Aim To develop an HPLC method for the determination of Aconitum alkaloids extracted from Radix aconiti preparata in rats. Methods Waters 2690@996 PAD system was used. The analytical column was a Halsil 100 C18 column (250 mm×4.6 mm ID, 5 μm). The mobile phase was water, methanol and diethyl amine at the ratio of 75∶ 25∶ 0.1. The flow rate was 0.9 mL·min -1. The wavelength of the detector was 240 nm. Results The linear ranges of aconitine in the heart, spleen, lung and kidney were 0.4-100 μg·mL -1, the correlation coefficients were 0.997 2, 0.998 6, 0.999 3 and 0.999 4, respectively. The linear range of aconitine in liver was 2-200 μg·mL -1 and the correlation coefficient was 0.999 0. The linear ranges of hypaconitine in heart, liver, spleen, lung, kidney, brain and spinal cord were 5-100 μg·mL -1, the correlation coefficients were 0.999 4, 0.999 7, 0.999 8, 0.998 4, 0.999 8, 0.999 8 and 0.999 7, respectively. Detection limits (S/N=3) of aconitine and hypaconitine were 0.4 μg·mL -1. The recoveries of aconitine and hypaconitine ranged from 88.7% to 102.2% and 86.5% to 101.3%, respectively, and the RSD of precision of aconitine and hypaconitine was 10%. Conclusion It appears to be an accurate and effective method that can offer reference basis for in toxication of Radix aconiti preparata clinically.

Cone-Beam Computed Tomography ; Humans

Heart Defects, Congenital ; diagnostic imaging ; Humans ; Organ Size ; Radiography, Thoracic ; Syndrome

Objective To explore the different expression and clinical significance of miR-146a/133b in cervical tissue in uy-ghur and Han women in Xinjiang.Methods The relative expression of miR-146a/133b in paraffin embedding tissues of cervicitis, CIN and cervical cancer was detected by the RT-qPCR.And analyzed the clinical significance in the development of cervical cancer. Results Compared with cervicitis,the expression of miR-146a/133b increased significantly in CIN and cervical cancer(P <0.05). With the cervical lesion was aggravating,the expression level increased.In cervical cancer tissue,the expression of miR-146a were different between Uyghur and Han women(P <0.05).Marriage age<20 years old,tumor diameter≥4 cm,with HPV infection in cervical cancer tissue,miR-146a/133b had high expression (P <0.05).Conclusion MiR-146a/133b are involved in incidence and development of cervical cancer,they may become new prognostic and evaluating molecular markers in cervical cancer.

Objective To determine the effect of unfractionated heparin (UFH) on lipopolysaccharide (LPS)-induced expression of chemokines and nuclear factor-κB (NF-κB) signaling pathway. Methods Human pulmonary microvascular endothelial cells (HPMECs) were cultured in vitro, and the cells between passages 3 and 5 were used in the experiments. The cells were divided into control group, LPS challenge group, 1 kU/L or 10 kU/L UFH+LPS group, and NF-κB inhibitor N-tosyl-L-lysyl chloromethyl-ketone (TLCK) group (TLCK+LPS group). HPMECs in LPS challenge group were treated with 10 mg/L LPS. UFH pretreatment with different dosages groups were treated with 1 kU/L or 10 kU/L UFH 15 minutes before LPS challenge. Cells in the TLCK+LPS group were treated with 10 μmol/L of TLCK 30 minutes before the addition of LPS, and HPMECs in control group were treated with an equal volume of phosphate-buffered saline (PBS) instead. The cells were harvested 1 hour after LPS challenge, and the nuclear translocation of NF-κB was determined by immunofluorescence assay to detect the effect of UFH on NF-κB activation. The levels of interleukin-8 (IL-8) and monocyte chemoattractant protein-1 (MCP-1) in cell culture supernatants were determined by enzyme linked immunosorbent assay (ELISA) 3 hours and 6 hours after LPS challenge to detect the effect of UFH on LPS induced expression of chemokines and its mechanism of effect on NF-κB signaling pathway in HPMECs. Results ① In the control group, NF-κB was mostly located in the cytosol as shown by immunofluorescence. Treatment of HPMECs with LPS significantly increased the translocation of NF-κB from the cytosol to nucleus. UFH suppressed LPS-induced NF-κB activation both in 1 kU/L and 10 kU/L dosages, and 10 kU/L UFH gave even better results. ② Compared with control group, the levels of IL-8 and MCP-1 in the supernatants in LPS challenge group were significantly increased at 3 hours and 6 hours after LPS challenge [IL-8 (ng/L): 387.1±26.4 vs. 23.8±8.1 at 3 hours, 645.5±69.6 vs. 125.7±18.7 at 6 hours; MCP-1 (ng/L): 3 654.9±467.9 vs. 721.6±61.3 at 3 hours, 8 178.5±792.6 vs. 1 324.7±148.7 at 6 hours, all P < 0.05]. Compared with that of LPS challenge group, in 1 kU/L and 10 kU/L UFH pretreatment groups, the levels of IL-8 and MCP-1 were significantly decreased [IL-8 (ng/L): 315.3±24.8, 275.8±31.1 vs. 387.1±26.4 at 3 hours, 557.8±43.3, 496.9±38.7 vs. 645.5±69.6 at 6 hours; MCP-1 (ng/L): 2 924.1±267.9, 2 668.3±522.6 vs. 3 654.9±467.9 at 3 hours, 7 121.7±557.2, 6 563.9±576.4 vs. 8 178.5±792.6 at 6 hours, all P < 0.05]. The results indicated that 10 kU/L UFH yielded better results. However, inhibition study using the known NF-κB inhibitor TLCK could decrease LPS-induced increase in IL-8 and MCP-1 levels [IL-8 (ng/L): 162.4±21.3 vs. 387.1±26.4 at 3 hours, 274.1±22.6 vs. 645.5±69.6 at 6 hours; MCP-1 (ng/L): 1 478.2±138.5 vs. 3 654.9±467.9 at 3 hours; 3 667.6±259.4 vs. 8 178.5±792.6 at 6 hours, all P < 0.05]. Conclusions The levels of IL-8 and MCP-1 were increased obviously in LPS treated HPMECs. UFH might suppress LPS-activated NF-κB signaling pathway, contributing to the inhibitory effects of chemokines in HPMECs.

