Objective:To investigate the expression of Gli-2 protein and nuclear proliferation marker Ki-67 in human thymic tu-mors, as well as its relationship with clinicopathological features and prognosis. Methods: Immunohistochemical staining was per-formed to determine the expressions of Gli-2 and Ki-67 in 64 thymic tumor cases, in which 9 were type A, 6 were type AB, 11 were type B1, 22 were type B2, and 16 were type C. Results:1) Positive expression rates of Gli-2 in types A, AB, B1, B2, and C thymomas were 1/9 (11.11%), 2/6 (33.33%), 2/11 (18.18%), 5/22 (22.73%), and 13/16 (81.25%), respectively. Statistically significant differences existed in the two sets of data (P<0.05). Positive rates of Gli-2 in the Masaoka stages Ⅰ, Ⅱ, Ⅲ, and Ⅳ were 2/10 (20%), 12/41 (29.27%), 4/6 (66.67%), and 5/7 (71.43%), respectively. The statistical differences of these two sets of data were also significant (P<0.05). 2) The expression levels of Gli-2 in tumor tissues were closely correlated with the pleura (P<0.05) but not associated with gen-der, age, myasthenia gravis, and necrosis with hemorrhage (P>0.05). 3) The positive labeling indexes of Ki-67 in invasive and non-inva-sive thymomas were 7.14 ± 6.99 and 15.24 ± 9.13, respectively. The differences between both thymomas were statistically significant (P<0.05). 4) A positive correlation existed in the expression of Ki-67 and Gli-2 in thymomas. Five-year progression-free survival (PFS) rate was lower in the Gli-2 positive group (56.5%, 13/23) than in the negative group (92.7%, 38/41). The Ki-67-positive group (61.5%, 16/26) also showed a lower five-year PFS than that in the negative group (92.1%, 35/38), with statistically significant differences be-tween the two groups (P<0.05). Multivariate Cox's proportional hazard regression analysis indicated that the Gli-2-positive group ex-pression, Ki-67-positive group expression, and invasion of the pleura were independent prognostic factors of thymic tumors. Conclu-sion:The expression of Gli-2 and Ki-67 showed a positive correlation with thymic tumors. Detecting the combined expression of Gli-2 and Ki-67 may elucidate the clinicopathological features and prognoses of patients with thymic tumors.

Objective To analyze the diagnosis and prognosis value of abdominal CT scans in patients with acute pancreatitis complications.Methods 151 cases with acute pancreatitis were selected.The relationship between abdominal CT performances and the common complications and death in patients was analyzed.The prognosis of patients with different Balthazar CT severity index (CTSI) grade was compared.Results The complication rate of patients with CT signs of fatty liver, pleural effusion, liver gap effusion, adrenal gland involvement (AGI), penirenal space involvement (PSI) and gastric bare area involvement (GBAI) was significantly higher than that of patients with negative CT findings above (P<0.05).With CTSI grading increasing, the patient''s fasting time, heating time, hospital stay, recovery time of blood amylase were extended, and the incidence of pseudo cyst, transit surgery, organ failure and death was gradually increasing (P<0.05).Conclusion Severe fatty liver, AGI, GBAI, PSI and liver gap effusion are risk factors for acute complications and death in patients with pancreatitis.

Objective To observe the clinical efficacy of dye laser combined with Xin'an repair solution in the treating superficial infantile hemangioma(IH). Methods 100 patients with superficial IH were randomly divided into observation group and control group.The observation group received dye laser combined with Xin'an re-pair solution,while the control group was treated only with dye laser.The clinical efficacy and the rate of complica-tions after treatment were compared between the two groups,and the different efficacy between the head neck group and the non head neck group in the observation group was further analyzed. Results The total effective rate of the observation group(90%)was significantly higher than the control group(70%),and the difference was sta-tistically significant(P<0.05).The rate of complications in the observation group was significantly lower than that in the control group(6%vs.32%,P<0.05).In the observation group,the total effective rate of head neck group was better than that the non head neck group(P<0.05). Conclusion Dye laser combined with Xin'an repair so-lution is effective in the treatment of superficial IH,and it can significantly reduce complications.It is evidently ef-fective for head and neck superficial IH.

