Objective To investigate the expressions of advanced glycation end products (AGEs) and their receptors in keloid. Methods Serum and skin tissue specimens were collected from 20 patients with keloid, 20 patients with hyperplastic scar and 20 normal human controls. Fluorospectrophotometer was used to measure the serum level of AGEs, and immunohistochemistry and Western blotting to detect the expressions of AGEs and AGER in skin tissue specimens. Results The serum level of AGEs was (0.713 ± 0.098) AU/ml and (0.699 ± 0.077) AU/ml respectively in patients with keloid and those with hypertrophic scar, significantly higher than that in normal controls (0.179 ± 0.056 AU/ml, F = 283.82, P < 0.01 ). A positive expression of AGEs and AGER was observed in tissue specimens of keloid and hyperplastic scar, but not in the control specimens. As Western blotting showed, the expressions of AGEs and AGER were higher in tissue specimens of keloid and hyperplastic scar than in the control specimens (F = 18.04, 42.80, both P < 0.05), while no significant difference between keloid and hyperplastic scar tissue specimens was observed (P> 0.05). Conclusion There is a high expression of AGEs and AGER in keloid, which may contribute to the development of keloid.

Objective To compare different noninvasive models in diagnosis of liver fibrosis in patients with chronic hepatitis B (CHB).Methods A total of 838 CHB patients admitted in Shenyang Sixth People’ s Hospital during March 2003 and October 2013 were enrolled in the study.All the patients received liver biopsy, blood and ultrasound examinations; the AST-to-ALT ratio ( AAR), AST to platelet ratio index ( APRI) , age platelet index ( API) , cirrhosis discriminant score ( CDS) , spleen to platelet ratio index ( SPRI) and age-spleen to platelet ratio index ( ASPRI) were obtained.Area under receiver operating characteristic curve (AUROC) was used to assess the clinical value of noninvasive models in diagnosis of significant liver fibrosis (S2-4), advanced liver fibrosis (S3-4) and early liver cirrhosis (S4).Results Among six noninvasive models, APRI had the lowest value of AUROCs ( <0.7), while ASPRI had the highest AUROCs value in diagnosis of liver fibrosis.The AUROCs of ASPRI in diagnosing significant liver fibrosis, advanced liver fibrosis and early liver cirrhosis were 0.861, 0.873 and 0.881 with the sensitivities of 69.4%, 76.9%and 87.0%, the specificities of 87.9%, 81.5% and 75.8%, the positive predictive values of 90.9%, 74.9%and 46.1%and the negative predictive values of 62.2%, 83.1%and 96.1%, respectively.Taking<5.2 and≥9.7 as the cut-off values for exclude significant liver fibrosis and diagnosis of significant liver fibrosis, respectively, 49.4%(414/838) of the patients may avoid liver biopsy with an accuracy of 92.3%(382/414).Conclusion ASPRI is of value in diagnosing significant liver fibrosis and early liver cirrhosis in patient with chronic hepatitis B, and the number of liver biopsy can be reduced.

