ve To improve the method of determination of aluminium in water. Methods The method of aluminium chrome azurol S-cetylpyridinium bromide spectrophotometry was used. Results The improved determi-nation of aluminium revealed perfect linear relationship in determining the standard aluminium solution series at the range of 0. 0-6.0?g/25 ml aluminium. The regression equation was calculated as y = 1. 031x + 0. 008(r=0.9994) . The detection limit was 0.02 mg/L aluminium. The coefficient variation was below 4.90% . The recovery rate ranged from 93.3% to 100.0% . Conclusion The improved method was rapid and easy to operate, suitable for determination of Al3+ in a large number of water samples.

The effects of DHAA-urea,a novel dehydroabietylamine(DHAA) derivatives,on cell viability and glucose metabolism,in hypoxia and normoxia human hepatoma HepG2 cells were investigated.Hypoxia cells were achieved using DMEM containing high concentration of glucose without serum and pre-incubating of CoCl_2 (final concentration 150 μmol/L) for 24 h.The antiproliferation effect of DHAA-urea was measured by colorimetric MTT assay.The cellular ATP concentration,the lactate dehydrogenase(LDH) and glucose-6-phosphate dehydro genase (G6PD) activity were detected by their kits.It was shown that DHAA-urea markedly inhibited cell viability,cellular ATP level,LDH and G6PD activity in either aerobic or anaerobic circumstance in a dose-and time dependent manner.This suggested that DHAA-urea possibly inhibited HepG2 cells growth via the inhibition of glucolysis and glucolysis-dependent ATP depletion.DHAA-urea could be a promising candidate in the development of a novel class of agents used for human hepatocellular carcinoma.

Objective To validate genetic suppression of metastastic capability of highly metastastic melanoma cells by endostatin transfection.Method pcDNA3.1-Endo eukaryotic expression vector contained insulin signal peptide sequence was transfected into highly metastatic mice melanoma cell strain B 16.The expression of endostain was detected by RT-PCR and Western blot experiment,melanoma cells were determined with adhere experiment,in vitro invasion and migration experiment and pulmonary metastasis experiment on C57BL/6 mice.Result Endostatin can obviously inhibit the capability of adherence,in vitro invasion and migration and pulmonary metastasis of melanoma cells.Among them,adhere inhibition ratio was 67.3%,in vitro invasion inhibition ratio was 48.4%,cell migration inhibition ratiowas 52.1%and pulmonary metastasis inhibition ratio was 67.3%.Conclusion Endostatin transfection can obviously inhibit the highly metastatie capability of melanoma cells.

In order to provide anatomical basis for transoral approach (TOA) in dealing with the ventro lesions of craniocervical junction, and the design and application of artificial atlanto-odontoid joint, microsurgical dissecting was performed on 8 fresh craniocervical specimens layer by layer through transoropharyngeal approach. The stratification of posterior pharyngeal wall, course of vertebral artery, adjacent relationship of atlas and axis and correlative anatomical parameters of replacement of artificial atlanto-odontoid joint were observed. Besides, 32 sets of atlanto-axial joint in adults' fresh bony specimens were measured with a digital caliper and a goniometer, including the width of bony window of anterior arch of atlas, the width of bony window of axis vertebra, the distance between superior and inferior two atlas screw inserting points, the distance between two axis screw inserting points etc. It was found that the width of atlas and axis which could be exposed were 40.2+/-3.5 mm and 39.3+/-3.7 mm respectively. The width and height of posterior pharyngeal wall which could be exposed were 40.1+/-5.2 mm and 50.2+/-4.6 mm respectively. The distance between superior and inferior two atlas screw inserting points was 28.0+/-2.9 mm and 24.0+/-3.5 mm respectively, and the distance of bilateral axis screw inserting points was 18.0+/-1.2 mm. The operative exposure position through TOA ranged from inferior part of the clivus to the superior part of the C3 vertebral body. Posterior pharyngeal wall consisted of 5 layers and two interspaces: mucosa, submucosa, superficial muscular layer, anterior fascia of vertebrae, anterior muscular layer of vertebrae and posterior interspace of pharynx, anterior interspace of vertebrae. This study revealed that it had the advantages of short operative distance, good exposure and sufficient decompression in dealing with the ventro lesions from the upper cervical to the lower clivus through the TOA. The replacement of artificial atlanto-odontoid joint is suitable and feasible. The design of artificial atlanto-odontoid joint should be based on the above data.
Atlanto-Axial Joint/*anatomy & histology ; Atlanto-Axial Joint/*surgery ; Bone Plates ; Bone Screws ; Cadaver ; Cervical Vertebrae/*anatomy & histology ; Cervical Vertebrae/surgery ; Equipment Design ; Internal Fixators ; Joint Prosthesis ; Models, Anatomic ; Odontoid Process/*surgery ; Prosthesis Design

