Objective　To evaluate the efficacy and safty of pegging hydroxyapatite orbital implants.Methods　Fourty-five patients who received drilling and pegging of hydroxyapatite orbital implants were studied retrospectively, and all patients used the peg and sleeve system followed-up 2 to 24 months.Results　The rate of success of the first operations is 82.2%. Complications associated with pegging included peg hole drilled on an angle, drilling off-center, drilled hole shallow, granuloma and infection, total complications occured in 22.2% in the review, most that are of a minor nature can be cured.Conclusions　The hydroxyapatite orbital implants can be directly coupled to the prosthesis through a peg system, allowing a wide range of prosthesis movement and giving a more lifelike quality to the prosthesis. When and how to drill and exactly hole on implant is the key for pegging.

Objective To explore the effects of DNMT3b on the expression and methylation sta-tus of the promoter region of DLC-1 in human hepatocellular carcinoma cell line. Methods The SMMC-7721 cell line was divided into 2 groups. The cell line in the experimental group was transfect-ed with DNMT3b siRNA, while that in the control group was transfected with control siRNA. West-ern blot was used to detect the expression of DNMT3b and DLC-1 and MSP was employed to examine the methylation status of the promoter region of DLC-1. Results The expression of DNMT3b was significantly higher in the experimental group than in the control group, while the expression of DLC-1 was just opposite. There was no significant difference in the methylation status of the promoter re-gion of DLC-1 between the 2 groups and both were methylated. Conelnsion The inhibition of expression of DNMT3b by siRNA method can enhance the expression level of DLC-1, and the methylation status of the promoter region of DLC-1 does not change at the same time. When affecting the expression of DLC-1, DN-MT3b might not play the role of methyhransferase, but can act as a transcriptional regulatory factor.

Objective To study the effects of histone deacertylase inhibitor (TSA) on promoter methylation and expression of E-cadherin gene in a hepatocellular carcinoma cell line SMMC7721. Methods Hepatocellular carcinoma cell line SMMC-7721 was treated with TSA (300 nm/L), MTT method was used to investigate the growth inhibition ratio, TUNNL was conducted to measure the apoptosis ratio, methylation-specific PCR (MSP) was employed to detect changes in the CpG island methylation of E-cad promoter region, Western blot technique was used to detect the expression of E-cad gene and DNMT3b before and after TSA treatment, respectively. Results TSA decreases the SMMC-7721 cell viability and induces apoptosis, the growth inhibition ratio was 21.85% compared with control group. The apoptosis ratio of control group was (4.69±0.56)% ,the apoptosis ratio of TSA treatment group was (14.94±0.91)%. The apoptosis ratio of TSA treatment group was significantly higher than that of control group(P = 0.000). Before treated with TSA, the CpG island of E-cad promoter region was methylated, and the expression of E-cad was negative. TSA treatment induces demethylation of the CpG island in E-cad promoter region, causes the re-expression of E-cad. TSA reduces the expression of DNMT3b. Conclusions TSA decreases the SMMC-7721 cell viability and induces apoptosis, reverses the methylation status of E-cad promoter region, and resumes E-cad gene expression. TSA may induce demethylation through down-regulating the expression of DNMT3b.

