OBJECTIVE: To establish a method for the determination of the content of helicidum in helicid tablet intermediats.METHODS: The content of helicidum in intermediate was determined by the UV-visible spectrophotometry with the detective wavelength set at 270 nm.RESULTS: The linear range of helicidum was 4.05～12.15 ?g?mL-1(r=0.999 9) and its average recovery was 99.81%(RSD=0.35%).CONCLUSION: This method is simple,rapid and accurate,and suitable for the quality control of helicid tablet intermediats.

ObjectiveTo observe the effect of paroxetine on multiple sclerosis with depression and neurological functional rehabilitation.Methods60 patients with multiple sclerosis and depression were randomly assigned to two groups that were treated with paroxetine and fluoxtine. The effect was evaluated with Hamilton depression scale (HAMD), Hamilton anxiety scale (HAMA), activities of daily living scale (ADL) and P300 test.Results There were not differences in scores of HAMD, HAMA, ADL and P300 before and aftet the treatment in both paroxetine and fluoxtine groups (P>0.05), but there were significant differences in same group before and aftet the treatment (P<0.01).ConclusionParoxetine is effect on multiple sclerosis with depression effectively and able to accelerate neurological functional rehabilitation.

Objective To investigate the correlation between the variations of mitochondrial gene ATP6 and type 2 diabetes mellitus ( T2DM ) and chronic complications. Methods Genomic DNA were extracted from 254 T2DM patients and 165 age-matched controls. After amplification of ATP6 by PCR and direct sequencing, all sequences were compared with the reference sequence (rCRS) to find out the variations. Bioinformatics and statistic method were used to analyze these variations. Results Many variations were detected respectively in T2DM patients and controls, a part of them only appeared in T2DM patients in low frequency, which has not been reported previously. Most of these variations are located in thethird and forth transmembrane helix of ATP synthase subunit 6 (ATPase6). Interestingly, these variationsalmost were detected in the non-obese T2DM patients with hypotension, including G8557A, A8563G,T8594C, C8609T, A8689G, G8998A and G9139A. Conclusions There were many variations in geneATP6 and must of them are mitochondrial SNP, while variations A8689G, T8825C, G8920A, G8998A andG9139A may be mild mutations which my increase the susceptibility of T2DM. G8557A, A8563G,T8594C, C8609T, A8689G, G8998A and G9139A may be associated with the biogenetics diseases suchdiabetes and hypertension.

Objective To develop a rapid and sensitive method for the determination of bisphenol A in drinking water by liquid chromatography-electrospray tandem mass spectrometry. Methods Bisphenol A was extracted from the drinking water sample by a SEP-PAK C18 column,eluted with methanol and concentrated with a rotary evaporator. The extract was analyzed by liquid chromatography-tandem mass spectrometry with electrospray ionization. Results The calibration curve of bisphenol A was linear in the range of 5 -100 ng/ml,the linear equation was y =6 796.61x -8 655.64 and the correlation coefficients of linear calibration curve was 0.999 2. The limit of detection (S/N=3) and the limit of quantification (S/N=10) of bisphenol A were 0.007 5 ng/ml and 0.025 ng/ml,respectively. The rates of recovery of bisphenol A were from 83.46% to 94.00% and the relative standard deviation was 3.06% -4.60%. Conclusion The method is simple,rapid,accurate and is applicable to the qualitative and quantitative determination of bisphenol A in drinking water.

