[Objective] To recommend an ideal method for extracting Ⅰ-collagen from cortical bone.[Method]The cortical bone of pig was splitted into small pieces after the soft tissues were cleaned up.The bone pieces were gradually dehydrated in alcohol,defatted in aether,decalcificated in hydrochloric acid,redefatted in chloroform:methanol(1:1,v/v)and became soft.The soft pieces were disintegrated into demineralized bone matrix(DBM)powder in a high speed mill.The osseins were extracted respectively by enhanced pepsin digestion method or alkali-solution method after the DBM powder was treated with desolving,centrifuging,dialyzing and lyophilization.The product got by enhanced pepsin digestion method was further confirmed.The extraction rate and appearance,viscosity and solidification of both products were analyzed and compared.[Result]The collagen produced by extraction of enhanced pepsin digestion method was confirmed to be I-collagen by analyzing amino-acid composition,protein electrophoresis,relative moleculas weight and max wavelength about light absorption.The extraction rate of Ⅰ-collagen by enhanced pepsin digestion method was(94.0?14.96)% as comparision with by alkali-solution method(57.8?4.96)% and the viscosity test of acetic and solution at the concentration 0.03%(w/v)was 3.71 and 2.81 respectively.At the condition of 37?,pH 7.35~7.45,the collagen solution extracted by enhanced pepsin digestion method solidified and changed into a glue 10 minutes later,while the product solution by extraction with alkali-solution was still like a sticky liquid.[Conclusion]Ossein extracted from cortical bone with enhanced pepsin digestion method is a realⅠ-collagen.Compared with alkeli-solution extraction method,the enhanced pepsin digestion extraction method has advantages of higher extraction rate,better purity,better viscosity and fine solidification of the product,and is a choice to prepare the Ⅰ-collagen from cortical bone.

[Objective]To investigate the diagnosis,surgical procedure and clinical effect for the subdural extramedullary tumor. [Methods]Forty-five patients (28 males and 17 females,age ranged from 24-71 years with an average of 45 years ) were treated with tumour extirpation. MRI showed subdural extramedullary tumor before surgery. The spine was exposed through a standard posterior approach,and the dura matter was opened after lamninectomy. The tumor was separated and removed from the dura matter. Pedicle screw fixation was performed to restore the stability of spine. [Results]The patients were followed up from 6 to 42 months with an average of 26.2 months. The postoperative recovery of neurological function was measured with Otani scores. The clinical result was excellent in 11 patients,good in 18 and fair in 12,resulting in 64.44% of good to excellent rate. The good to excellent rate had an advantage in patients (84.62%) operated on within one month after first visit over that of patients (64.29%) who had a misdiagnosis for 1~6 months and the patients (50.00%) for more than 6 months.[Conclusion]Removal is the method of choice for patients with subdural extramedullary tumor. Early diagnosis and removal are key factors to get a good clinical effect.

Objective To discuss the expansion model of the eyeball and investigate the morphologic characteristics of high-my- opic eyeball through the dimensional measurement in high-myopia and emmetropia by MRI. Design Case controll study. Participants Thirty-two emmetropes (60 eyes) and 33 high myopes (60 eyes) were enrolled, without eye diseases and history of ocular surgery or in- jury. Methods 60 high-myopic eyes and 60 emmetropic eyes were measured with MRI (I.5T,PHILIPS) to get the data of three inner ocular dimensions, intraocular volume and the volume of different parts. Main Outcome Measures Three dimensions and volumes of eyeballs. Results The average value of axial (28.16?2.80 mm), horizontal (22.87+1.23 mm) and vertical length (23.40?0.99 ram) of high-myopic eyes were much bigger than those of emmetropic eyes(P=0.000), especially the axial length( with difference of 5.38 mm); The axial length was correlated with refractive error (0.36 mm/D,r~2=0.88, P=0.000). The average value of the whole ocular volume (7. 46?0.89 ml) and vitreous volume(6.90?0.8 ml) of the high myopic eyes were bigger than those of emmetropic eyes(P=0.000), while ante- rior segment volume and lens volume were about the same as that of emmetropic eyes (P=0.220, P=0.630). Conclusions The three di- mensions of high-myopic eyes were significantly longer than that of emmetropic eyes. In high myopes, the increased vitreous volume lead to the increase of the whole ocular volume. There may be two models in the ocular expansion of high myopia: global expansion and axial elongation expansion. More serious refractive error cause more obvious expansion in axial elongation.