Objective To investigate the expressions of advanced glycosylation end products (AGE) in skin of mice with diabetes mellitus (DM) for different durations,and to evaluate their influence on collagen fibers.Methods Forty healthy 8-week-old male C57BL/6J mice were divided into DM group (n =20) and control group (n =20) to receive multiple intraperitoneal injections of low dose streptozotocin (50 mg/kg) and citric acid buffer (0.1 mol/L),respectively,for 5 consecutive days.Ten mice were sacrificed in each group on week 4 and 12 respectively after the last intraperitoneal injection,and full-thickness skin tissue samples were harvested from the middorsal region of each mouse.Then,hematoxylin-eosin (HE) staining was performed to observe histological changes,and total collagen content was estimated according to hydroxyproline content measured by an alkalinehydrolysis method.The cross-linking degree of collagen was determined by Edman degradation method using pepsin,the mRNA expression level of collagen type Ⅰ and Ⅲ by real-time quantitative PCR,the content of AGE by fluorospectrophotometry and Western blotting,and the level of malondialdehyde (MDA) by using a thiobarbituric acid method.Statistical analysis was carried out by t test.Results As light microscopy showed,the skin became obviously thinner in the diabetic mice with a progressive decrease in the number of collagen fibers in comparison with the control mice.On week 4 and 12 after the last injection,the diabetic mice exhibited a significant reduction in the content of hydroxyproline ((684.5 ± 76.7) vs.(787.7 ± 87.7) rg/g,(558.1 ± 73.1) vs.(757.8 ± 75.3) mg/g,both P ＜ 0.01) and in the levels of cross-linked collagen as well as mRNA expressions of collagen Ⅰ and Ⅲ (P ＜ 0.01 or 0.05),but a significant increase in the content of AGE ((37.47 ± 10.65) vs.(26.39 ± 3.74) AUF/mg hydroxyproline,(47.70 ± 5.66) vs.(29.91 ± 6.50) AUF/mg hydroxyproline,both P ＜ 0.01) and MDA ((6.62 ± 0.47) vs.(4.82 ± 0.56) μmol/L,(8.63 ± 0.36) vs.(5.15 ± 0.46) μmol/L,both P＜ 0.01) in skin tissue,compared with the control mice.The level of non-cross-linked collagen in skin tissue was also lower in the diabetic mice than in the control mice on week 12 (P ＜ 0.05).Moreover,the contents of hydroxyproline and the expression levels of collagen I in skin were significantly lower (P ＜ 0.05),but the levels of AGE and MDA were significantly higher (P ＜ 0.01) in the diabetic mice on week 12 than in those on week 4.Conclusions The characteristics of collagen fibers in skin are altered in diabetic mice when compared with normal control mice,which may be associated with increased AGE content and oxidative injury in skin.

Objective:To determine whether CD46, CD55, and CD59 are differentially expressed in neoplastic and adjacent nor-mal colon tissues and to investigate their influence on clinicopathologic variables. Methods:Immunohistochemistry (a modified two-step method) was used to detect the expression of CD46, CD55, and CD59 in a tissue microarray of 121 cases of colon cancer and corre-sponding adjacent non-tumor tissues with detailed clinical information, including gender, age, differentiation, TNM classification, tu-mor location, and tumor histotype. The colon carcinoma microarray was constructed from patients' samples obtained from the Depart-ment of Gastrointestinal Surgery of Xijing Hospital of the Fourth Military Medical University between October 2004 and June 2006. The correlation between expression and clinicopathologic features was analyzed. Results:The expression levels of CD46, CD55, and CD59 were significantly higher in colon cancer tissues compared with those in normal adjacent colon tissues (P<0.001). CD46 expres-sion was not related to any specific patient characteristics, such as age, gender, grade of tumor differentiation, or TNM classification (P>0.05). The expression levels of CD55 and CD59 were correlated with the grade of colon cancer differentiation. Low levels of CD55 and CD59 were detected in cancer cells of highly differentiated cancer, whereas stronger staining for CD55 and CD59 was mainly observed in cancer cells of moderately and poorly differentiated colon cancer (P<0.05). In addition, the expression levels of CD55 and CD59 were higher in stages III and IV colon cancer than those in stages I and II according to TNM classification (P<0.05). Conclusion:CD46, CD55, and CD59 are up-regulated in colon cancer. Specifically, CD55 and CD59 are of clinical relevance to differentiation and TNM staging of colon cancer, and their expression might be closely related to clinical biological behaviors.

