BACKGROUND: Studies on encapsulated whole cel protein antigen ofHelicobacter pylori are stil at the exploration stage. There is limited literature concerning the preparation process andin vitro release characteristics of chitosan microspheres encapsulated with whole cel protein antigen ofHelicobacter pylori. OBJECTIVE:To explore the preparation process andin vitrorelease characteristics of chitosan microspheres encapsulating whole cel protein antigen ofHelicobacter pylori. METHODS: Precipitation method was used to prepare chitosan microspheres, and the best preparation process, matching and encapsulation time were screened. Under electron microscope, the morphology and particle size of microspheres were observed. Chitosan microspheres were used to encapsulateHelicobacter pylori whole cel protein antigen, and BCA method was used to determine encapsulation efficiency, encapsulation content and release efficiency in vitro of Helicobacter pylori whole cel protein antigen. RESULTS AND CONCLUSION:Final concentration of 1% glacial acetic acid, sodium sulfate as crosslinking agent, pH=5.0, with no pulverization when the crosslinking agent was added was the best preparation process for chitosan microspheres. Electron microscopy showed the smooth surface morphology of microspheres with roundness and good dispersion, and the majority of the microspheres were 1-5 μm in diameter. The encapsulation efficiency ofHelicobacter pylori whole cel protein antigen microspheres was 80.4%, the encapsulated amount was 16.4%, and total 48-hour release rate was 19.4%.Helicobacter pylori whole cel protein antigen microspheres showed an overal slow release status. Chitosan microspheres show good encapsulation efficiency and amount ofHelicobacter pylori whole cel protein antigen, and Helicobacter pylori total bacteria protein antigen microspheres show an overal slow release status.

Objective To investigate the clinical significance of changes of serum amylase, CRP and SAA in the diagnosis of acute pancreatitis. Methods The levels of serum and urine amylase, CRP and SAA in patients of mild acute pancreatitis (MAP) and severe acute pancreatitis (SAP) at 24 h, 48 h, 72 h and the seventh day after the onset of pancreatitis were measured. Results The levels of serum, urine amylase, CRP and SAA in SAP patients at 24h were (904.5±402.2)U/L, (2280.3±1207.3)U/L, (155.6±36.2) mg/L, (521.9±109.4)mg/L, respectively, and significantly higher than those of MAP patients (P＜0.05 or P＜0.001). The peak value of serum amylase appeared at 24h, however, the peak value of urine amylase, CRP and SAA appeared at 48 h, and the corresponding values were (2173.5±1110.6) U/L, (185.3±41.4) mg/L and (717.5±144.2)mg/L, respectively. The levels of serum and urine amylase significantly decreases in MAP and SAP patients at the seventh day (P＜0.05). The levels of serum CRP and SAA significantly decreased in MAP patients at the seventh day (P＜0.05), however, the levels of serum CRP and SAA did not significantly decrease in SAP patients at the seventh day (P＞0.05). Serum levels of CRP and SAA were related to the severity of acute pancreatitis. Meanwhile CRP showed a positive correlation with SAA (r = 0.761, P＜0.05). Conclusions The change of serum levels of amylase, CRP and SAA can help early diagnose acute pancreatitis; CRP and SAA may predict the development of SAP at early stage.

Objective To optimize the matrix prescription and preparation technology of compound pyocutaneous gels. Methods Use the method of L9(34 ) orthogonal test, take the Carbopol-940 quantity, pH, glycerol and ethanol as factors,and take viscosity, pH value, appearance properties, temperature resistance as the comprehensive evaluation index, and compare the effects of stewing, centrifugal and ultrasound methods on removing bubbles in the gel. Ultimately determine the optimum preparation process. Results The best matrix prescription of compound pyocutaneous gels is as follows: carbopol-940 1.4%, 10% sodium hydroxide solution 3.3%,glycerol 2.3%,ethanol 6%;the best way to remove bubbles is centrifugation,with rotation rate at 3 000 r?min-1 for 10 min. Conclusion The selected matrix formulation is simple and feasible, the preparation technology is stable and reliable with good reproducibility, and can be used for the preparation of compound pyocutaneous gel.