Objective To evaluate the differences of the thinnest-point corneal thickness (TCT) decrease after three different corneal crosslinking (CXL) protocols for progressive keratoconus.Methyds Retrospective clinical case study.From August 2010 to November 2015,consecutive 85 patients (110 eyes) with progressive keratoconus were enrolled and treated with CXL in Department of Opthalmology,Navy General Hospital.21 patients of 25 eyes underwent standard epithelium-off corneal crosslinking (S-CXL),14 patients of 22 eyes underwent 1 g · L-1 riboflavin-sodium lactate Ringer's solution iontophoresis-assisted CXL (I-CXLa),and 50 patients of 63 eyes underwent 0.1％ riboflavin-distilled water solution I-CXLb.Preoperative and postoperative TCT were measured by ALLEGRO oculyzer.The differences of TCT decrease after treatment were compared among the three CXL protocols.Results The differences of TCT from baseline after 3 months,6 months and 12 months in the S-CXL group were (-14.93 ±27.16) μm,(-31.94 ±22.89) μm,(-27.71 ±26.01) μm,respectively,the I-CXLa group were (-20.14 ± 19.09) μm,(-10.10 ± 24.28) μm,(-7.11 ± 22.26)μm,respectively,the I-CXLb group were (-28.08 ± 26.14) μm,(-21.08 ± 25.62) μm,(-15.91 ± 19.19)μm,respectively.Three months after treatment,the differences of TCT decrease in the three groups was not statistically significant (P =0.188);Six and 12 months after treatment,the differences between S-CXL and I-CXLa were statistically significant (all P ＜0.05),but the differences between S-CXL and I-CXLb,between I-CXLb and I-CXLa showed no significant difference (all P ＞ 0.05).Conclusion Six and 12 months after treatment,TCT decrease is related to the CXL protocol.TCT decrease degree may reflect the intensity of crossinking.TCT decrease in I-CXLb is smaller than that in S-CXL,but there is no statistical difference.

Objective　To evaluate the diagnosis and surgical treatment of hilar cholangiocarcinoma(H-CC). Methods　Retrospective analysis was made on the clinical feature, surgical treatment and the effect on 73 patients with H-CC. Results　Diagnosis was made in all of the patients preoperatively and the correct diagnostic rate of BUS was 69.9%. In the treatment, radical resection was performed on 15 patients with good results in a short-term period. Of the 43 patients who underwent biliary tract internal drainage or exterrnal drainage, 37 patients had good results in a short-term period, while 6 died after operation. Laparotomy or hepatic artery cannulization with chemotherapy was performed on 15 patients and no change occurred in a short-term period after operation. In 15 cases subjected to radical resection, 11 cases were followed up. The 1,3-year survival rates was 90.9%, 20.0% respectively, but none of the patients survived for over 5 years. In patients undergoing other operations, none survived more than 9 months. Conclusions　It's still difficult to mak early diagnosis of H-CC, which mainly depends on imaging technics. The BUS should be choiced first. Radical resection rate is still low nowadays. The lobus quadratus resection is helpful to select the operation.

Objective To investigate the expression of Golgi protein?73(GP73) and Ki?67 antigen in gallbladder carcinoma ,and to analyze their correlations with proliferation ,invasion ,and prognosis of gallbladder carcinoma. Methods Streptavid?in?peroxidase (SP) immunohistochemistry was used to detect the expression of GP73 and Ki?67 in surgically resected specimens of 58 gallbladder carcinomas ,15 gallbladder adenomas and 15 gallbladder polyps samples . Results The positive rates of GP73 and Ki?67 protein in gallbladder carcinomas were 72.4% and67.24%,respectively ,which wer significantly higher than those in gallbladder adenomas(GP73:40.0%,Ki?67:26.7%,P<0.05)and in gallbladder polyps(GP73:13.3%,Ki?67:25.0%,P<0.05).The expression of GP73 was positively correlated with tumor differentiation ,Nevin staging and lymph node metastasis(P < 0.05), and the expression of GP73 was positively correlated with tumor differentiation ,Nevin staging(P < 0.05). GP73 expression was positively correlated with Ki?67 expression in gallbladder carcinoma (r = 0.473 ,P = 0.000). Patients with negative expression of GP73 and Ki?67 had longer survival time than those with positive expression of GP73 and Ki?67. Conclusion The expression of GP73 and Ki?67 was associated with proliferation ,invasion of gallbladder carcinoma. The combined detection of GP73 and Ki?67 is conducive to judging the progress and prognosis of the gallbladder carcinoma .