Objective To develop an HPLC method to determine the contents of chrysophanol, emodin, physcion, aloe-emodin, sennoside A, and sennoside B simultaneously in the aerial part of Rheum tanguticum Maxim.ex Balf. Methods The analysis was achieved with an Agilent ZORBAX SB-C18 analytic column (250 mm × 4.6 mm, 5 μm) by gradient elution of methanol-0.1％ phosphoric acid in water gradient elution system (0 min, 70％ B; 5 min, 65％ B;8–16 min, 60％ B; 18–23 min, 55％ B; 25–30 min, 45％ B; 32–39 min, 35％ B; 49–57 min, 24％ B; 65–72 min, 20％ B). The flow rate was 1.0 mL/min; detection wavelength was 254 nm; the column temperature was room temperature; the quantification used external standard method. Results The peak areas and concentrations of chrysophanol, emodin, physcion, aloe-emodin, sennoside A, and sennoside B showed good linear relationship within a certain concentration range (r≥0.9995); the RSD of precision was 0.75％–1.17％; the RSD of repeatability was 0.99％–2.06％; the RSD of stability was 0.97％–1.76％; the average recoveries were 99.7％–100.4％. The results showed that there were differences in content between the root and aerial part of Rheum tanguticum Maxim.ex Balf. Conclusion HPLC method for simultaneous determination of contents of 6 contents of the aerial part of Rheum tanguticum Maxim.ex Balf can be used as the references for quality control.

