different time intervals was determined. Results Nrf2 mRNA in the lungs of normal rats was high expression （74.0±7.0）, Nrf2 mRNA in the lungs of pure burns rats obviously down-regulated, respectively as 34.5±1.9,50.4±2.2,32.1±1.4, （t＝5.69～14.63,P＜0.01）. Nrf2 mRNA in burn sepsis caused by Staphylococcus aureus in lung tissue of rats down-regulated expression, respectively as 53.1±5.0,14.4±1.6,48.5±1.9,and reached peak at 8 h（t＝5.59～29.3,P＜0.01）. Pseudomonas aeruginosa burn sepsis didn't induced Nrf2 mRNA in the lung tissue, but it showed a downward trend at 2h（71.0±8.1,P＞0.05）and markedly reduced after 8, 24 h（24.8±2.1,4.1±2.0,t＝21.33,68.1,P＜0.01）.Conclusions The distribution of Nrf2 mainly localized in immune organ, and it directly took part in the post burn immune response.

BACKGROUND: Smad7 is a major repressible protein in transforming growth factor β(TGF-β1) signal transduction pathway,which possess antifibrotic effects.OBJECTIVE: To construct rat Smad7 eukaryotic vector and to observe the mRNA expression level of TGF-β1, collagen Ⅰ and collagen Ⅲ in rat hepatic stellate cells (HSC)-T6 cell.DESIGN, TIME AND SETTING: The gene recombination and cytology observation experiment was performed at the First Affiliated Hospital of Shihezi University School of Medicine.MATERIALS: pcDNA3.1 (+) plasmid was reserved in the laboratory. E coil DH5α was presented by Key Laboratory of Xinjiang Endemic and Ethnic Diseases, Shihezi University School of Medicine. The HSC T6 cell was provided by Cancer Institute and Hospital, Chinese Academy of Medical Sciences.METHODS: Rat Smad7 cDNA was cloned into eukaryotic plasmid pcDNA3.1(+) to construct Smad7/pcDNA3.1(+) plasmid and transfected it into HSC-T6 ceils by Lipofectmine 2000. The experiment was divided into normal control, empty vector and Smad7 transfected groups, and the positive cells were selected by G418.MAIN OUTCOME MEASURES: The levels of Smad7, TGF-β1, collagen Ⅰ and Ⅲ mRNA was detected by reverse transcriptase polymerase chain reaction, respectively.RESULTS: Smad7 eukaryotic vector was successfully constructed and confirmed by endonuilease digestion and sequencing. Compared to the control and empty vector groups, Smad7 mRNA expression was significantly higher in Smad7 transfected group (P < 0.01 ); and TGF-β1 and collagen Ⅰ mRNA expression was notably reduced (P < 0.01). There was no statistically significant difference of the change of collagen Ⅲ mRNA expression among the three groups (P>0.05). The difference of Smad7, TGF-β1,collagen Ⅰ and Ⅲ mRNA expression had no statistically significant between control and empty vector groups (P_(all) > 0.05)CONCLUSION: Smad7 eukaryotic expression vector is successfully constructed. The Smad7 gene can effectively expressed in transfected HSC-T6 cell, and decrease mRNA expressions of TGF-β1 and collagen Ⅰ.

Objective To investigate the relationship of tumor malignancy with Na+ microcurrent measured by microcurrent detector in order to develop a novel approach for rapid intraoperative diagnosis of tumor.Methods 101 cases of various tumors were studied.Na+ microcurrent of tumor tissue or surrounding normal tissue was measured by Na+ microcurrent detector.All tumor tissues were sampled for histopathological examination as well.Results In 97 cases(96%),the Na+ microcurrent level in tumor tissue was significantly different from that in surrounding normal tissue.It was greater than 30?A in tumor tissue with the maximum of 60-70?A,and averagely 20?A in surrounding normal tissue.The consistency of Na+ microcurrent detection and pathological examination was 96%.Conclusion the measurement of Na+ microcurrent by microcurrent detector is rapid,convenient and accurate,which may become a novel approach for rapid intraoperative diagnosis of tumor malignancy.

Objective To investigate the gender difference in the smile aesthetic features in aged Han Chinese with normal occlusion and its clinical significance.Methods Fifty-six aged Han Chinese (male:28,female:28,aged 60-66 years)with normal occlusion were included in this study.Smiling images of the resting-position and mandibular postural position were obtained in an anteriorposterior view.The CoSmileMAA1.0 software was used to evaluate the indices associated with the smile-aesthetic features,and its clinical significance was analyzed.Results Significant differences were noted in the nasal ala width,upper lip length,lip clearance,angle oris width in resting-position/ smiling position together with the changes of lip clearance,angle oris width,distance of inferior margin of upper lip to cutting edge of the maxillary incisor teeth,distance of superior border of lower lip to cutting edge of the maxillary incisor teeth,smile line ratio and type of smile (all P＜0.05).When the subjects were smiling,the changes of lip clearance was greater in females than in males [(10.7±1.9)mm vs.(11.3±1.6)mm,P＜0.05],the changes of angle oris width was greater in males than in females [(14.1±1.6)mm vs.(13.4±1.3)mm,P＜0.05],and the smile line ratio was less in males than in females [(0.9±0.2) vs.(1.1±0.5),P＜0.05],which indicate that the females had more attractive smile in aged Han Chinese with normal occlusion than males.The type of smile was mainly high smile in elderly females and median smile in elderly males [53.6％ (15 cases) and 60.7％ (17 cases),x2 =6.43,P＜0.05].Conclusions Significant gender difference is noted in the smile-aesthetic features in aged Han Chinese with normal occlusion.Modulation of maxillary incisor teeth length and smile line ratio can contribute to the aesthetic appearance of smiling.

Objective: Our aim was to investigate the alteration of p16 gene in human pancreatic carcinoma. Methods: A total of 66 human pancreatic tissue specimens, comprising 51 with pancreatic carcinomas and 15 normal pancreatic tissue specimens, were examined for homozygous deletion and mutation of p16 gene by using PCR-SSCP method. Results: No mutation and deletion was detected in 15 normal pancreatic tissue samples. Of 51 pancreatic carcinoma specimens, only one was found mutation for p16 gene in PCR-SSCP assay, and the deletion of the p16 gene in 23 samples were confirmed by using PCR, with a 45% p16 gene deletion rate. Conclusion: These data suggest that p16 gene alterations may play a role in the progression of human pancreatic carcinoma.