OBJECTIVE:To optimize the formula technology of puerarin derivative 4ac-loaded nanoparticles by central composite design. METHODS:4ac nanoparticles were prepared using solvent evaporation method, with mean diameter, encapsulation rate and yield as dependent variables, multi-linear regression and binomial fit were performed on the 3 independent variables at different level:dosages of supporter lactic acid/ glycolic acid copolymer and surfactant polyvinyl alcohol, and the diameter of pinhole. The optimum experimental conditions were predicted by response surface method. RESULTS:The binomial fitting equations of indexes were all superior to their multi-linear regression equations, and the predictive values in the established mathematical model were in good conformity with the experimental values. CONCLUSION:Central composite design plus response surface method shows a good predictability in optimizing the formula technology of 4ac-loaded nanoparticles.

Objective To explore the application effect of hyperbaric oxygen ( HBO ) in the treatment of refractory peptic ulcer .Methods Clinical data of 259 patients with refractory peptic ulcer were retrospectively ana-lyzed.According to whether received HBO therapy , these patients were divided into the HBO group ( n=127 ) and control group(n=132).The total effective rate,Helicobacter pylori(Hp) eradication rate and relapse rate of the two groups were compared .Results The total effective rate ,Hp eradication rate of the HBO group were 93.7%,92.1%, respectively,which were significantly higher than 65.2%,56.1% of the control group (χ2 =31.97,7.32,all P<0.05).The relapse rate of the HBO group was 8.4%,which was significantly lower than 33.3%of the control group (χ2 =12.51,P<0.05).Conclusion HBO combined with conventional drug in the treatment of refractory peptic ulcer has significant effect .

AIM:To establish a method for determining the contents of salvianolic acid B,danshensu and protocatechuic aldehyde,the effective components of Shuangdan Oral Solution(Radix et Rhizoma Salviae miltiorrhizae,Cortex moutan) by HPLC.METHODS:Experiment was carried out on a Phenomenex Luna C_(18) column(4.6 mm?150 mm,5 ?m) with a mobile phase methanol-acetonitrile-methane acid-water(30:10:1:59) at a flow velocity of 1.0 mL/min and detection wavelength at 286 nm for salvianolic acid B,a mobile phase methanol-acetonitrile-methane acid water solution [HCOOH:H_2O(2:98)](9:4:87) at a flow velocity of 1.0 mL/min and detection wavelength at 281 nm for danshensu and protocatechuic aldehyde.RESULTS:The average recoveries of salviano-(lic) acid B,danshensu and protocatechuic aldehyde were 98.6%(RSD=1.91%,n=6),99.2%(RSD=2.2%,n=6) and 97.3%(RSD=2.5%,n=6) respectively.CONCLUSION:The results show that this method is easy to operate and accurate,and can be used to determine the contents of the three effective components in Shuangdan Oral Solution.

Objective To investigate the correlation between plasma D-dimer with delayed encephalopathy (DE) in patients with acute carbon monoxide poisioning(ACOP).Methods According to the severity of disease,100 patients with ACOP were divided into mild group (27 cases),moderate group (31 cases),severe group (42cases).And depending on whether DE occured,all patients were divided into DE group(20 cases) and no DE group (80 cases).4 venous blood samples were obtained separately from each subject in 1,7,14 days after their admission to hospital by using latex immunoturbidimetry.The levels of blood plasma D-dimer of patients in each group were detected by immune turbidimetry and analyzed,1th,7th and 14th day after admission.Results The plasma D-dimer level of the mild group at different time points was in the normal range,and the difference was not statistically significant(all P ＞ 0.05) ;The plasma D-dimer level of the severe group and moderate group was significantly higher than that of the mild group(all P ＜0.01),and it reached a peak in one week(7 days),and showed a downward trend after two weeks(14 days).The DE all occurred in the moderate and severe group.The plasma D-dimer levels of the DE group were higher than those of no DE group from admission to the 7 th and 14 th day (all P ＜ 0.01).Conclusion The dynamic monitoring plasma D-dimer is helpful in assessing to the severity and prognosis of ACOP,and could surpport positive clinical significance for the prevention and treatment of DE.