Objective To investigate the clinicopathologic characteristics and relevance of main and minor lesions of synchronous multiple early gastric cancers ( SMEGC) and gastric high grade intraepithelial neoplasia ( GHGIN) . Methods Thirty-two patients with SMEGC or/and GHGIN who were diagnosed and treated with endoscopic submucosal dissection in Nanjing Drum Tower Hospital from July 2012 to September 2016 were enrolled in this study. Their clinicopathologic characteristics were summed up, and the correlation between main and minor lesions on the size, location, endoscopic classification, pathologic type, invasion depth and vascular invasion were analyzed. Results Among the 32 patients, with mean age of 66. 19±7. 46 years, 90. 62％(29/32) were male, 17 cases (53. 3％) had family history of gastric cancer, 25 (78. 13％) had smoking history, and 22 ( 68. 75％) were alcohol users. There were 30 cases ( 93. 75％) and 31 cases ( 96. 88％) with mucosal atrophy and intestinal metaplasia, respectively. The size of main and minor lesions showed a positive correlation (r=0. 4167, P=0. 018). The endoscopic classification of major and minor lesions had no statistical significant consistency ( P=0. 314 ) , but the pathologic type and invasion depth between major and minor lesions demonstrated a moderate significant positive correlation ( P<0. 05 ) . The comparison of location between the main and minor lesions did not show correlation. However, it showed a significant correlation between major lesion which on the upper 1/3 of stomach and minor lesion on the lower 1/3 of stomach ( r=0. 463,P=0. 003) . Further more, when the main lesion was at posterior gastric wall, the minor lesions on lesser curvature were increased, which showed a positive correlation( r=0. 417,P=0. 009) . Conclusion Old-age male with long-term smoking and alcohol history whose lesions combined with surrounding mucosa merger atrophy and intestinal metaplasia are considered as a high risk group in patients with SMEGC or/and GHGIN. Therefore, clinicians must keep high vigilant and make carefully observations on this group of patients during endoscopic examination, and consider the correlation between main and minor lesions to avoid misdiagnosis.

Cell-free transcription and translation (TXTL) system is a cell extract-based system for rapid in vitro protein expression. The system bypasses routine laboratory processes such as bacterial transformation, clonal screening and cell lysis, which allows more precise and convenient control of reaction substrates, reduces the impact of bacteria on protein production, and provides a high degree of versatility and flexibility. In recent years, TXTL has been widely used as an emerging platform in clusterd regularly interspaced short palindromic repeat (CRISPR) technologies, enabling more rapid and convenient characterization of CRISPR/Cas systems, including screening highly specific gRNAs as well as anti-CRISPR proteins. Furthermore, TXTL-based CRISPR biosensors combined with biological materials and gene circuits are able to detect pathogens through validation of related antibiotics and nucleic acid-based markers, respectively. The reagents can be freeze-dried to improve portability and achieve point-of-care testing with high sensitivity. In addition, combinations of the sensor with programmable circuit elements and other technologies provide a non-biological alternative to whole-cell biosensors, which can improve biosafety and accelerate its application for approval. Here, this review discusses the TXTL-based characterization of CRISPR and their applications in biosensors, to facilitate the development of TXTL-based CRISPR/Cas systems in biosensors.
CRISPR-Cas Systems ; Bacteria