Objective To investigate the protective effects of thalidomide on rats with acute necrotizing pancreatitis (ANP) and its mechanism. Methods Fifty four SD rats were randomly divided into the three groups; ANP group, thalidomide group and control group with 18 rats in each group. The model of ANP was induced by retrograde injection of 5% sodium taurocholate into the bili-pancreatic duct. Rats in thalidomide group received thalidomide 200 mg/kg body weight gastric lavage 1 h after ANP induction. The rats were sacrificed 3 h, 6 h, and 12 h after ANP induction, and the amount of intraperitoneal ascites was quantified. The serum levels of TNF-α, IL-6, IL-18 were measured by EUSA. The proportion of CD4 + T cell, CD8 + T cell in peripheral blood was determined by flow cytometry. The expression of TNF-α mRNA in pancreatic tissue were measured by RT-PCR. The expression of ICAM-1 protein in pancreatic tissue was measured by immunohistochemistry. Pancreatic tissue underwent pathologic examination. Results Six hours after surgery, the amount of ascite, serum levels of TNF-α, IL-6, IL-18, CD4 + T cell, CD,+ T cell, pancreatic TNF-α mRNA and ICAM-1 protein expression, pathologic score in control group was (1.03 ±0.31)ml,(57.17±11.29)pg/ml, (24.45 ±4.14)pg/ml, (64.23 ±21.85)pg/ml, (47.58 ±9.21)% , (40.88 ± 2.96)%, 0.07 ±0.02, 0.57 ±0.30, 0.67 ±0.81, respectively, and the corresponding values were (3.63 ± 0.38)ml, (107.54 ±33.05) pg/ml, (47.30 ± 11.40) pg/ml, (367.76 ± 108.43 ) pg/ml, (54.90 ± 7.15)%, (17.17 ±3.12)%, 0.65 ±0.26, 3.20 ±0.57, 11.50 ±1.87 in ANP group; and (1.45 ±0.53)ml, (80.60 ±20.48) pg/ml, (26.61 ±10.85) pg/ml, (321.82 ±85.20) pg/ml, (29.80 ±2.19)% , (15.52± 1.96)%, 0.35 ±0.23, 2.37 ±0.67, 8.00 ±3.03. Besides the value of CD8 + T cell was significantly decreased, all other values were significantly increased when compared with control group (P <0.05). Conclusions Thalidomide can decrease the release of inflammatory mediator, and reduce the pathological damage of pancreas of ANP rats by inhibiting TNF-α mRNA expression.

Objective To investigate the therapeutic effects of melatonin (MT) on human pancreatic cancer in Balb/c nude mice subcutaneous xenograft model. Methods Pancreatic cancer model was established high dose MT(20 mg· kg -1 · d-1 ) , gemcitabine( GEM,50 mg· kg -1 · d-1 ) and high dose MT combination with GEM. Tumor size was measured regularly. The tumor growth curve was drawn. The activity of mice spleen natural killer cell was determined by MTT. Results Compared with the controls [ ( 1. 476 ± 0.075) cm3 ], tumor volumes in low dose MT group[ (0.998 ±0.112)cm3], high dose MT group[ (0.756 ±0.128) cm3], GEM group [ (0. 746 ± 0. 115 ) cm3 ], combination group [ (0. 305 ± 0. 111 ) cm3 ] were significantly decreased ( P <0.01), and the tumor size in high dose MT group was smaller than that in low dose MT group, while the tumor size in combination group was the smallest. The activity of mice spleen natural killer cell was ( 18.07 ± 1.23) %in control group, and they were (44.27 ±3.19)% ,(45.16 ±3.20)% and (30.29 ±2.91)% in low dose MT group, high dose MT group, combination group, which were significantly higher than those in the control group;the activity of natural killer cell in GEM group was ( 14.24 ± 2.70) %, which was significantly lower than that in the control group (P <0.05), but the activity of natural killer cell in combination group was significantly higher than that in the GEM group (P < 0.01 ). Conclusions MT could improve the immune function of mice,inhibit the growth of tumor, and MT combination with GEM may have more potent antitumor effect.

Objective To quantitatively detect the circulating DNA in plasma of patients with acute pancreatitis and to evaluate its potential clinical values. Methods Blood samples collected from 40 patients with mild acute pancreatitis (MAP), 20 with severe acute pancreatitis (SAP) and 50healthy controls were extracted for DNA using genomic DNA extraction kit. The DNA level was detected by real-time fluorescence quantitative PCR assay. The clinical association of DNA level with acute pancreatitis was analyzed. The receiver operating characteristic (ROC) curve was set up. Results The median concentration of plasma DNA in SAP group was 24.84 ng/ml, which was significantly higher than that in MAP group (7.60 ng/ml, P=0.006) and the healthy controls (5.23 ng/ml, P=0.000). The median concentration of plasma DNA in patients with acute pancreatitis was higher than in healthy controls (P=0.006), however, there was no significant difference between MAP group and healthy controls (P=0.322).The area under the ROC curve performed by the plasma DNA from SAP and MAP groups was 0. 881(95% CI, 0.773-0.989). With a cutoff value of 11.20 ng/ml, the overall sensitivity was 19/20% and specificity was 72.5%. Plasma DNA level was found to be associated with APACHE-Ⅱ score (P=0.001), Ranson score (P=0. 013), serum Ca2+ level (P=0.000), and c-reactive protein levels (P=0.001). Conclusions Plasma DNA is correlated with the extent of pancreatitis. It can be used in monitoring the development of acute pancreatitis and may be a potential marker for early diagnosis of SAP.