Objective To assess the effect of three different liver vascular exclusions on prognosis of patients undergoing hemihepatectomy.Methods Clinical data of 216 patients undergoing hemihepatectomy were analyzed retrospectively.Ninety-eight out of 216 patients received Pringle maneuver during hepatectomy in group A,71 patients of selective liver inflow and outflow vascular exclusions in group B,47 patients using liver hanging maneuver combining with selective liver inflow and outflow vascular exclusions were in group C.Results There was no difference in operation time between the three groups ( t =0.72,0.83,and 0.67,P ＞ 0.05 ).The intraoperative blood loss and transfusion in group B and C were less than that in group A (t =3.72,3.83 and 4.11,4.07,P ＜0.05).Serum albumin level on day 1 and day 3 in group B and C were higher than that in group A (t =3.65,3.77,and 3.90,3.74,P ＜0.05 ).Serum total bilirubin level on day 3 and 5 in group B and C were lower than that in group A ( t =4.13,5.01,and 4.09,3.99,P ＜0.05).Serum alanine aminotransferase on day 1,3 and 5 in group B and C were lower than that in group A ( t =5.36,6.14,and 5.70,7.01,and 4.94,3.98,P ＜ 0.05 ).Postoperative complication rate in group A was higher than that in group B and C ( x2 =13.71 and 23.56,P ＜ 0.05 ).The 3-year survival rate of patients with malignant tumor in the three groups were not significantly different (t =2.38,P ＞ 0.05 ).Conclusions Intraoperative blood loss and transfusion and postoperative complication rate can be reduced,and liver injury can be diminished in hemihepatectomy using selective liver inflow and outflow vascular exclusion alone and or in combination with a liver hanging maneuver.

Objective To explore the expression and clinical significance of Twist. E-cadherin and N-cadherin in gallbladder carcinoma. Methods From 2000 to 2008, in Zhejiang Provincial People's Hospital, the expression of Twist, E-cadherin and N-cadherin protein were detected in 79 surgically removed gallbladder carcinoma tissue specimens in paraffin blocks and 20 normal gallbladder tissue specimens by tissue microarray technique and immunohistochemistry. Results Compared with normal gallbladder tissues, the expression of Twist and N-cadherin was up-regulated in gallbladder carcinoma tissues and the positive percentage was 68.3％ and 49.4％ respectively, while which both were 1/20 in normal gallbladder tissues. The percentage of high E-cadherin expression in gallbladder carcinoma tissues was only 27.8 ％, which was significantly lower than that in normal gallbladder tissues (20/20;X2 ＝29.31, P＜0. 05). The expression of Twist was correlated with T classification, lymph node metastasis, distant organ metastasis, hepatoduodenal ligament invasion, lymphatic invasion and UICC stage of gallbladder carcinoma (P＜0.05). The expression of E-cadherin was correlated with T classification, distant organ metastasis, hepatoduodenal ligament invasion, differentiation degree and UICC stage of gallbladder carcinoma (P＜0.05). The expression of N-cadherin was only correlated with lymphatic invasion of gallbladder carcinoma (P ＜0.05 ). There was significant negative correlation between Twist and E-cadherin expression (P＜0. 01). All the 79 gallbladder carcinoma patients were followed up after the surgery, the mean follow-up time was 30.6±14.3months. The 3-year survival rates of patients with low or high Twist expressions were 66％ and 7％ respectively,there was significant difference between the two groups(P＜0. 01). The 3-year survival rates of patients with low or high E-cadherin expressions were 25 ％ and 86 ％ respectively, the difference of two groups was significant(P＜0.01). The 3-year survival rates of patients with low or high Ncadherin expressions were 39％ and 41％ respectively, there was no significant difference between the two groups(P＞0.05). The multivariate analysis indicated that the Twist expression was one of independent prognostic factors of gallbladder carcinoma. Conclusion The abnormal expression of Twist and E-cadherin was correlated with the development and progression of gallbladder carcinoma,and Twist expression was one of the independent prognostic factors of gallbladder carcinoma.

