OBJECTIVE: To probe into the application of antibiotics in orthopedic perioperative period. METHODS: The application of 591 patients with orthopedic disease in perioperative period from Jun. 2006 to Jun. 2008 were reviewed and analyzed. RESULTS: 591 patients were treated with antibiotics in perioperative period. 85 cases of preoperational medication accounted for 14% and 286 cases of drug combination 48%. Among 286 cases of drug combination, 263 cases of two-drug consisted of 92%, 23 cases of three-drug 8%; mean administration time was(9?2.3)d. CONCLUSION: Abuse, inappropriate use and inappropriate drug combination of antibiotics were existed, which need to be corrected.

Objective To establish a SYBR green Ⅰ real-time PCR method for detecting serum miR-21 and preliminarily explore its value in diagnosing breast cancer.Methods Total RNA was extracted from serum by Trizol reagent.Then miR-16 ( internal reference gene for miR-21 ) and miR-21 were reverse transcribed into corresponding cDNA by stem-loop RT primers.Their cDNA were amplified and detected by using SYBR green Ⅰ real-time PCR.The accuracy of assay was analyzed by signal to noise ratio (SNR) ; the specificity of assay was evaluated by melting curve; the precision of assay was assessed by R2 of standard curve and the stability of assay was calculated by intra-assay and inter-assay variation.Furthermore,the level of miR-21 and miR-16 were detected by this method among the serum samples of 33 breast cancer patients,18 benign breast disease patients and 49 healthy individuals.And the sensitivity and specificity in breast cancer diagnosis were evaluated according to cut-off value which was defined by relative expressions of miR-21 between breast cancer patients and healthy population.Results Through optimization of temperature and time in the annealing and extension stage during PCR,SNR was ≥99.36％ ; peak of melting curve was single; R2 of standard curve was 0.994 8 and Coefficient of Variance (CV) of intra-assay ＜ 1.5％,CV of inter-assay ＜4％.They indicated that this method was accurate,specific,precise and stable.When miR-16 was taken as internal reference,the relative expressions of serum miR-21 detected by SYBR green Ⅰ realtime PCR among the serum samples of breast cancer patients,benign breast disease patients and healthy population were 20.83 ± 18.18,20.86 ± 10.11 and 9.33 ±4.44,which had statistical significance among of them (x2 =16.92,P ＜ 0.001 ).There was statistical significance in healthy people vs breast cancer patients ( Z =- 2.58,P ≤ 0.01 ) and healthy people vs benign breast disease patients ( Z =- 4.42,P ≤0.01 ),but was not between breast cancer patients and benign breast cancer patients (Z =-0.51,P =0.608).When the value of 18.32 for the relative expressions of miR-21 was defined as cut-off value,the sensitivity and specificity of this method to diagnose breast cancer were 51.5％ (17/33)and 93.9％ (46/49),respectively.Conclusions A sensitive,specific and stable SYBR green Ⅰ real-time PCR for detecting serum miR-21 has been established.This method may have some diagnostic value for breast cancer.

Gastroenteropancreatic neuroendocrine neoplasms are a group of heterogeneous tumors originated from the gastrointestinal peptidergic neurons and neuroendocrine cells.Rectum is one of the major sites of gastroenteropancreatic neuroendocrine neoplasms.Although the progression of rectal neuroendocine neoplasms (rNENs) is relatively slow,metastasis could occurs and liver is the major target organ invaded by distant metastatic rNENs.Surgical operation can be used as the preferred method for the treatment of rNENs and its metastasis at present.If the clinical situations of tumors are not eligible for the operation,interventional treatment can be considered as an alternative treatment.The chemotherapies and molecule-targeted drugs for rNENs are primarily in the light of therapeutic methods for pancreatic neuroendocrine neoplasms,it may help patients achieve partial response or stable disease,and prolong patients survival time.However,there has been few clinical researches directed toward rNENs and no an extensive consensus for treatment of rNENs up to now,and more evidences of evidence-based medicine should be necessitated.This paper has discussed and summarized treatment progression of rNENs and its metastasis.