Objective To explore the expression of PIWIL1 protein and DICER enzyme in hepa tocellular carcinoma (HCC) and their significance.Methods Immunohistochemical method was used to detect the expression of PIWIL1 and DICER in 47 cases of HCC and the adjacent HCC tissues.Western blot method was used to detect the expression of PIWIL1 and DICER in 31 cases of fresh HCC tissues and their adjacent HCC tissues.The relationship between PIWIL1 and DICER and their relationships were analysed with clinical features.12 cases of normal liver tissues were used as control group.Results The expression of PIWIL1 was high in HCC but low in normal liver tissues (P＜ 0.05).The expression of DICER was high in normal liver tissues but low in HCC (P＜0.05).The expression of PIWIL1 was positively correlated with invasion to adjacent tissues and histological differentiation (P＜0.05).The expression of DICER was negatively correlated with invasion to the adjacent tissues and histological differentiation (P＜0.05).There was a negative correlation between PIWIL1 and DICER (P＜ 0.05).Conclusions High expression of PIWIL1 and low/missing expression of DICER was related to pathological differentiation and invasion of adjacent tissues.

Objective To investigate the effect of LI-cadherin- SiRNA protein on the growth and metastatic potentials of transplanted human hepatocellular carcinoma cell lines (Hep3B) in nude mice.Methods We transfected LI-cadherin- SiRNA to Hep3B cells,Hep3B cell suspension (transfected or control ) was injected subcapsullaryly into the spleen of nude mice,hepatic metastasis was observed by naked eye and immunohistochemistry.In addition,Western-blot was used to detect the level of LI-cadherin in different metastatic site.Results (1) Hep3B was green after successful transfect interference vector under fluorescence inverted microscope,in this study,the transfect rate was 80％.(2) Hep3B liver metastasis model in nude mice was established.The metastasis rates in the empty plasmid carrying group,the control group and the SiRNA transfect group were 50％,60％ and 80％,respectively.The number of metastasis caner nodules in the SiRNA transfect group was 26,significantly higher than other two groups.(3) The level of protein expression for LI-cadherin in the SiRNA transfect group is significantly lower than the control group and the empty plasmid carrying group.Conclusions LI-cadherin is crucial and important for the adhension capability of HCC in its migration.SiRNA transfected LI-cadherin increases the metastasis in a nude mouse model inoculated with human hepatocellular carcinoma cell lines.

Objective To investigate the diagnostic value of macrophage migration inhibitory factor (MIF) for hepatocellular carcinoma (HCC).MethodsThe research was divided into 2 parts,including testing research and confirmatory research.The clinical data of 269 patients with HCC ( group A) and 390 individuals (including 135 patients with hepatic cirrhosis,106 with benign hepatic diseases and 149 healthy individuals,control group A) who were admitted to the Zhongshan Hospital of Fudan University from January to May,2004,and 173 patients with hepatic cancer (group B) and 257 individuals (including 86 patients with hepatic cirrhosis,79 with benign hepatic diseases and 92 healthy individuals,control group B ) who were admitted from August to December,2004,and 80 patients with HCC who received radical hepatic resection in January 2005 were retrospectively analyzed.Samples of plasma of patients in the group A and individuals in the control group A were collected before operation.Samples of plasma of patients received radical hepatic resection were collected preoperatively and at postoperative day 3,7 and 30.HCC and adjacent issues of patients in the group A were collected.The levels of MIF in the plasma and tissues were detected by enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry,respectively.Non-normal distribution data were described as M( QR).Differences between the groups were analyzed by using the Mann-Whitney U test,and the relationship between the levels of MIF in the plasma and tissues was detected by the Spearman correlation coefficient.The diagnostic value of MIF was analyzed by the ROC curve.ResultsThe levels of MIF in the plasma of patients in the group A and individuals in the control group A were 85.7 μg/L (58.8 μg/L) and 15.5 μg/L(31.6 μg/L),respectively.The levels of MIF in the plasma of the patients with hepatic cirrhosis,benign hepatic diseases and healthy individuals were 24.9 μg/L (12.6 μg/L),12.5 μg/L(7.3 μg/L) and 13.2 μg/L (7.7 μg/L),respectively.There was a significant difference in the level of MIF between the group A and the control group A (F =54.235,P ＜ 0.05 ).The area under the ROC curve reached peak when the level of MIF in the plasma was 35.3μg/L.Compared with the control group B,the vdues of AUC,sensitivity and specificity were 92.1％,90.7％ and 93.4％ in the group B.The levels of MIF of the patients with HCC before operation and at 3,7,and 30 days after operation were 81.0 μg/L(54.0 μg/L),76.1 μg/L(47.5 μg/L),50.9 μg/L (40.7 μg/L) and 18.7 μg/L ( 15.1 μg/L),respectively.The levels of MIF decreased with time passed by,and were back to normal at 30 days after the operation.The median expressions of MIF in the HCC and adjacent issues were 0.083 and 0.007,respectively,with a significant difference ( U =3.975,P ＜ 0.05).The expression of MIF in the plasma was positively correlated with its expression in the HCC tissue ( r =0.759,P ＜ 0.05 ).ConclusionMIF plays an important role in the genesis and development of HCC and has potential to be one of the molecular markers for the diagnosis of HCC.