Objective To investigate biomechanical differences of two posterior occipitocervical internal fixation techniques for treating basilar invagination with atlantoaxial dislocation (BI-AAD). Methods Intra-articular cage + posterior occipital plate+C2 pedicle screw (Cage+C2PS+OP), and intra-articular cage+C1 lateral mass screw+C2PS (Cage+C1LMS+C2PS) models were established based on occipitocervical CT data of the BI-AAD and clinical operation scheme, and the stability of atlantoaxial joint and stress distribution characteristics of C2 endplate and implanted instruments under different motion states were analyzed. Results Compared with the Cage+C1LMS+C2PS model, the atlantoaxial range of motion ( ROM) under flexion, extension, lateral bending and axial rotation in the Cage+C2PS+OP model were reduced by 5. 26% , 33. 33% , 43. 75% , -5. 56% , and stress peak of screw-rod fixation system were reduced by 47. 81% , 60. 90% , 48. 45% , 39. 14% , respectively. Under two internal fixation modes, stresses of C2 endplate and cage were mainly distributed on the compressive side during the motion, and both the screw-bone interface and the caudal side of screw subjected to large loading. Conclusions Two internal fixation methods could provide similar stability. However, the stress concentration of screw-rod system was more obvious and the possibility of screw loosening and fracture was greater under Cage+ C1LMS+C2PS fixation.

Objective: To explore the effects of ApoE epsilon4 (ApoE-ε4) alleles on cognitive function and resting-state functional MRI (rs-fMRI) in patients with amnestic mild cognitive impairment(aMCI) based on a prospective cohort study. Methods: An average of 20 months of prospective observations were conducted on 16 ApoE-ε4-carriers and 24 non-carriers of aMCI. Neuropsychological assessments and rs-fMRI data were collected at both baseline and follow-up. All participants were assessed by a battery of neuropsychological tests and underwent rs-fMRI. Two core regions of the default mode network (DMN), the left posterior cingulate cortex (PCC) and the medial prefrontal cortex (mPFC), were selected as seeds to calculate the functional connectivity. Two-way repeated measures analysis of variance was used to assess the effects of ApoE genotype(ε4-carriers, nonε4-carriers), interval and the interaction between these two factors for functional connectivity extracted from changed region found by t-test.Conversion rates of dementia were compared between ApoE-ε4-carriers and nonε4-carriers at follow-up using Chi-square test. For the comparison of functional connectivity and clinical data between ApoE-ε4-carriers and nonsε4-carriers in baseline and follow-up, the normal distribution test was carried out first. If the normal distribution was fitted, the two-sample t test was used, otherwise, the Mann-Whitney rank sum test was used. Finally, the general linear model was used to assess the relationships between alterations in functional connectivity and in neuropsychological assessments as well as the interaction effect. Results: (1)Significant decline in memory domains were found in ApoE-ε4-carriers as compared to non-carriers at both baseline and follow-up. The ApoE-ε4-carriers (14/16) presented a higher conversation rate than non-carriers(13/24, χ²=4.862, P=0.027) at follow-up. (2)Functional imaging analysis revealed that ApoE-ε4-carriers exhibited significantly higher functional connectivity between the left PCC and the left angular (ApoE-ε4-carriers: 0.23±0.11, non-carriers: -0.03±0.13, t=4.800, cluster size: 1 944 mm³, P=0.004), and between the left mPFC and the left angular (ApoE-ε4-carriers: 0.33±0.21, non-carriers: 0.08±0.18, t=5.040, cluster size:1 836 mm³, P=0.006) as compared to non-carriers at follow-up. We detected significant effect for the interaction interval by ApoE-ε4 on functional connectivity between the left angular and the left PCC (F=10.833, P=0.002)as well as the left mPFC (F=7.280, P=0.010). (3)The alteration of functional connectivity value between the left mPFC and the left angular in ApoE-ε4-carriers was positively correlated with the changes ofimmediate memory (r=0.692, P=0.018). The correlation was not statistically significant in ApoE-ε4-noncarriers (r=-0.198, P=0.417) and the integration effect was significant (F=8.632, P=0.006). Conclusions The ApoE-ε4 actually accelerates the deterioration of cognitive function in aMCI patients and carriers presented relatively reserved functional connectivity between the left angular and other core regions within DMN, which indicated the disruption of functional connectivity may be one of the underline mechanisms of ApoE-ε4 during AD process.