[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.

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 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.

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.

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

The average life expectancy in Macau is ranking the second in the world, the consumption of fresh medicinal plants is a profound culture in Macau. The paper focus on the distribution of the antioxidant herbs, a comprehensive investigation and analysis the amount of the plant resources was carried out. The antioxidant activity of alcohol extracts was determined by using the DPPH method, and six kinds fresh herbs with high antioxidant free radical activity were screened out. Reference to adult daily dose of vitamin C, it is calculated that the daily dose amount of fresh herbs is less than 200 g. For the expected shortage of resources and the ecological status of Macau, we give some suggestions of herbal introduction in population ecology reconstruction.
Antioxidants ; pharmacology ; Ecosystem ; Free Radical Scavengers ; pharmacology ; Macau ; Plant Extracts ; pharmacology

Objective To analyze the clinicopathological characteristics of upper gastrointestinal submucosal tumors ( SMTs ) . Methods Clinicopathological data of 1 743 patients with 1 775 upper gastrointestinal SMTs in our department from January 2008 to December 2012 were retrospectively analyzed. Results The first finding was that in 702 esophagus cases,leiomyoma(92?59%,650/702) was the most common type of esophageal SMTs. Second, in 1 045 gastric cases, there were 405 lesions at gastric fundus, the most common type of SMTs were 249 ( 61?48%) GISTs and 144 ( 35?56%) leiomyoma. In 307 lesions located at body,the most common type of SMTs were 143( 46?58%) GISTs and 90( 29?32%) leiomyoma. In 191 lesions located at antrum, the most common type of SMTs were 83( 43?46%) heterotopia pancreas and 45(23?56%) hamartoma, followed by 28(14?66%) lipoma and 20(10?47%) GISTs. In 142 lesions located at cardia, the most common type of SMTs was 110 ( 77?46%) leiomyoma. Third, in 28 duodenum cases, there were 19 lesions at duodenal bulb, the most common type of SMTs was 10 heterotopia pancreas, 4 Brunner gland adenoma and 3 GISTs. In 9 lesions located at descending duodenum,the most common type of SMTs was 4 lipomyoma, followed by 2 ectopic pancreas,1 GISTs and 2 others. Conclusion Leiomyoma is the most common type of esophageal SMTs. In gastric fundus and body, the most common type of SMTs are GISTs and leiomyoma. In antrum, the most common type of SMTs are heterotopia pancreas and hamartoma, but in cardia, that is leiomyoma.In duodenal bulb, the most common type of SMTs are heterotopia pancreas, Brunner gland adenoma and GISTs,and in descending duodenum, is lipomyoma.

Objective To identify the activity of HCV IRES translation differences and identify the relationship between HCV IRES translation activity and ROS in different concentrations of ferric ammonium citrate ( FAC) in-duction.Methods 1 ) Expression plasmid pCI-Rluc-HCV IRES-Fluc was confirmed by endonuclease digestion as well as luciferase transient expression in Huh-7 cell;2) Controlled by dual-luciferase reporter assay, the differ-ent translation activity of HCV internal ribosomal entry site ( IRES ) was examined in a concentration of 50 μmol/L and 300μmol/L of FAC induction;ROS fluorescent staining method was used to detect the activity of ROS in Huh-7 cells, Western blot method was used to detect the protein expression changes of Nrf2 in Huh-7 cells;3) On the basis of the above experiments, 100 μmol/L DPI was added in 300 μmol/L FAC experimental group, to analyse the changes of HCV replication and ROS production after joining DPI.Results The generation of ROS and the activity of luciferase in the model group were significantly higher than that in the control group ( P<0.05 ) .FAC can enhance the expression of HCV IRES and increase the production of ROS , then causing Nrf2 expression in Huh-7 cell.However,after adding ROS inhibitor DPI, the above functions in Huh-7 cell were weakened.Conclusions The increase of HCV IRES expression induced by FAC is related to excessive ROS pro-duction induced by FAC in Huh-7 cells.