Pancreatic neoplasm is one of the most commonly-appeared digestive tumors and has been well-recognized as the poor diseases which have the difficulties in diagnosis,treatment and prognosis estimation.Recently,the detection of circulating tumor cells (CTCs) has been a pretty highlight of the research on detecting tumor cells in peripheral blood,and furthermore,the clinical value in the diagnosis,treatment and prognosis prediction has already been verified through a large amount of samples analyses in various kinds of tumor diseases.This paper aims to review and conclude the techniques of CTCs enrichment,detection and clinical implications in pancreatic neoplasms.In addition,the existing papers have been summarized and prospect of application of CTCs is also presented.

Objective To investigate the hemostatic and coagulative variations during orthotopic liver transplantation (OLT).Method The blood platelet count, coagulant and anti-coagulant functions were assessed pre- and intra-operation of OLT.Results During the operation, activated partial thromboplastin time (aPTT) and prothrombin time (PT) were prolonged, platelet count (PLT), activities of most of the coagulation factors and levels of antithrombin (AT), plasminogen (PLG), plasminogen activator inhibitor-1 (PAI-1) and ?2 -antiplasmin (?2-AP) were reduced, while the levels of tissue plasminogen activator (t-PA), plasmin-?2-antiplasmin complex (PAP) and thrombin-antithrombin complex (TAT) were increased. The variations in the neohepatic phase were more significant than that in the pre-operation phase. Conclusion In the entire process of OLT operation, the coagulant and anti-coagulant functions were decreased, and the fibrinolytic functions were sthenic in the anhepatic phase and the neohepatic phase.

Objective To explore the practical value of liver transient elastography technique in the detection of chronic hepatitis B fibrosis.Methods The liver biopsy and transient elastography technique was detected in 106 patients with chronic hepatitis B.Observation index included:alanine aminotransferase (ALT),aspartate aminotransferase (AST),total bilirubin (TBIL),albumin (ALB),platelet (PLT),statistial analysis was performed in liver biopsy and fibroscan results by SPSS17.0 software.Results Liver stiffness measurement (LSM) was positively correlated with ALT,AST and TBIL,and which was negatively correlated with ALB,PLT,and which was positively correlated with liver fibrosis stage.The area under the ROC curve by the LSM prediction of liver fibrosis S4,S3,S2 stage were 0.869 (95％ CI 0.809-0.918),0.841 (95％ CI 0.810-0.901),0.806 (95％ CI 0.747-0.866),the corresponding critical value was 22.9,14.8,10.2 kPa.Conclusion There is a good correlation between fibroscan and liver fibrosis,LSM ＞ 10.2 kPa can be considered in patients with antiviral therapy.

Objective To prepare mouse polyclonal antibody against human Nanog by genetic immunization and to identify this antibody by Western blot and immunofluorescence. Method The antigenicity fragment (A16-V101) of human Nanog (hNanog) was chosen by analysis of Accelrys software, and its cDNA (258bp) was amplified from plasmid containing full-length cDNA of hNanog, then it was cloned into pBQAP-TT to construct recombinant plasmid pBQAP-TT-hNanog for genetic immunization. Mice were immunized with this recombinant plasmid and two other adjuvant plasmids-pCMVi-GMCSF and pCMVi-FIT3L, which help to enhance the antibody's generation. After 12 weeks, we obtained mouse anti-hNanog antibody from mice blood serum. The antibody titer was determined by enzyme-linked immunoadsordent assay (ELISA), and its specificity was identified by Western blot in human renal protein. Using this antibody, we detected hNanog expression in HKC cells of hNanog-AAV2 transfection. Results Recombinant plasmid pBQAP-TT-hNanog for genetic immunization was confirmed to be correct by restriction digestion and sequencing. The result of ELISA showed that the antibody titer was 1∶3 200. This antibody recognized a band of 34kD hNanog protein in human renal protein by Western blot. Immunofluorescence showed that Nanog protein was mainly located in the nuclei in hNanog transgene HKC cells. Conclusion Genetic immunization can offer mouse anti-hNanog polyclonal antibody of high titer and high specificity.