Objective To confirm whether or not let-7b and miR-199a were significantly associated with malignant melanoma growth and proliferation. Methods An over -expression plasmid and an inhibitor, which targeted on let-7b and miR-199a, was constructed. B16F10 cells were divided into seven groups: control group, let-7b plasmid group, miR-199a plasmid group, empty plasmid group, let-7b inhibitor group, miR-199a inhibitor group, inhibitor control group. Foreign gene was transfected into B16F10 cells, let-7b and miR-199a expression were validated from RNA level, protein level and cell level. Results The relative let-7b or miR-199a gene expression of the let-7b plasmid group (3.8776±0.1372)and miR-199a plasmid group (2.8660±0.2821)were significantly higher than control group (P＜0.05), the relative let-7b or miR-199a gene expression of the let-7b inhibitor group (0.2057±0.0263) and miR-199a inhibitor group(0.2656±0.0253) were significantly lower than control group(P＜0.05). The cyclinD1 expression of the let-7b plasmid group(2.023±0.315) and let-7b inhibitor group (1.857±0.377) were significantly higher than control group (0.997±0.041) (P＜0.05), whereas, the Met expression of themiR-199a plasmid group (5.19±0.309) and miR-199a inhibitor group (4.87±0.044) were significantly higher than control group (2.2±0.198) (P＜0.05). The let-7b plasmid group and miR-199a plasmid group B16F10 cell growth rate were slower than control group, especially on the third day after transfection, the growth rate gradually dropped to the lowest value (P＜0.05). In addition, the apoptosis rates of the let-7b plasmid group and miR-199a plasmid group reach to (11.8±1.19)% and (11.3±1.59)%,which were significantly higher than control group (P＜0.05). Conclusions let-7b and miR-199a may be a negative regulator on the B16F10 cell growth and proliferation.

Objective To apply Nutritional Risk Screening-2002(NRS-2002) to perform primary screening for nutritional risk in patients with esophageal cancer who undergo radiotherapy, and assess their nutritional status, and to investigate the value of NRS-2002 in such patients.Methods A total of 97 patients who were diagnosed with esophageal cancer and underwent radiotherapy in Zhejiang Cancer Hospital from January 2010 to April 2014 were analyzed retrospectively.The Kaplan-Meier method was applied to analyze the difference in survival, and the chi-square test and the Pearson correlation analysis were applied to analyze the correlation between NRS-2002 score and blood parameters.Results Of all patients, 26.8%had nutritional risk before radiotherapy, which gradually increased with the progress of radiotherapy.The 1-year overall survival rates of the patients with NRS-2002scores of ≤3 and ≥4 on admission were 91.1%and 61.9%, respectively (P=0.010).As for the patients with the highest NRS-2002 scores of ≤2 and ≥3 during treatment, the 1-year overall survival rates were 94.2% and 77.5%, respectively (P=0.012).As for the patients with the lowest NRS-2002 scores of ≤3 and ≥4 during treatment, the 1-year overall survival rates were 91.3% and 54.5%, respectively ( P=0.018).The NRS-2002 score was correlated with prealbumin on admission and at week 1 of radiotherapy (P=0.000 and 0.002), and the NRS-2002 score was correlated with albumin at week 3 of radiotherapy (P=0.036).The multivariate analysis showed that the TNM stage of esophageal cancer and the highest NRS-2002 score during treatment were the independent prognostic factors in esophageal cancer (P=0.001 and 0.005).Conclusions The patients with esophageal cancer undergoing radiotherapy have high nutritional risk, and NRS-2002 score is the independent prognostic factor in these patients and can be used as a tool for primary screening for nutritional risk.

