Objective: To establish a method for determination of tetrahydropalmatine in Anwei tablets. Methods: tetrahydropalmatine in Anwei Tablets was extracted by chloroform and determined by dual wavelength TLC scanning (? s=275nm,? R=340nm).Results: The average recovery was 95%.17%, RSD was 2.50% (n=5). The contents of tetrahydropalmatine in three batches were 7.45,7.17 and 7.90?g/tablet, respectively.Conclusions: The method is stable and feasible.

Objective To evaluate the diagnostic value of multi-slice spiral CT(MSCT)perfusion imaging in chronic obstructive pulmonary disease(COPD).Methods Twenty COPD patients and20 volunteers underwent 8-row detector spiral CT(MSCT)perfusion imaging using cine scan mode with5 mm slice thickness.0.5 s rotation time and a total scan time of 45 s with 5 s intervals.60 ml contrast agent(300 mg I/ml)were administered at a rate of 4 ml/s from the forearm superficial vein.The imaging data were transferred to a workstation.A time-density curve and pseudo-color map were generated automatically with GE CT perfusion 3 software,the blood flow(BF),blood volume(BV),mean transit time(MTr)and permeability surface(PS)were measured.Results Time-density curve was flatter and the peak of the curve was obviously lower in COPD patients than the volunteers.The BF.BV.PS in COPD volunteers was(10.58 ±4.85)s and(4.50 ±1.71)s respectively.The BF,BV and PS in COPD patients Was lower than the volunteers,the MTY was higher(P＜0.01).Conclusion MSCT perfusion imaging is helpful for the diagnosis of COPD.

Objective To study the image quality and radiation dose in dual-source CT cerebral angiography with prospective ECG-triggered sequence mode (step-and-shoot,SAS).Methods A total of forty-three patients with clinically suspected cerebral vascular disease underwent cerebral CT angiography with prospective ECG-triggering (step-and-shoot,SAS).Data acquisition was at 60％ R-R interval of the ECG presentation mode.The post-processing included maximum intensity projection (MIP),multiplanar reformation (MPR) and volume rendering (VR).The CTA image quality,radiation dose and rates of excellent images were evaluated.Results The CTA image quality score was 4.72 ± 0.50 and 97.7％ (42/43) patients had excellent CTA images.The average effective dose of SAS-CTA was (0.22 ± 0.01 )mSv,which was lower by 76.31％ than that of DE-CTA.Conclusions Prospective ECG-triggering sequence could be used in cerebral angiography with a significant reduction in radiation dose and diagnostic image quality.

Objective To compare the quality and radiation doses of coronary artery angiography under the natural heart rate condition between Flash spiral heart mode and prospective electrocardiogramtriggering sequence mode using dual-source,in order to choose personalized low doses of coronary artery scanning mode.Methods Sixty patients who underwent coronary angiography(CTA)on a 128-slice,dualsource CT scanner were divided into 2 group i.e,group A(27cases)and group B(33 cases).Flash spiral heart scan mode was employed for group A.Inclusion criteria included:heart rate<65 bpm.regular sinus rhythm,heart rate fluctuation less than ±5 bpm.Date acquisition was set at 60% of the R-R interval.Prospective electrocardiogram-triggering sequence scan mode(SAS)was performod for group B.Inclusion criteria included:(1)heart rate≥65 bpm,(2)arrhythmias,premature beat,fibrillation atrial.Exclusion criteria included:bad holding breath.Date acquisition(1)At low heart rate(≤75 bpm),date acquisition was set at 60%-80%of the R-R interval.(2)At high heart rate(>75 bpm),date acquisition was set at 30%-50%of the R-R interval. (3)At the arrhythmias,premature beat,fibrillation atrial,date acquisition was set at 20%-90%of the R-R interval.In both gronps,patients with a BMI≥25.0kg/m2 were examined with a tube voltage of 120 kV.while the other patients with a BMI<25.0 kg/m2 were examined with a tube voltage of 100 kV.The BMl was(24.6±1.0)kg/m2 in group A,while that was (24.6±0.9)kg/m2 in group B.In both groups,all images were transferred to the workstation for further processing and analysis.The imaging quality of coronary artery segments and the radiation dose were compared with t test.Results A total of 336 coronary artery segments were evaluated in group A and 412 segments were evaluated in group B.The imaging quality of coronary artery segments were scored.Excellent or good was achieved in 98.2%(330 of 336)artery segments in group A,and that was 98.1%(404 of 412)in group B.There was no statistical difference in imaging quality between the two groups(t=0.513,P=0.608).The average effective dose was(0.74±0.29)mSv in group A,whereas that was(3.67±1.37)mSv in group B.There was a significant difference between the two groups(t=-10.858,P=0.000).Conclusions The personalized low doses coronary artery scanning mode can substantially reduce radiation damage while preserving good imaging quality.