Objective To explore the effect of support with total parenteral nutrition(TPN)or early enteral and parenteral nutrition(EN+PN)on immune function of critically ill neurosurgical patients.Methods In this prospective control study,patients were divided inte TPN group and EN+PN group based on the timing of admission.The changes of immunological indicators including CD3,CD4,CD8,CD4/CD8,CD3/CD25,IgA,IgG,IgM,and serum protein before and after nutritional support were compared.Results The percentage of T lymphocyte subsets CD3,CD4,and CD8,the ratio of CD3+/CD25+,the plasma leveh of IgA,IgM,and IgG,and the serum protein were significantly increased after nutrifional supports(P<0.05,P<0.01).However,compared with the TPN group,the percentages of T lymphocyte subsets(CD3,CD4,and CD8),the ratio of CD4+/CD8+,the plasma levels of IgA,IgM,and IgG,and the serum protein were significantly higher in EN+PN group(P<0.05,P<0.01).Conclusions Both TPN and EN+PN can promote the recovery of immune function,while EN+PN is superior to TPN.Early nutritional support should be provided to critically ill neurosurgical patients.

Objective To investigate the expression of RECK and MMP-9 in pancreatic cancer and to explore the relationship between RECK, MMP-9 expression and the clinicopathological characteristics.Methods PV6000 immunohistochemical method was used to detect the expression of RECK and MMP-9 in 28 cases of pancreatic cancer and 10 cases of normal pancreatic tissue. All the statistical analyses were performed by using SPSS 13.0 statistical software to determine the relationship between RECK, MMP-9 expression and the clinicopathological characteristics. Results The overall positive rate of RECK espression was 46.43% (13/28)in pancreatic cancer, which was significantly lower than that in normal pancreatic tissue (90%, 9/10). The positive rate of RECK espression in Ⅰ + Ⅱ clinical stage (75.0% ,9/12) was significantly higher than that in Ⅲ + Ⅳ stage (25.0%, 4/16 P < 0.05 ). The positive rate of RECK expression in cases without distant metastases (60.0%, 12/60) was significantly higher than that in cases with distant metastasis (12.5%, 1/8,P<0.05). The overall positive rate of MMP-9 was 75% (21/28) in pancreatic cancer, and 20% (2/10) in normal pancreatic tissue. The comparison between these two groups indicated a significant difference (P <0.01 ). The positive rate of MMP-9 in Ⅰ + Ⅱ clinical stage(50.0% ,6/12) was significantly lower than that in Ⅲ + Ⅳ stage (93.8,15/16, P < 0.05). The positive rate of MMP-9 in well differentiation group(33.3%,1/3 ) was significantly lower than that in poor differentiation group ( 100%, 12/12 ,P < 0. 01 ). The expressionof RECK was negatively correlated with the expression of MMP-9 ( r = - 0. 536, P < 0.01 ). Conclusions RECK is lowly expressed in pancreatic cancer, but MMP-9 is highly expressed. RECK and MMP-9 may serve as important markers in the evaluation of tumor stage.

Objective To evaluate the value of the Bedside Index for Severity in Acute Pancreatitis (BISAP) in diagnosing severe acute pancreatitis. Methods Sixty-eight patients with suspected diagnosis of severe acute pancreatitis were collected and were scored by BISAP, APACHE Ⅱ , Ranson and CTSI scoring systems, respectively. BISAP scoring system included the blood urea nitrogen, impaired mental status,systemic inflammatory response syndrome, age, and pleural effusion. The diagnosis criteria of severe acute pancreatitis was BISAP ≥ 3 points or APACHE IⅡ ≥ 8 points, Ranson ≥ 3 points, CTSI ≥ 3 points. The diagnostic accuracy of SAP of these scoring systems was calculated. Results Among these 68 cases, 63.2％(43/68) were graded ≥ 3 points in BISAP scoring system;60.3％ (41/68) were marked ≥8 points in APACHE Ⅱ scoring system; 60.3％ (41/68) were scored ≥ 3 points in Ranson scoring system; and 67.6％(46/68) were scored ≥3 points in CTSI scoring system. There was no statistical difference between BISAP scoring system and other three scoring systems in diagnosing severe acute pancreatitis. Conclusions As a new and simple scoring system, BISAP scoring system can be widely used in the diagnosis of severe acute pancreatitis.

Objective To detecte the expression of COX-2,VEGF-C and lymphatic vessel density (LVD)in pancreatic cancerous and paracancerous tissues,and investigate their correlation.Methods The expression of COX-2.VEGF-C and LVD in 40 cases of pancreatic cancer tissues and paracancerous tissues and 12 cases of normal pancreas was detected by tissue chip and immunohistochemical assays,and the relationship between them and the cljnicopathological parameters was analyzed. Results The expression of COX-2,VEGF-C in pancreatic cancer tissues were 70.0%(28/40)and 67.5%(27/40),respectively,which were significantly higher than that in paracancerous tissues(42.5%,17/40)and(35.0%,14/40),and that in normal pancreas(8.3%,1/12)and(25.0%,3/12).The LVD in pancreatic cancerous,paracancerous and normal pancreatic tissues were 4.75±2.77,15.2 ±4.70 and 1.67±1.15,respectively.The expression of COX-2 in cancerous tissues and LVD in paracancerous tissues was correlated with tumor differentiation and lymph metastasis;the expression of VEGF-C Was correlated with lymph metastasis.LVD in paracancerous tissues was correlated with the expression of COX-2 and VEGF-C.Conclusions Pancreatic cancer lymphangiogenesis mainly existed in paracancerous tissues,COX-2 and VEGF-C may play an important role in the lymphangiogenesis.