Objective:To analyze risk factors of breast ductal carcinoma in situ (DCIS) with microinvasion (DCIS-MI) and explore suitable axillary lymph node surgery treatment for patients with DCIS-MI. Methods:The clinical characteristics, such as age, menopausal status at diagnosis, size of breast mass, and pathology reports of 45 patients with breast DCIS or DCIS-MI treated at Jinling Hospital, Medical School of Nanjing University from February 2013 to February 2016, were retrospectively collected and analyzed statistically to deter-mine the risk factors associated with microinvasion. Results:Premenopause (P=0.006), tumor size≥3.15 cm (P=0.006), and family his-tory of malignant tumor (P=0.002) were proven risk factors of DCIS-MI. Conclusion:Patients with clinical palpable axillary mass, pre-menopause, large breast mass, and family history of malignant tumor demonstrated high possibility of DCIS-MI. Hence, sentinel lymph node biopsy should be performed. Axillary lymph node dissection is highly recommended to patients whose main symptom is palpable axillary mass.

OBJECTVE:To investigate the clinical effect and safety of pregabalin combined with gabapentin in the treatment of central pain after cerebral infarction.METHODS:One hundred and fifty patients with central pain after cerebral infarction in our hospital from Jan.2010 to Dec.2015 in our department were randomly divided into group A,B,C,with 50 cases in each group.Group A was given Pregabalin capsule 75 mg,bid combined with Gabapentin capsule 0.1 g,tid;group B was given Pregabalin capsule 75 mg,bid;group C was given Gabapentin capsule 0.1 g,tid;3 groups were treated for 4 weeks.VAS score,NRS score,PSQI and SF-36 score were observed among 3 groups before and after treatment to evaluate clinical efficacies of 3 groups;the occurrence of ADR were recorded in 3 groups.RESULTS:The clinical total response rate of group A,B,C were separately 94.00％,74.00％,70.00％.The clinical total response rate of group A was significantly better than that of group B and C,with statistical significance (P＜0.05).After treatment,VAS score of group A,B,C were separately(3.87 ± 0.74),(5.10 ± 1.26),(5.03 ± 1.23);NRS score were separately (3.91 ± 0.88),(5.29 ± 1.25),(5.37 ± 1.30);VAS score and NRS score of group A were signifi cantly lower than group B,C and before treatment,with statistical significance (P＜0.05);PSQI score of group A,B,C were separately(4.03 ± 0.85),(5.92 ± 1.16),(5.83 ± 1.11);SF-36 score were separately (372.84 ± 73.25),(348.07 ± 60.54),(345.67 ± 59.72);PSQI score and SF-36 score of group A were significantly better than group B,C and before treatment,with statistical sig nificance (P＜0.05).There was no statistical significance in the incidence of ADR among 3 groups (P＞0.05).CONCLUSIONS:Compared with pregabalin and gabapentin alone,pregabalin combined with gabapentin in the treatment of central pain after cerebral infarction can efficiently relieve the perceived pain,improve sleep quality and daily life quality and not increase the risk of ADR;therefore,drug combination plan is recommended for patient with central pain after cerebral infarction,especially with poor effect of two single drug.

Purpose To investigate the value of contrast enhanced ultrasound (CEUS) in evaluating blunt hepatic trauma and to provide experimental reference. Materials and Methods Fifteen healthy New Zealand white rabbits were randomly divided into groups A, B and C, with 5 rabbits in each group. Hepatic area of the rabbits in each group was stroked using a self-made mini striker with forces of 127.8 N, 213.0 N and 298.2 N, respectively, to establish rabbit models of blunt hepatic trauma. Gray-scale ultrasound (US) and CEUS were performed within 20 minutes before and after trauma. Results Hepatic capsular rupture was observed in 10 rabbits on US. Ill-defined hypoechoic areas in various degrees were observed. CEUS and time-signal intensity curve analysis showed that the normal hepatic tissue began to enhance after contrast agent injection, well-defined against non-enhanced hepatic lesions. The sensitivity of US and CEUS for blunt hepatic trauma were 66.7% (10/15) and 100.0% (15/15), respectively (P<0.05). The agreement with specimen was 40.0% (6/15) for US and 86.7% (13/15) for CEUS, respectively (P<0.05), with Kappa=0.815 for CEUS (P<0.001). Conclusion CEUS improves diagnostic accuracy of blunt hepatic trauma and accurately assesses the severity, which shows good agreement with gross specimen.