Objective:To determine the expression of BMP4 in hepatocellular carcinoma (HCC) and to study the role of BMP4 in inducing epithelial-mesenchymal transition (EMT) to analyze the effect of BMP4 on the migration and invasion of HCC cells. Methods: The expression of BMP4 in HCC specimens was examined by immunohistochemistry staining, and the correlations were analyzed between the expression of BMP4 and clinicopathological data. The BMP4 expression plasmid was transfected into HepG2 cells to induce exogenous overexpression of BMP4 protein. The changes of HepG2 cell morphology were detected after BMP4 transfection by using a microscope; the changes of the expression of BMP4, EMT-related protein (E-cadherin, Vimentin) in HepG2 cells were detected by Western blot after transfection of BMP4;the wound healing assay in vitro was used to detect the effects of BMP4 gene transfection on the ability of migration of HepG2 cells;the invasion assay was used to determine the role of transfection of BMP4 on the invasive potential of HepG2 cells. Results: Immunohistochemistry staining method displayed that BMP4 expression was positively associated with age, histological differentiation, stage, and poor prognosis. After BMP4 overexpression, the morphology of HepG2 cells showed significant changes from a paving stone structure with cell-cell adhesion to a fibroblastic shape, which showed typical EMT change; Western blot exhibited that the expression of E-cadherin was downregulated and the Vimentin expression was upregulated in HepG2 cells;the wound healing and invasion assay showed that the migration and invasion potentials of HepG2 cells were significantly enhanced. Conclusion: BMP4, which displayed a high expression in HCC specimens, was closely associated with clinicopathologic data, and BMP4 may promote migration and invasion of HCC cells by inducing epithelial-mesenchymal transition.

Objective To investigate the effect of fluoride exposure on expression of miRNA (miR)-200c and its target in human osteoblast Saos-2 cells.Methods Saos-2 cells were cultured in DMEM/F-12 medium and treated with fluoride (sodium fluoride,NaF).There were two groups including:control group (0 mg/L) and fluoride group (4 mg/L).Cells were harvested after 48 hours of culture with fluoride.The expression of miR-200c,the mRNA of alkaline phosphatase (ALP),osteocalcin (BGP),the target phosphatase and tensin homolog deleted on chromosome ten (PTEN) and dual-specific phosphatase 1 (DUSP1) of miR-200c was detected by qRT-PCR.The protein expression of PTEN and DUSP1 was detected by Western blotting.Results The expressions of ALP,BGP mRNA and miR-200c in Saos-2 cells in the fluoride group (23.60 ± 1.87,9.41 ± 0.94,8.61 ± 0.26) were higher than those in the control group (1.00 ± 0.11,1.00 ± 0.07,1.00 ± 0.12).The differences were statistically significant (t =-24.084,-18.388,-8.687,P ＜ 0.05).The mRNA expressions of PTEN and DUSP1 in the fluoride group (0.63 ± 0.02,0.38 ± 0.02) were lower than those in the control group (1.02 ± 0.24,1.02 ± 0.24).The differences were statistically significant (t =3.327,5.454,P ＜ 0.05).The protein expressions of PTEN and DUSP1 in Saos-2 cells in the fluoride group (1.19 ± 0.10,0.83 ± 0.07) were lower than those in the control group (1.81 ± 0.14,1.44 ± 0.25).The differences were statistically significant (t =6.250,4.171,P ＜ 0.05).Conclusion Exposure to fluorine may increase the expression of miR-200c in Saos-2 cells,and fluorine may act on PTEN and DUSP1 through miR-200c,downregulates the mRNA and protein expression levels of PTEN and DUSP1.