In order to provide anatomical basis for transoral approach (TOA) in dealing with the ventro lesions of craniocervical junction, and the design and application of artificial atlanto-odontoid joint, microsurgical dissecting was performed on 8 fresh craniocervical specimens layer by layer through transoropharyngeal approach. The stratification of posterior pharyngeal wall, course of vertebral artery, adjacent relationship of atlas and axis and correlative anatomical parameters of replacement of artificial atlanto-odontoid joint were observed. Besides, 32 sets of atlanto-axial joint in adults' fresh bony specimens were measured with a digital caliper and a goniometer, including the width of bony window of anterior arch of atlas, the width of bony window of axis vertebra, the distance between superior and inferior two atlas screw inserting points, the distance between two axis screw inserting points etc. It was found that the width of atlas and axis which could be exposed were 40.2±3.5mm and 39.3±3.7mm respectively. The width and height of posterior pharyngeal wall which could be exposed were 40.1±5.2mm and 50.2±4.6mm respectively. The distance between superior and inferior two atlas screw inserting points was 28.0±2.9mm and 24.0±3.5mm respectively, and the distance of bilateral axis screw inserting points was 18.0±1.2mm. The operative exposure position through TOA ranged from inferior part of the clivus to the superior part of the C3 vertebral body. Posterior pharyngeal wall consisted of 5 layers and two interspaces: mucosa, submucosa, superficial muscular layer, anterior fascia of vertebrae, anterior muscular layer of vertebrae and posterior interspace of pharynx, anterior interspace of vertebrae. This study revealed that it had the advantages of short operative distance, good exposure and sufficient decompression in dealing with the ventro lesions from the upper cervical to the lower clivus through the TOA. The replacement of artificial atlanto-odontoid joint is suitable and feasible. The design of artificial atlanto-odontoid joint should be based on the above data.

BACKGROUND:Adjustable piezoelectric effect can promote tissue regeneration and repair.Piezoelectric materials are widely used in weight-bearing tissue engineering. OBJECTIVE:To prepare a piezoelectric film material that can promote bone regeneration,and to explore its structural characterization,electrical output performance,biocompatibility,and effect of electrical output on osteogenic differentiation of rabbit bone marrow mesenchymal stem cells. METHODS:Using poly-3-hydroxybutyrateco/4-hydroxybutyrate(P34HB)as raw material,barium calcium stannate titanate powder(Ba0.94Ca0.06Sn0.08Ti0.92O3,BCST)was added according to mass ratios of 0%,5%,10%,15%,and 20%.Dichloromethane was added to solve P34HB,and the thickness of 150-200 μm BCST/P34HB piezoelectric film was prepared by vacuum drying method.After polarization in the oil bath,the surface morphology,crystal phase composition,piezoelectric coefficient and open circuit voltage were tested.The effect of BCST/P34HB electrical output at 110 Hz and 0.25 N force on the proliferation and osteogenic differentiation of rabbit bone marrow mesenchymal stem cells was tested. RESULTS AND CONCLUSION:(1)Scanning electron microscopy,X-ray diffraction,water contact angle,piezoelectric coefficient and electrical output performance tests showed that when the mass ratio of BCST increased to 20%,the BCST/P34HB piezoelectric film had good piezoelectric properties(d33=5.9 pC/N)and electrical output performance(180 mV),which was closer to the suitable range of 500 mV for electrical stimulation.(2)Live and dead staining showed that on the first day of co-culture,15%group and 20%group showed less red fluorescence.On the 5th day of culture,the number of green fluorescence in each group was significantly higher than that on the first day,and the red fluorescence was not observed in the 10%,15%and 20%groups,and only a small amount of red fluorescence was observed in the 0%and 5%groups.(3)On the 1st,3rd and 5th days of co-culture with rabbit bone marrow mesenchymal stem cells,Almar blue staining exhibited that the number of cells in each group showed an increasing trend with the increase of time.On the 5th day of culture,the number of cells in the 20%group was significantly more than that in the 0%group(P<0.05).(4)On day 10 of osteogenic induction,alkaline phosphatase staining results showed that the positive rate of the 20%group was significantly higher than that of the 0%group(P=0.000 1).On day 21,alizarin red staining and quantitative analysis of calcium nodules showed a similar trend to alkaline phosphatase staining.Compared with the 0%group,the 15%group and 20%group showed significant differences(P<0.01,P<0.000 1).(5)The results showed that 20%BCST/P34HB films had good piezoelectric properties,electrical output properties,biocompatibility and the ability of promoting osteogenic differentiation of bone marrow mesenchymal stem cells.