0.05), higher than those in the nighttime (t=3.63,3.16;P

Objective To establish a method used for determination of Aconitine and Hypaconitine and Mesaconitine in blood by LC/MS/MS.Methods Extraction of blood sample was conducted by use of 1% thirdchloroactic acid and acetonitrile liquid,and the electrospray fog ionic source,and positive ion MRM scan were employed.Result The linearity correlation coefficients(r)was≥0.992 2.The limit of quantification (LOQ S/N=5) was 2.0 ng/ml for Aconitine,0.5 ng/ml for Hypaconitine,and 0.5 ng/ml for Mesaconitine.The recovery of appended contrast in blank blood sample ranged from 91.25% to 103.1% with coefficient of variation (CV,n=6) less than 10.93%.Conclusion The method of LC/MS/MS is sensitive and reliable,and the sample handling is quick and simple,which was suitable for determination of the Aconitine and Hypaconitine and Mesaconitine in blood sample.

BACKGROUND:In the clinical treatment of diabetic retinopathy, the therapeutic effects of different filers on the retina and optic nerve after the removal of the vitreous are lack of comprehensive and detailed reports. OBJECTIVE:To observe the changes of retina and optic nerve in patients with diabetic retinopathy after filing cyclic polydimethylsiloxane. METHODS: Totaly 147 patients with diabetic retinopathy undergoing vitreous removal were enroled, including 77 males and 70 females, aged 35to70 years. The vitreous from 74 patients with diabetic retinopathy was filed with cyclic polydimethylsiloxane (test group). The cyclic polydimethylsiloxane was taken out at 3 months post-operation. The vitreous from another 73 patients with diabetic retinopathy was filed with the whole fluorine propane (control group). The changes of retina, optic nerve, intraocular pressure, and the occurrence of vitreous re-hemorrhage and cataract after operation were observed in these two groups. RESULTS AND CONCLUSION: After 7 days of folow-up, the intraocular pressure of patients in the test group was higher than that in the control group (P < 0.05). After 6 months of folow-up, there was no significant difference in the intraocular pressure between these two groups. After 6 months of folow-up, cyclic polydimethylsiloxane was successfuly removed in the test group, retinal blood vessels became thinning with the presence of white sheath, and the number of optic nerves was higher than the control group (P < 0.05). The incidence of vitreous re-hemorrhage and cataract in the test was higher than the control group (P < 0.05). These results demonstrate that after the removal of the vitreous, cyclic polydimethylsiloxane filing can lead to optic atrophy in patients with diabetic retinopathy.

RNA interference is a very important mechanism for the regulation of gene expression, participates in defense against viral infections, and keeps jumping genes under control. RNA interference has been taken as a powerful "gene silence" method widely used in basic science to study the function of genes, and it may lead to novel therapies in the future.

Objective To investigate the effects of IL-1 β on proliferation and migration of gallbladder cancer cells.Methods The secretion of IL-1 β in tissues of gallbladder cancer, chronic cholecystitis and normal gallbladder as well as in supernatant of gallbladder cancer cell lines (GBC-SD, SGC996) and HIBEpic cells was determined by enzyme-linked immunosorbent assay (ELISA) method.The levels of IL-1 β mRNA in GBC-SD, SGC996 and HIBEpic cells were measured by RT-PCR assay.The effects of exogenous IL-1 β on the proliferation of GBC-SD and SGC996 cells in vitro and in vivo were evaluated using WST-1 assay and xenograft tumor model, respectively.The effects of exogenous IL-1 β on the migration of GBC-SD and SGC996 cells in vitro were measured by Tranwell assay.The levels of Twist protein in GBC-SD and SGC996 ceils were examined by western blot assay after treatment with exogenous IL-1 β.In addition, the proliferation and migration of GBC-SD and SGC996 cells after gene silencing of Twist by Twist-siRNA were also evaluated.Results The level of IL-1β protein in normal gallbladder was low (66.4 ± 35.0)pg/ml,while it was significantly increased in gallbladder cancer and chronic cholecystitis [(616.4 ± 95.7) pg/ml and (422.3 ± 48.9) pg/ml, P ＜ 0.05].The levels of IL-1 βin GBC-SD and SGC996 cell culture medium [(587.4 ± 99.8) pg/ml and (657.2 ± 76.6) pg/ml] were much higher than those in the HIBEpic cells [(38.4± 12.1)pg/ml, P ＜ 0.05].Exogenous IL-1β promoted the proliferation of GBC-SD and SGC996 cells both in vitro and in vivo as well as migration in vitro (P ＜ 0.05).The level of Twist protein was significantly increased after treatment with exogenous IL-1 β.In addition, gene silencing of Twist blocked IL-1 β-induced proliferation and migration of GBC-SD and SGC996 cells.Conclusion IL-1 β promoted proliferation and migration of gallbladder cancer cells via Twist activation.