Purpose:To study the relationship between the expression of CD44v6 with the clinicopathological charac- teristics and the prognosis in primary gastric carcinoma patients.Methods:A total of 188 paraffin-embedded gastric carcino- mas and 42 non-carcinomatous gastric mucosae was stained with the monoclonal antibodies CD44v6 using the EnVision~(TM) method.Results:The expression level of CD44v6 were significantly higher in the tumors (67.6%) than in the non-carcino- matous gastric mucosae (9.5%) (P

Objective To observe the effect of isolating and harvesting the chondrocytes from rabbits rib cartilage with the method of three-step enzymatic digestion, and the biological characteristic of the isolated chondrocytes during cultivation in vitro to evaluate their biological activity. Methods The method of three-step enzymatic digestion was designed that the rib cartilage was digested one by one with 1 g/L trypsin and 1 g/L EDTA, 1 g/L hyaluronidase and 2 g/L collagenaseⅠ in the culture medium to isolate chondrocytes. The harvesting and viability rate of the primary chondrocytes were detected. During the passage cultivation in vitro, the changes of the chondrocyte shape and growth were observed, and the changes of the collagen typeⅠ and Ⅱ and aggrecan in the extracellular matrix were detected. Results ① The extracellular matrix of rib cartilage was completely dissolved by the three-step enzymatic digestion, and the chondrocytes were completely isolated from the solid matrix. The number of the harvested chondrocytes from every gram of wet cartilage was (4 295.7)?10~(4) on average,and their viability rate was 97.2% on average. ②The primary and first passage chondrocytes had triangle or multi-angle shape, and became elliptic shape at the growing confluence with the positive immunohistochemical staining of collagen type Ⅱ and the strong heterochromia to toluidine blue. The content of sulfate glycosaminoglycans(GAG) in the extracellular matrix of the primary passage cells was (80.61?11.40) ?g/cm~(2). The chondrocytes after the third passage gradually became spindle shape with the negative staining of collagen typeⅡ and the weak heterochromia to toluidine blue. The content of sulfate GAG of the fourth passage cells was (44.74?10.18) ?g/cm~(2). Conclusion ① The method of three-step enzymatic digestion can make the extracellular matix of rib cartilage to be completely degraded, and has advantages of the high efficiency of harvesting primary chondrocytes with high cellular viability rate and simple manipulation. ②The primary and first passage chondrocytes have fine biological activity.

BACKGROUND: In the management of critically ill patients, the assessment of volume responsiveness and the decision to administer a fluid bolus constitute a common dilemma for physicians. Static indices of cardiac preload are poor predictors of volume responsiveness. Passive leg raising (PLR) mimics an endogenous volume expansion (VE) that can be used to predict fluid responsiveness. This study was to assess the changes in stroke volume index (SVI) induced by PLR as an indicator of fluid responsiveness in mechanically ventilated patients with severe sepsis. METHODS: This was a prospective study. Thirty-two mechanically ventilated patients with severe sepsis were admitted for VE in ICU of the First Affiliated Hospital, Zhejiang University School of Medicine and Ningbo Medical Treatment Center Lihuili Hospital from May 2010 to December 2011. Patients with non-sinus rhythm or arrhythmia, parturients, and amputation of the lower limbs were excluded. Measurements of SVI were obtained in a semi-recumbent position (baseline) and during PLR by the technique of pulse indicator continuous cardiac output (PiCCO) system prior to VE. Measurements were repeated after VE (500 mL 6％ hydroxyethyl starch infusion within 30 minutes) to classify patients as either volume responders or non-responders based on their changes in stroke volume index (ΔSVI) over 15％. Heart rate (HR), systolic artery blood pressure (ABPs), diastolic artery blood pressure (ABPd), mean arterial blood pressure (ABPm), mean central venous pressure (CVPm) and cardiac index (CI) were compared between the two groups. The changes of ABPs, ABPm, CVPm, and SVI after PLR and VE were compared with the indices at the baseline. The ROC curve was drawn to evaluate the value of ΔSVI and the change of CVPm (ΔCVPm) in predicting volume responsiveness. SPSS 17.0 software was used for statistical analysis. RESULTS: Among the 32 patients, 22 were responders and 10 were non-responders. After PLR among the responders, some hemodynamic variables (including ABPs, ABPd, ABPm and CVPm) were significantly elevated (101.2±17.6 vs.118.6±23.7,P=0.03; 52.8±10.7 vs. 64.8±10.7,P=0.006; 68.3±11.7 vs. 81.9±14.4,P=0.008; 6.8±3.2 vs. 11.9±4.0,P=0.001). After PLR, the area under curve (AUC) and the ROC curve of ΔSVI and ΔCVPm for predicting the responsiveness after VE were 0.882±0.061 (95％CI 0.759–1.000) and 0.805±0.079 (95％CI 0.650–0.959) when the cut-off levels of ΔSVI and ΔCVPm were 8.8％ and 12.7％, the sensitivities were 72.7％ and 72.7％, and the specificities were 80％ and 80％. CONCLUSION: Changes in ΔSVI and ΔCVPm induced by PLR are accurate indices for predicting fluid responsiveness in mechanically ventilated patients with severe sepsis.

