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.

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.

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 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 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 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.

ObjectiveSmall interfering RNA (siRNA) was used to silence the fibroblast activation protein4 (FAP) expression of mouse pancreatic cancer related fibroblast cells (mPCa-FCs-1212),and to observe the effects of mPCa-FCs-1212 silencing FAP gene on mouse pancreatic cancer cells (mPCa-1212) proliferation and apoptosis.MethodsThe small interfering RNA targeting FAP gene was designed; the recombinant siRNA plasmid siFAP and control plasmid siMOCK was constructed,which were transfected into mPCa-FCs-1212,respectively.The FAP mRNA and protein expression in transfected cells were examined by real-time PCR and Western blotting.The mPCa-1212 and transfected mPCa-FCs-1212 were co-cultured with a 1:1 ratio in vitro.The growth inhibitory rates and apoptosis rates of mPCa-1212 were detected by MTT assay and Annexin V-FTTC/PI staining and FCM assay.ResultsThe mRNA and protein expressions of FAP in siFAP transfected mPCa-FCs-1212 were significantly down-regulated when compared with that in siMOCK transfected mPCa-FCs-1212[0.584 ±0.029vs.1.052±0.281,P=0.0213; (27.18±3.23)％ vs.(61.58±4.72)％,P=0.0317].The mPCa-1212 was co-cultured with the mPCa-FCs-1212 transfected with siFAP or siMOCK for 3 d,and the inhibitory rates of mPCa-1212 were (23.02 ±3.32)％ and (1.11 ±0.23)％,and the apoptosis rates were (42.31 ±5.34)％ and (7.38 ± 2.09)％,the difference between the two groups was statistically significant (P =0.000).ConclusionsmPCa-FCs-1212 silencing FAP gene can inhibit the proliferation of mPCa-1212 in vitro and induce cell apoptosis,and may be a potential new approach to gene therapy.

Objective To investigate the relationship between the changes of portal pressure gradient after selective devascularization with postoperative complications and recurrent bleeding of gastroesophageal varix in patients of portal hypertension. Methods The clinical data of 135 cases of portal hypertension undergoing selective devascularization was collected. Portal pressure gradient was measured before splenectomy and after selective devascularization, and was analyzed against postoperative complications and recurrent bleeding. Results In this study, 135 patients of portal hypertension underwent selective devascularization, two cases died during perioperative period ( 1.5% ). Postoperatively patients were divided into three groups based on PPG ＜ 12 mm Hg after selective devascularization (62 cases), HVPG ≥ 12 mm Hg but a more than 20% of decrease off the pre-splenectomy baseline (41 cases) and HVPG ≥12 mm Hg with less than 20% of decrease from the baseline (32 cases). The postoperative complications between the three groups were of no significant difference ( P ＞ 0. 05 ). The 1,2,3 year cumulative rate of no variceal rebleeding of the three groups were 100% vs. 100% vs. 95%; 100%vs. 97% vs. 90%; and 100% vs. 93% vs. 87% (x2 =6. 859, P = 0. 032). COX regression analysis indicated portal vein pressure gradient was an independent prognostic factor of variceal bleeding recurrence (P=0.002). 1,2,3 year cumulative survival rates of the three groups were 100% vs. 100% vs. 94%; 98% vs. 95% vs. 92%; 97% vs. 93% vs. 88%, there were no significant difference among the three groups ( x2 = 2. 917, P = 0. 233 ). Conclusions The decrease in the PPG after selective devascularization is a predictor for the risk of rebleeding but not for survival after selective devascularization.