The crystal structural data of TACE, MMP-1, MMP-2, MMP-3 and MMP-9 were obtained from PDB database, and then their catalytic domains' properties including conformation, molecular surface hydrophobicity and electrostatic potential were analyzed and compared by using Insight II molecular modeling software. It was found that the conformation and molecular surface hydrophobicity of catalytic domains of TACE and MMPs were not obviously different, but the molecular surface electrostatic potential of catalytic domain of TACE and MMPs had obvious differences. The findings are helpful in the Rational Drug Design of TACE selective inhibitor.

Objective To explore the value of plasma C-peptide levels in early prediction of type 2 diabetes mellitus with peripheral sensory neuropathy.Methods The vibration perception threshold,pain,temperature sensation,touch-pressure sensation,ankle reflex was detected in 500 eases of type 2 diabetes mellitus,and the patients were divided into 4 groups according to peripheral sensory nerve test results:normal group (159 cases),mildly abnormal group (120 cases),moderately abnormal group (121 cases) and severely abnormal group (100 cases).Fasting and 2-hour postprandial C-peptide levels were determined and analysed with peripheral sensory nerve changes.The receiver-operating characteristic (ROC) curve was used to find the best critical point for diagnosis of diabetic peripheral sensory neuropathy.Results The fasting C-peptide among 4 groups had no significant difference (F =1.632,P ＞0.05).Two-hour postprandial C-peptide from normal group to mildly abnormal group and then moderately abnormal group gradually increased [(1.110 ± 0.526),(1.324 ± 0.490),(1.573 ± 0.716) μ g/L],while 2-hour postprandial C-peptide in severely abnormal group was significantly decreased and lower than that in normal group,and there were significant differences (P＜ 0.05).The max Youden Index was 0.366 when 2-hour postprandial C-peptide was 1.173 μ g/L.Conclusions The fasting C-peptide might be not related to early diabetic peripheral sensory neuropathy,but 2-hour postprandial C-peptide might be closely related to early diabetic peripheral sensory neuropathy.It is helpful to detect the early diabetic peripheral sensory neuropathy if we can take a dynamical observation of 2-hour postprandial C-peptide.

Essential hypertension (EH), a complex polygenic disease, is considered to the result of the genetic interaction of multiple gene alterations in concert with environmental factors. Evidences showed that angiotensin-converting enzyme (ACE) gene and G protein beta3 subunit (GNB3) gene are both important susceptibility genes for EH, and that there exists putative biological connection between the two genes in developing hypertension. To investigate whether hypertension was affected by gene-gene interaction between the two genes in the northern Chinese Han population, a case-control association study including 502 hypertensive cases and 490healthy controls was conducted, selecting the ACE gene I/D polymorpinsm and the GNB3 gene C825T polymorphism. Linkage disequilibrium analysis revealed a significant nonrandom distribution only in male hypertensives, indicating that interaction between ACE gene and GNB3 gene may predispose males to the occurrence of hypertension. Multivariate stepwise logistic regression in single locus analysis, with adjustment for common risk factors for hypertension, demonstrated that the OR for DD/ID versus Ⅱ for hypertension among men was significant (OR 1.57; 95% CI, 1.09 ～2.27; P = 0.016) in dominant genetic model. In combination analysis stratified with respect to gender, slightly significant ORs were found after adjustment in males: OR for TT vs CC, 0.11; 95%CI, 0.01 ～0.99; P = 0.049 within ACE DD genotype; OR for DD/ID vs Ⅱ, 1.52; 95% CI, 1.01 ～2.29; P = 0.047 within GNB3 CC+CT genotype. The results suggest that ACE, or a nearby gene, is a male-specific susceptible gene for hypertension, and that there may exist epistatic gene-gene interaction between ACE D allele and GNB3 825C allele.

Objective To establish the quality standard of Radix Toddaliae Asiaticae. Methods Thin layer chromatography ( TLC) and high performance liquid chromatography ( HPLC) were used to identify and determine chloride nitidine and toddalolactone in Radix Toddaliae Asiaticae. The moisture and total ash contents were detected according to the methods recorded in appendix of Chinese Pharmacopeia (2010 edition) . Results Toddalolactone and chloride nitidine were detectable by TLC, the spots were clear and the dissociation was good. The established HPLC method was simple and accurate. The linear ranges of toddalolactone and chloride nitidine in Radix Toddaliae Asiaticae were 2.84~42.6 μg/mL and 25.6~385 μg/mL, and their recovery rates were 99.2 % ( RSD=1.12%) and 100 % ( RSD=0.71%) , respectively. The content of moisture was in the range of 75.8~98.9 mg/g and that of total ash was in the range of 12.4~33.6 mg/g. Conclusion The developed method is specific and accurate, and can provide useful reference for establishing quality standard of Radix Toddaliae Asiaticae.

The crystal structural data of TACE, MMP-1, MMP-2, MMP-3 and MMP-9 were obtained from PDB database, and then their catalytic domains' properties including conformation, molecular surface hydrophobicity and electrostatic potential were analyzed and compared by using Insight Ⅱ molecular modeling software. It was found that the conformation and molecular surface hydrophobicity of catalytic domains of TACE and MMPs were not obviously different, but the molecular surface electrostatic potential of catalytic domain of TACE and MMPs had obvious differences.The findings are helpful in the Rational Drug Design of TACE selective inhibitor.