Background The primary pathological basis of diabetic retinopathy (DR) is new blood formation due to anoxia and inflammation,which results in breakdown of blood-retinal barrier (BRB).Vascular endothelial growth factor (VEGF) is a key factor promoting neovascularization.Researches determined that lycium barbarum polysaccharides (LBP) can protect cells against oxidative damage.However,the study of LBP in ophthalmology is lack.Objective This study was to investigate the effects of LBP on the dynamic pathological change of retinal vessels and expression of VEGF in retina of diabetic rats.Methods One hundred and seventeen SPF male SD rats were randomly divided into the normal control group,the diabetic mellitus (DM) group and the LBP group according to random number table.Type 1 DM models were induced by intraperitoneal injection of streptozotocin (STZ,55 mg/kg) in the rats of the DM group and the LBP group,and then 250 mg/kg LBP was intragastically administered in the rats of the LBP group.The morphological change of retinal vessels was dynamically observed by retinal stretched preparation with Evans blue (EB) in 4,10 and 16 weeks after modeling.E B (45 mg/kg) was slowly injection via jugular vein,and 1％ polyoxymethylene was infused into the left ventricule.The eyeballs were extracted and retina were isolated.EB content in the retinas (mg/g) was calculated using retinal stretched preparation method at the time points mentioned above.Expressions of VEGF protein and mRNA in the retinas were detected by immunohistochemistry and real-time quantitative PCR at various time points,respectively.Results Retinal stretched preparation with EB exhibited that the abnormal degree in the shape,diameter of vessels and leakage of the retinal blood vessels were significantly slighter in the LBP group than those of the DM group in 4,10,16 weeks after modeling.At 4,10,16 weeks,EB content in the retinas was (12.17±1.55),(16.46±1.60) and (19.55±1.49) mg/g,which was significantly lower than (15.76± 1.90),(21.61 ±2.05) and (26.30±2.28) mg/g of the DM group (P＜0.05).Immunochemistry showed that the expression of VEGF protein primarily located at retinal ganglion cells (RGCs) layer.The staining intensity for VEGF protein was weaker in the LBP group than that of the DM group.The expression levels of VEGF protein (A value) in the LBP group were 0.234±0.011,0.331±0.023 and 0.536±0.031at various time points,with significant decline in comparison with 0.281±0.018,0.533±0.055 and 0.765±0.075 of the DM group (all at P＜0.05).Real-time quantitative PCR revealed that the expression levels of VEGF mRNA were 0.157±0.013,0.505 ±0.114 and 1.577±0.074 in the LBP group at various time points,which were significantly lower than 0.235±0.209,1.043±0.084 and 2.446±0.061 of the DM group (all at P＜0.05).Conclusions LBP can alleviate the DM-induced retinal vasculopathy,lessen the leakage of vessels well,and further protect the BRB.

Objective:To establish an improved method for the determination of diethylene glycol and ethylene glycol in glycerol on the basis of the method in Chinese pharmacopoeia (2010 edition). Methods: Glycerol samples were extracted by methanol, and detected by gas chromatography with a flame ionization detector. The column was a DM-624 capillary column(30 m × 0. 53 mm,3μm), and the initial temperature was 80℃ in the first 6min and then risen to 100℃ at the speed of 50℃·min-1, kept for 8min,fi-nally risen to 220℃ at the speed of 10℃·min-1 , kept for 10min. The split injection was used with the split ratio of 100∶1. The in-jection temperature was 250℃ and the detector temperature was 280℃. The flow rate was 6. 0ml·min-1 . Results:The RSD(%) of the method was lower than 5%, and the resolution of all chromatographic peaks met the requirements of the pharmacopoeia. The meth-od was used to analyze the commercial product of glycerol, and the contents of diethylene glycol and ethylene glycol were both less than the specified limits. Conclusion:The method is simple and rapid, and suitable for glycerol impurity inspection.

Objective To analyze the correlation between liver tissue hypoxia inducible factor‐1α(HIF‐1α) protein expression and multidrug resistance protein (MRP) expression between .Methods Our hospital from March 2012 to March 2013 the Depart‐ment of Pathology of the liver paraffin‐embedded specimens of a total of 83 cases of specimen processing ,production of tissue sec‐tions ,HIF‐1αexpression was observed ,the positive expression of MRP .Results 83 cases of specimens in HIF‐1α 50 cases were positive ,the positive rate was 60 .2% ;of which the well‐differentiated tumor samples of HIF‐1α positive rate was 74 .1% ,signifi‐cantly higher than the 28 .0% poorly differentiated (P<0 .05);and the absence of lymph node metastasis positive rate of HIF‐1αwas no significant difference (P>0 .05) .In 83 cases of samples ,58 samples were MRP positive ,the positive rate was 70 .0% ;MRP positive rate among the high degree of 77 .6% ,significantly higher than the 52 .0% poorly differentiated (P<0 .05);and lymph node tumors MRP positive rate of metastasis were 76 .30 ,64 .4% respectively ,the difference was not statistically significant (P>0 .05) .MRP expression in the same samples were positive ,the HIF‐1α expression was also significantly increased .Conclusion HCC HIF‐1αprotein expression and MRP expression has some relevance .

Objective: To investigate the effect of thalidomide and triamcinolone acetonide (TA) on human umbilical vein endothelial cells (HUVEC). Methods: The primary culture of HUVEC with collagenase Ⅰ was identified with FⅧAg. There were 3 groups in the study including thalidomide group(10, 25, 50 and 75 mg/L), TA and thalidomide group and control group. The absorbance ratio was measured through MTT-test. Results: Both thalidomide group and TA and thalidomide group showed inhibitor effect on the growth of HUVEC significantly(P < 0.01). The inhibitory effect was dose dependent. Compared to thalidomide group, the inhibitory effect was more significant in TA and thalidomide group(P < 0.05). Conclusion: Thalidomide can inhibit the growth of HUVEC; however, the effect was more significant in TA and thalidomide group.