Objective To investigate the clinical influence about levetiracetam on cognitive function and emotional influence in the patients with partial epilepsy.Methods A total of 62 patients with partial epilepsy were divided into carbamazepine group (30 cases) and levetiracetam group (32 cases) by random digits table method,carbamazepine group was treated by carbamazepine while levetiracetam group was treated by levetiracetam.The patients were assessed before treatment and 4,8,12,16 weeks after treatment by the Montreal cognitive assessment scale (MoCA),self rating anxiety scale (SAS) and self rating depression scale (SDS).Results There was no significant difference in MoCA score between two groups before treatment and 4 weeks after treatment (P ＞ 0.05).MoCA score at 8,12,16 weeks after treatment in levetiracetam group was better than that in carbamazepine group [(22.6 ± 2.1) scores vs.(20.8 ± 2.6) scores,(23.5±2.7) scoresvs.(21.3± 2.8) scores,(24.6±4.7) scoresvs.(21.2±3.0) scores],the difference was statistically significant (P ＜ 0.05).There was no significant difference between SAS score,SDS score between two groups before and after treatment (P ＞ 0.05).Conclusion In the process of treating the patients with partial epilepsy by drug,levetiracetam is superior to carbamazepine on the improvement of cognitive function,but the mood improvement is not obvious.

Objective To explore the expression of Twist protein in the colorectal cancer (CRC) and its relationship between clinicopathological factors and prognosis of patients.Methods The clinical data of 45 patients with colorectal cancer who were admitted to the First People's Hospital of Shunde from June 2005 to September 2005 were retrospectively analyzed.The colorectal tissue specimens from surgical resection were collected.The expressions of Twist protein in the cancer and paracarcinoma tissues were examined by the immunohistochemistry,and their relationship with clinicopathological factors and prognosis of patients was analyzed.Patients were followed up by outpatient examination and telephone interview until September 2013.The count data were analyzed using the chi-square test and Fisher exact probability.The survival curve was generated using the Kaplan-Meier method,and the survival analysis was conducted using the Log-rank test.The prognosis factors were analyzed by the COX univariate analysis and multivariate analysis.Results The expression of Twist protein in CRC was predominantly detected in the cytoplasm of the tumor cells.The positive expression rates of Twist protein in the CRC and paracarcinoma tissues were 62.2％ (28/45) and 20.0％ (9/45),respectively,with a significant difference (x2 =18.383,P ＜0.05).The positive expression rates of Twist protein were 42.1％ in patients with Ⅰ-Ⅱ stage of CRC and 76.9％ in patients with Ⅲ-Ⅳ stage of CRC.The positive expression rates of Twist protein were 41.7％in patients with N0 stage of CRC and 85.7％ in patients with N1-N2 stage of CRC.The expression of Twist protein in patients with CRC was however associated with the TNM stage and lymph node metastasis (x2=5.662,9.244,P ＜ 0.05).A total of 45 patients were followed up for 10-96 months (median,54 months).The median survival time and 5-year cumulative survival rate in 45 patients with positive expression of Twist protein were 44 months and 10.7％,which were compared with 69 months and 82.4％ in 45 patients with negative expression of Twist protein,with a statical significance in the survival (x2 =26.147,P ＜ 0.05).The results of univariate analysis showed that the expression of Twist protein was an related factor affecting the prognosis of patients [HR =8.669,95％ confidence interval (CI):2.959-25.390,P ＜0.05].The results of multivariate analysis showed that the positive expression of Twist protein was an independent risk factor affecting the prognosis of patients (HR =5.059,95 ％ CI:2.888-8.863,P ＜ 0.05).Conclusiors The increasing expression of Twist protein in the CRC is positively correlated with the occurrence and development of CRC.The positive expression of Twist protein is an independent risk factor affecting the prognosis of patients with CRC.