Proteomic analysis of core needle biopsy (CNB) sample from patient populations is critical to our understanding of human disease,but has been hindered by its particular small size.Here,we present a method for the proteomic analysis of CNB sample based on the two dimensional electrophoresis.Proteins were extracted directly from 3 rat liver CNB specimens and a human prostate CNB sample.respectively.24 cm Immobiline DryStrip (pH 3-10NL) and 12.5% SDS-PAGE were introduced to separate the proteins.Interesting spots were analyzed by MALDI TOF/TOF mass spectrometry after tryptic digestion.With this method,consistent electrophoretic patterns of more than 2 500 protein spots were reproducibly obtained after silver staining,from rat liver CNB specimens.Qualitatively and quantitatively reproducible results also yield when the method was applied to a human prostate CNB sample.57 stochastically selected protein spots were analyzed by MALDI TOF/TOF moss spectrometry.and were identified with high confidence including faint ones.This simple and reproducible approach raises the opportunity of defining key molecular events of human disease pathologies.

Objective To investigate the apoptosis-inducing effect and anti-proliferative effect of epigallocatechin-3-gallate (EGCG) on human pancreatic cancer cell SW1990 in vitro. Methods The effect of proliferation was evaluated by MTT after the SW1990 cells in vitro were incubated with different concentrations of EGCG (6.25, 12.5, 25, 50, 100 μg/ml). The apoptosis-inducing effect was determined by flow cytometry after the cells were treated with 25 μg/ml of EGCG. The cell cycle of SW1990 cells was detected by flow cytometry after the cells incubated with different concentrations of EGCG (0, 10, 20, 30, 40, 50 μg/ml).Results After SW1990 cell were treated with different concentrations of EGCG (0, 25, 50 μg/ml), the values of A492 were 0.46 ±0.04,0.42 ±0.04,0.27 ±0.03 at 24 h; 0.48 ±0.02, 0.31 ±0.03,0.16 ±0.02at 48 h; 0.51 ±0.01,0.24 ±0.04,0. 14 ±0.04 at 72 h. EGCG inhibited the proliferation of SW1990 in a doseand time-dependant manner(P ＜0.01 ). The apoptotic rates at 24, 48, 72 h were (8.33 ± 1.15 )%, (19.77 ±0.81 )%, (29.17 ± 0.75 )% in the EGCG treatment group; while the corresponding values were (2.77 ±0.45 ) %, (3.20 ± 0.26 ) %, (3.67 ± 0.35 ) % in the control group; and the difference was statistically significant (P ＜0.01 ). After 0, 20, 50 μg/ml of EGCG treatment for 24 h, the percentages of SW1990 cellsin G0/G1 stage were (57.59 ±0.97)%, (62.99 ± 1.91 )%, (68.87 ± 1.88)%, and the percentages of SW1990 cells in G0/G1 stage increased with the increase of concentrations of EGCG, while the percentages of SW1990 cells in G2/M stage decreased with the increase of concentrations of EGCG (P ＜0.01 ). Conclusions EGCG can significantly inhibit the proliferation of SW1990 cells. The mechanism may be related to the apoptosis-inducing effect and the regulation of the cell cycle of the SW1990 cells.

Primary liver cancer is a common malignant tumor with complex pathogeneses and has the features of insidious onset, high degree of malignancy, and poor prognosis. The change in glycolytic pathway is one of the most important differences between tumor cells and normal cells, and tumor cells prefer to generate energy from glycolysis. Aerobic glycolysis is often associated with the progression and poor prognosis of primary liver cancer. Long non-coding RNAs (lncRNAs) can influence the glycolysis pathway in many tumors by regulating glucose uptake and the expression and activation of glycolytic enzymes and thus play an important role in the development and progression of primary liver cancer, which suggests that lncRNAs can be used as a therapeutic target for liver cancer. This article summarizes the influence of lncRNAs on primary liver cancer and glucose metabolism and related mechanisms, so as to find potential and effective targeted therapies for primary liver cancer.