Objective By analyzing the expression of cluster of differentiation 74 (CD74) in hepatocellular carcinoma (HCC) and HCC cell lines,the correlation between the level of CD74 expression and the patients' prognosis was investigated.MethodsThe expression of CD74 in high metastatic potential HCC cell lines(MHCC-LM3,MHCC-97H),low metastatic potential HCC cell line( MHCC-97L),and no metastatic potential HCC cell line(Hep-G2) were estimated by Western blot.The paraffin embedded tissues which include intra-tumor and paratumor tissues were collected from 320 patients who had received HCC curative surgical resection and 5 normal liver tissues from the donors of liver tranplantation.The high density tissue micro-array was made of these specimens. Immunol-histochemistry was applied to discover the different levels of CD74 in tumor,paratumor and normal liver tissues.Survival curves were generated by the Kaplan-Meier method and verified by the Logrank test.Cox proportional hazards regression analysis was applied to estimate the prognostic factors in multivariate analysis.ResultsThe expression level of CD74 was significantly higher in low metastatic potential and no metastatic potential HCC cell lines (MHCC-97L 1.224 ±0.014,Hep-G2 1.374 ±0.006) than that in high metastatic potential ones( MHCC-LM3 0.622 ±0.078,MHCC-97H 0.732 ± 0.083 ).Significant differences can be found between the groups (t =- 13.308,- 16.849,- 10.177,- 13.436,- 17.057; P ＜0.01 ).Meanwhile,in tumor tissues,the CD74 was expressed positively in 221 patients and negatively in 99 patients.But CD74 was expressed slightly in paratumor and negatively in 5 normal liver tissues.There's no significant differences between the groups categorization according to age,HBsAg,cirrhosis,AFP level,tumor number,tumor size,tumor capsule,blood vessel invasion,Edmondson Grades and tumor nodes metastasis classification (TNM) stages (x2 =0.053,0.141,1.200,0.000,0.277,1.975,0.263,1.044,0.000,0.433 ; P ＞ 0.05 ),except gender (x2 =3.954,P ＜ 0.05).Kaplan-Meier method showed that patients with positively expression of CD74 had better prognosis than others (x2 =5.620,P ＜ 0.05 ).Cox proportional hazards regression analysis showed that CD74 was a significant and independent prognostic factor of survival [ hazard ratio (HR) =0.721,95％confidence interval (CI) =0.522 - 0.996,P ＜ 0.05 ].Conclusion The expression of CD74 in hepatocellular carcinoma could be a biomarker of the prognosis and there's some potential correlation with cancer cell apoptosis.