Objective To observe the effects of total flavone from litchi chinensis sonn (TFL) on the liver function in-cluding p16 protein, pro collagen type 3 (PC3) and pro collagen typeⅠ(PCⅠ) in model rats with liver fibrosis induced by bile duct ligation. Methods Forty rats were randomly divided into four groups:sham operation (SO) group, bile duct liga-tion (BDL) group, TFL group and silibinin (SIL) group. Rats were gavaged with saline (5 mL·kg-1·d-1) in SO and BDL group, rats were gavaged with TFL (200 mL·kg-1·d-1) in TFL group and rats were gavaged with SIL (5 mL·kg-1·d-1) in SIL group for four weeks. The serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), bilirubin direct (BILD) and bilirubin total (BILT) were detected in four groups. The liver tissues were stained by HE and Masson methods. The ex-pression levels of p16, PC3 and PCⅠin liver tissues were determined by Western blot assay. Results The serum levels of ALT (44.6 IU/L±8.0 IU/L), AST (103.8 IU/L±18.1 IU/L), BILD (0.76 μmol/L±0.28μmol/L) and BILT (1.48μmol/L±0.35μmol/L) were lower in SO group. There was a higher level of ALT in BDL group (147.4 IU/L±86.3 IU/L) than that of TFL group (92.9 IU/L±47.3 IU/L). The serum level of ALT was higher in AST group (362.7 IU/L±106.6 IU/L) than that of TFL group (290.1 IU/L ± 171.7 IU/L) and SIL group (250.2 IU/L ± 54.9 IU/L). The serum level of BILD was lower in BDL group (99.71μmol/L±40.87μmol/L) than that of SIL group (137.01μmol/L±38.86μmol/L). The serum levels of BILD and BILT were significantly lower in TFL group (81.48μmol/L±47.50μmol/L, 106.64μmol/L±61.04μmol/L) than those of SIL group (P<0.05). There were small amount of new bile duct and no obvious cells degeneration, small amount of infiltration of in-flammatory cells and collagen deposition in TFL group. The liver fibrosis improved significantly in TFL group than that of BDL group. There were more new bile duct in hepatic portal area in SIL group than those of TFL group. The expression levels of p16, PC3 and PCⅠwere significantly higher in BDL group than those of TFL group. The expression level of PC3 was significantly lower in BDL group than that of SIL group. The expression level of PCⅠwas significantly higher in BDL group than that of SIL group (P<0.05). There was no significant difference in the expression level of p16 between BDL group and SIL group. The expression levels of PC16 and PC3 were significantly lower in TFL group than those of SIL group (P<0.05). There was no significant difference in the ex-pression level of PCⅠbetween TFL group and SIL group. Conclusion TFL can improve the liver function in model rats with choles-tatic liver fibrosis and reduce liver fibrosis, which may be related with inhibitory effects on the expressions of p 16, PC3 and PCⅠ.

Objective To study the causes of and management for reoperative surgery in patients of congenital choledochal cyst. Methods The clinical data of 41 patients of congenital choledochal cysts undergoing reoperative surgeries were reviewed. Results There were 32 cases of type Ⅰ , 1 of type Ⅱ ,3 of type Ⅳ, and 5 of type Ⅴ according to Todani classification of congenital choledochal cyst. Thirty patients experienced one previous biliary tract surgery, 8 had a history of 2 surgeries and 3 experienced three times of operations previously. Among the 41 patients, 40 presented bile duct stones, 33 suffered from biliary infections, 7 had hilar hepatic duct strictures, 6 had strictures of previous hepatoenteric anastomosis, 5 had carcinomas arising in cysts. The reoperative surgeries for the 41 patients consisted of excisions of extre-hepatic bile cyst and hepato-jejunal anastomosis by Roux-Y fashion (combining with hepatoduodenal ligament dissection and regional lymphadenectomy for two patients with cholangiocarcinoma) in 32 cases, hepatectomies combined with excisions of choledochal cyst and hepato-jejunal anastomosis in 8 cases, reconstruction of hepatoenteric anastomosis by excision of stenosis in one. All patients were followed up with mean time of 53. 7 ± 32. 1 months, ranging from 3 months to 10 years. Intermittent biliary infection was experienced in 4 cases during follow-up period, and there was no anastomotic stricture and stone recurrence. Conclusion The main causes of reoperation for congenital choledochal cyst patients with a history of previous surgery are complications arising from incorrect surgical procedure and misdiagnosis. Complete excision of extra hepatic bile cyst combined with Roux-en-Y hepatojejunal anastomosis benefits patients who had a failed previous surgery.