Objective To study the value of PSADT in predicting the prognosis and the possibility of disease progression for patients with prostate cancer after MAB therapy.Methods Based on the retrospective review of the history and the follow-up of 159 prostate cancer patients,who received MAB therapy in our department from January 1994 to December 2010,PSADT values were calculated and survival analysis was performed.The ages at diagnosis ranged from 54 to 90 years with a median of 74 years.The pretreatment PSA value ranged from 2.6 to 275.0 μg/L with a median of 46.8 μg/L.The patients of Gleason score ≤6,7 and ≥8 constituted 27.7％,42.1％ and 25.2％,respectively.Only 26.4％ of the patients were staged as T1N0M0-T2N0M0 and the others had locally advanced disease or metastasis.A multivariate analysis with a Cox's proportional hazard model was used and the disease progression rates in different PSADT groups were also compared.Chi-square test and Log-rank test were applied in statistic analysis.Results The 159 patients received follow-up with a median period of 28 months (6-126 m).The median PSADT of these 159 patients was 5.7 months (0.5-21.0 m).The 3-year and 5-year survival for the 71 patients,whose PSADT were not less than 6 months,were 89.4％ and 47.6％ respectively,compared with 49.8％ and 30.6％ for the other 88 patients whose PSADT were less than 6 months.The survivals were significantly different between the two groups (P ＜ 0.01).It was confirmed by a further multivariate analysis with a Cox' s proportional hazard model that PSADT was one of the predictive factors of the prognosis of these prostate cancer patients with a hazard ratio of 2.6 (P ＜ 0.01).Moreover,disease progression were found in 19.7％ of the PSADT≥6 m group during the follow-up compared with 63.6％ in the PSADT ＜6 m group.The disease progression rates were also significantly different (P ＜ 0.0 l).Conclusions PSADT can be used to predict the prognosis of patients with prostate cancer after the MAB therapy.The survival for the patients,whose PSADT are not less than 6 months,is higher than those whose PSADT less than 6 months.Meanwhile,PSADT can predict the possibility of disease progression after MAB treatment.

Objective To analyze the risk factors for postoperative stricture after endoscopic submucosal dissection (ESD) for early stage esophageal cancer.Methods The data of 362 patients with early esophageal cancer treated by ESD from January 2007 to February 2012 were reviewed to investigate the risk factors of postoperative stricture.Results Esophageal stricture after ESD occurred in 42 patients (11.6％)with a mean time from ESD to stricture of (58.5 ± 12.3) days.The rates of mild,median and severe stricture were 16.7％ (7/42),38.1％ (16/42) and 45.2％ (19/42),respectively.Multivariate analysis revealed that lesion range ＞ 3/4 esophageal circumference (odds ration [OR]:44.2 ; 95％ confidence interval [CI]:4.4-443.6) and tumor invasion beyond m2 (OR:14.2; 95 ％ CI:2.7-74.2) were independent risk factors.Stricture level was related to lesion's circumferential extension (relational coefficient (φ) =0.47,P ＜ 0.05) and tumor invasion depth (relational coefficient (φ) =0.647,P ＜ 0.05).Conclusion Circumferential extension and invasion depth of early esophageal cancer were independent risk factors for post-ESD esophageal stricture and related with the degree of stricture.

Based on the results of the Kirkwood high-dose interferon alpha-2b (HDI) adjuvant therapy trial of the Eastern Cooperative Oncology Group 1684, the US Food and Drug Administration (FDA) approved HDI as the postoperative adjuvant therapy for high-risk melanoma. Unfortunately, controversies continue regarding the use of interferon (IFN) as adjuvant therapy for melanoma owing to the inconsistent results of subsequent trials. Numerous trials of adjuvant interferon therapy demonstrated a benefit in terms of relapse-free survival (RFS), but without confirmed significant effect on overall survival (OS). The optimal timing, dose, and type of interferon are not yet defined. Therefore, adjuvant interferon treatment is preferentially applied in the randomized clinical trials in specialized centers. Decisions about the appropriateness of adjuvant interferon alfa-2b treatment for patients should be made on an individual basis, after discussion with the patient, including an explanation of the potential benefits and side effects of interferon therapy. Moreover, we also need to use available regimens reasonably, seek feasible and predictable prognostic factors to serve patients with individualized therapy.
Chemotherapy, Adjuvant ; Disease-Free Survival ; Dose-Response Relationship, Drug ; Humans ; Interferon-alpha ; administration & dosage ; therapeutic use ; Melanoma ; drug therapy ; Polyethylene Glycols ; therapeutic use ; Recombinant Proteins ; administration & dosage ; therapeutic use ; Skin Neoplasms ; drug therapy ; Survival Rate

Child ; Contracture ; congenital ; therapy ; Humans ; Male ; Neck Muscles ; pathology ; Torticollis ; congenital