Objective To evaluate three-dimentional (3D) reconstruction technique and methylene blue staining in precise anatomic hepatectomy.Methods The clinical data of 12 patients with hepatocellular carcinoma who were admitted to the Chinese PLA General Hospital from February 2009 to August 2011 were retrospectively analyzed.The 3D reconstruction of the liver tumor and intrahepatic vessels were done based on the computed tomography data and magnetic resonance imaging data.The portal vein supplying the tumor and its anatomic relationship with adjacent vessels were evaluated.Precise anatomic hepatectomy was performed guided by sustained methylene blue staining.Results The accurate rate of 3D model of the portal triad was 12/12.The shape of target segments observed after methylene blue staining was consistent with the results of 3D evaluation.Two patients received hemihepatectomy,3 received lobectomy,5 received monosegmentectomy or subsegmentectomy,2 received multisegmentectomy.The mean tumor diameter,operation time,blood loss,postoperative hospital stay and complication rate were 5.6cm (2.5-16.0 cm),(150±24)minutes,(236±25)ml,(10±3)days and 2/12,respectively.After a median follow-up of 14 months,tumor recurrence was found in 2 patients,and 1 of them died of tumor progression.Conclusions The 3D reconstruction may contribute to precise evaluation of the anatomic relationship between the tumor and its adjacent vessels.The 3D technique combined with sustained methylene blue staining may significantly improve the accuracy of anatomic hepatectomy.

Objective To study the relationship between the polymorphism of interleukin-12B (IL-12B)gene and coronary heart disease.Methods We recruited 256 patients with coronary heart disease admitted to our department as the study group and 256 normal subjects as the control group.The polymorphism of IL-12B gene was detected by polymerase chain reaction and single nucleotide polymorphism.Coronary artery stenosis,visfatin,high sensitive C reactive protein and cardiac function were determined.Results The difference in rs15677380 and rs14050311 allele frequencies between the study group and the control group was significant (χ2 =6.19,7.24,P=0.045,0.021).The G allele of rs15677380 and C allele of rs14050311 were risk factors for coronary heart disease (OR=1.32,1.49).Conclusion IL-12B gene is associated with the occurrence and development of carotid atherosclerosis and participates in the development of coronary heart disease.

A rapid, sensitive and simple liquid chromatography-tandem mass spectrometric (LC-MS/MS) method was developed for the simultaneous determination of yogliptin and its metabolite in Wistar rat plasma. Linagliptin and dexamethasone were chosen as the internal standards of yogliptin and its metabolite, (R)-8-(3-hydroxypiperidine- -yl)-7-(but-2-yn-1-yl)-1-((5-fluorobenzo[d]thiazol-2-yl)methyl)-3-methyl- H-purine-2, 6 (3H, 7H)-dione, respectively. After a simple protein precipitation using acetonitrile as the precipitating solvent, both analytes and ISs were separated on a Grace Altima HP C18 column (2.1 mm x 50 mm, 5 microm) with gradient elution using methanol (containing 0.1% formic acid, 4 mmol x L(-1) ammonium acetate)-0.1% formic acid (containing 4 mmol x L(-1) ammonium acetate) as the mobile phase. A chromatographic total run time of 4.4 min was achieved. Mass spectrometric detection was conducted with electrospray ionization under positive-ion and multiple-reaction monitoring modes. Linear calibration curves for yogliptin and its metabolite were over the concentration range of 0.5 to 500 ng x mL(-1) with a lower limit of quantification of 0.5 ng x mL(-1). The intra- and inter- assay precisions were all below 14%, the accuracies were all in standard ranges. The method was used to determine the concentration of yogliptin and M1 in Wistar rat plasma after a single oral administration of yogliptin (27 mg x kg(-1)). The method was proved to be selective, sensitive and suitable for pharmacokinetic study of yogliptin and M1 in Wistar rat plasma.