Objective To analyze the clinicopathological characteristics and the related factors influencing the prognosis of pancreatic neuroendocrine carcinoma.Methods The clinicpathological and follow-up data of 21 patients with pancreatic neuroendocrine carcinoma admitted between April 2000 and August 2013 in Zhongshan Hospital of Fudan University were analyzed retrospectively.The influence facts on patients' prognosis were analyzed statistically.Results Univariate analysis showed that age,stage,tumor size,degree of tumor differentiation and lymph node metastasis were factors separately influenced patients' survival.Multivariate analysis (Cox regression) revealed that age (P =0.019) was the only independent factor affecting the prognosis.Conclusions Patients with pancreatic neuroendocrine carcinoma were mostly non-functional and presented no specific clinical features.Tumors tended to metastasize and the prognosis was poor.Age was the independent factor affecting the prognosis.

Objective The aim of this study was to explore MMP-2 and TIMP-3 expression in the gastric cardia carcinoma and with the relationship between the pathological stages.Methods Gastric cardia carcinoma surgical resection speciments of 115 cases were for the trial group.paraneoplastic gastric noncancerous tissues of 40 cases (the distance to cancer border>5 cm) as normal controls.The expression of MMP-2 and TIMP-3 was detected by two-step immunohistochemical method.The relationship of MMP-2 and TIMP-3 between clinicopathological stages was analyze. Results MMP-2 in carcinoma of the gastric cardia and the paraneoplastic tissues of organizations positive expression rates were 57.4%(66/115) and 40.0%(16/40) (P<0.05).MMP-2 expression was positively correlated with tumor size,tumor invasion depth,lymphatic metastasis and stage of tumor, which was negatively correlated with the prognosis of gastric cardia carcinoma (P<0.05).TIMP-3 in carcinoma of the gastric cardia and the paraneoplastic tissues of organizations positive expression rates were 48.7%(56/115) and 95.0%(38/40) (P<0.05).TIMP-3 expression was negatively correlated with tumor size,tumor invasion depth,lymphatic metastasis and stage of tumor, which was positively correlated with the prognosis of gastric cardia carcinoma. Conclusion The over expression of MMP-2 and the lost of TIMP-3 may play an important role in the invasion and metastasis of gastric cardia carcinoma.Combined MMP-2 and TIMP-3 detection may help to judge gastric cardia carcinoma development and to assess the prognosis of patients.

Objective To study the effects of enteral immunonutrition on immune function in patients with multiple trauma. Method A total of 32 patients with multiple trauma admitted to general ICU in ChangZheng hospital, Shanghai from March 2007 to May 2008 were randomly (random number) divided into two groups,enteral immunonutrition group (EIN group, n = 16)and the general formula enteral nutrition group (EN group, n = 16).Two groups of patients on the third day after injury began to accept the EIN and the general formula EN which respectively support at least 14 days. The patients peripheral blood lymphocyte count (TLC), immunoglobulin (IgG,IgM, IgA) ,and T-lymphocyte subsets (CD3, CD4, CD8, CD4/CD8) were detected on the day of the group formation, the seventh and the fortteenth day of the nutritional support. Results TLC, IgG, IgM, IgA, CD4 and CD4/CD8 ratio were significantly higher in EIN Group on the 7th and 14th day than that on the first day ( P ＜0.05), and extended with the EIN and sustained increase. All immunization indicators of EN group on the 7th day were no significant changes compared with the day of the group formation, and on the 14 th day TLC, IgG, IgM,IgA, CD4 and CD4/CD8 ratio were significantly higher than that on the day of the group formation ( P ＜ 0.05),but were significantly lower than that in the EIN group on the 14th day ( P ＜ 0.05). Conclusions Compared with the general formula EN, EIN is more helpful to restore and increase the humoral and cellular immune function in early post-multitraumatic phase.