Objective:To investigate the feasibility,moderate-and short-term efficacies of laparoscopic surgery in the resection of the gastric gastrointestinal stromal tumors(GIST)with the tumor diameter more than 5 cm.Methods:The clinicopathological data of 26 gastric GIST patients with the diameter more than 5 cm underwent laparoscopic complete resection were retrospectively analyzed.The postoperative recovery,complications and moderate-and short-term prognosis of the patients were observed.Results:All the patients underwent laparoscopic surgery resection without open operation.According to the location and growth types of tumors,the total gastrectomy was performed in 1 case,the approximal gastrectomy was performed in 2 cases,the distal gastrectomy was performed in 2 cases,and the gastric local resection was performed in the rest 20 cases.The mean diameter of tumors was (5.94±1.28) cm,the mean operational time was (84.23±27.02) min,and the mean blood loss was (72.38±34.24) mL.The postoperative recovery time of bowel function was (2.77±0.65)d and the mean length of postoperative hospitalization time was (7.04±2.24) d.There was 1 case of intraperitoneal hemorrhage and 1 case of anastomotic stenosis;they recovered after conservative therapy.The postoperative pathology results(according to the modified NIH standard) indicated the number of moderate-risk patients was 18(69.23%),and the number of high-risk patients was 8(30.77%).A total of 12 cases accepted the adjuvant therapy with imatinib.There was 1 case of hepatic multimetastasis,and there was no local recurrence or death in the other patients.Conclusion:The moderate-and short-term efficacies of laparoscopic surgery in the gastric GIST patients with the tumor diameter more than 5 cm is safe and feasible.

ObjectiveTo study the change of cholinergic neurons of hippocampus of vascular dementia rats after transplantation of the neural stem cells(NSCs) in hippocampus.MethodsAfter the vascular dementia model was reproduced,the rats were randomly divided into transplantation,dementia and pseudo-operative groups.8 weeks after operation,immunofluorescence was used to observe the survivors and migration of neural stem cells.Immunochemistry staining was used to observe the number of ChAT positive neurons in hippocampus.ResultsThe number of the ChAT positive neurons in the CA1 subfield of the hippocampus were more significantly increased in transplantation group than that in dementia group. ConclusionChAT positive neurons can be found in hippocampus area of vascular dementia rats after transplanting NSCs into the hippocampus, which seem to be cholinergic neurons.

Objective To investigate the effect of pvdQ(PA2385)gene on Pseudomonas aeruginosa swarming motility.Methods The plasmid pME6032 with pvdQ gene was constructed and identified,then transformed into Pseudomonas aeruginosa PAO1 using the eleetroporation to build pvdQ-overexpression strain.The pME6032-PAO1 strain was constructed with the same method.The cloning plasmid pEX18Gm containing sacB was successfully used to construct unmarked deletion mutant of pvdQ gene and pvdQ mutant strain. Bacteria were inoculated in LB and were cultured overnight.The clones were measured for the diameter of the swarming zone.The statistical analysis was done using one-factor ANOVA.Results Strains of pvdQ-overexpression and pvdQ-mutant were successfully constructed and confirmed by polymerase chain reaction(PCR).The four strains were compared for the swarming motility by changes in diameter:PAO1(20.52±1.80)mm,pME6032-PAO1 strain(19.39±2.10)mm,pvdQ-overexpression strain(51.20±2.16)mm,pvdQ-mutant strain(3.30±0.55)mm.The diameter of pME6032-PAO1 strain was not significantly different from that of wild strain PAO1(t=-0.1493,P>0.05).However,the diameter of pvdQ(Q-mutant strain was significantly shorter than that of wild strain PAO1(t=2.8525,P<0.05).while the diameter of pvdQ-overexpression strain was longer than that of the wild strain PAO1(t=1.4230,P<0.05).Conclusions pvdQ gene may be involved in regulating the swarming motility of Pseudomonas aeruginosa,which can promote Pseudomonas aeruginosa swarming motility.