Objective:To investigate the regulatory effect of transient receptor potential cation channel subfamily C member 3 (TRPC3) on the retina in oxygen-induced retinopathy (OIR) mice and biological behavior of human retinal vascular endothelial cells (HREC).Methods:A total of 32 healthy SPF grade 7-day-old C57BL/6 mice were selected and randomly divided into a control group and an OIR group by the random number table method, with 16 mice in each group.The control group received no special treatment, and the OIR model was established in the OIR group.On postnatal day 17 (PN17), the success of the model establishment was verified by immunofluorescence staining of the retinal patch.The in vitro cultured HREC were divided into a normal control group, a transfection reagent group, and a si-TRPC3 group.The normal control group received no special treatment, while the transfection reagent group and the si-TRPC3 group were transfected with transfection reagent or transfection reagent + si-TRPC3.The relative expression of TRPC3 mRNA was detected by real-time quantitative fluorescence PCR.The relative expressions of TRPC3, transcription factor NF-E2 related factor (Nrf2), and superoxide dismutase (SOD) proteins were determined by Western blot.HREC were further divided into a normal control group, a vascular endothelial growth factor (VEGF) group, a si-TRPC3 group, and a Pyr3 (TRPC3 channel inhibitor) group, which were cultured in complete medium, medium containing 20 ng/ml VEGF recombinant protein, medium containing 20 ng/ml VEGF recombinant protein (si-TRPC3 transfection for 72 hours), and medium containing 20 ng/ml VEGF recombinant protein+ 1 μmol/L Pyr3 for 48 hours, respectively.The proliferation ability of HREC was detected using cell counting kit 8 (CCK-8). The horizontal and vertical migration ability of cells were detected by cell scratch assay and transwell assay, respectively.This study followed the 3R principles of animal welfare and was approved by the Ethics Committee of Hebei Eye Hospital (No.2023LW04). Results:Pathological neovascular clusters with strong fluorescent staining appeared in the retina of OIR mice on PN17.The relative expressions of TRPC3 mRNA and protein in the retina of OIR mice were 2.057±0.244 and 1.517±0.290, respectively, significantly higher than 0.983±0.033 and 0.874±0.052 of control group ( t=6.165, 3.094; both at P<0.05). The relative expression levels of TRPC3 mRNA and protein were significantly lower, and the relative expression levels of Nrf2 and SOD proteins were higher in the si-TRPC3 group than in the normal control and transfection reagent groups, and the differences were statistically significant (all at P<0.05). The CCK-8 experiment results showed that the cell absorbance value was higher in the VEGF group than in the normal control group, and lower in the si-TRPC3 and Pyr3 groups than in the VEGF group, with statistically significant differences (all at P<0.05). The results of the cell scratch experiment showed that the lateral migration rate of VEGF group cells was higher than that of normal control group, while the lateral migration rate of si-TRPC3 group and Pyr3 group cells was lower than that of VEGF group, and the differences were statistically significant (all at P<0.05). The transwell experiment results showed that the number of stained cells in the VEGF group was higher than that in the normal control group, and the number of stained cells in the si-TRPC3 group and Pyr3 group was lower than that in the VEGF group, with statistically significant differences (all at P<0.05). Conclusions:Hypoxia induces increased TRPC3 expression in OIR mouse retina, and downregulation of TRPC3 inhibits HREC proliferation and migration.The mechanism is related to the activation of the Nrf2-related oxidative stress pathway.

Objective To study the effect of T-2 toxin on proliferation and cell cycle of rat chondrocytes,in order to provide a new idea in molecular mechanism of T-2 toxin-induced chondrocyte damage.Methods Primary chondrocytes of neonatal Wistar rats were isolated and stained by toluidine blue staining and type Ⅱ collagen immunofluorescence staining.The effects of different concentrations of T-2 toxin [0 (control),1,5,10,20,50,100 μg/L)] on proliferation of chondrocytes for 24 h were detected by cell counting kit-8 (CCK-8) method,and control,1 (low dose),5 (medium dose),and 10 μg/L (high dose) T-2 toxin were selected for subsequent experiment;cell cycle changes were detected by flow cytometry;Real-time PCR and Western blotting were used to detect the effects of T-2 toxin on mRNA and protein expressions of proliferating cell nuclear antigen (PCNA) and Cyclin D1 in chondrocytes.Results With increase of T-2 toxin concentration (control,1,5,10,20,50,100 μg/L),the cell survival rates [(100.00 ± 0.00)％,(93.12 ± 1.66)％,(77.12 ± 1.11)％,(59.44 ± 4.09)％,(46.64 ± 3.86)％,(38.15 ± 3.37)％,(33.79 ± 0.99)％] were decreased,and the differences were statistically significant (F =139.21,P ＜0.05).The percentages of quiescent phase/pre-DNA synthesis phase (G0/G1 phase) ceils in 1,5,10 μg/L T-2 toxin groups [(22.03 ± 0.42)％,(30.54 ± 2.61)％,(36.01 ± 1.51)％] were significantly higher than that in control group [(13.79 ± 1.65)％,P ＜ 0.05];the percentages of DNA synthesis phase (S phase) cells [(60.27 ± 3.53)％,(53.88 ±4.38)％,(49.55 ± 2.49)％] were significantly lower than that in control group [(76.72 ± 4.24)％,P ＜ 0.05].The differences of mRNA levels of PCNA and Cyclin D1 between groups were statistically significant (F =46.80,17.97,P ＜ 0.05),and 5,10 μg/L T-2 toxin groups (0.77 ± 0.13,0.79 ± 0.08,0.60 ± 0.07,0.56 ± 0.05) were lower than the control group (0.99 ± 0.02,1.01 ± 0.01,P ＜ 0.05).The expressions of PCNA protein in 5,10 μg/L T-2 toxin groups (0.69 ± 0.03,0.49 ± 0.03) were lower than that in control group (0.92 ± 0.05,P ＜ 0.05);the expressions of Cyclin D1 protein in 1,5,10 μg/L T-2 toxin groups (0.80 ± 0.06,0.60 ± 0.07,0.33 ± 0.13) were lower than that in control group (0.95 ± 0.07,P ＜ 0.05).Conclusion T-2 toxin can inhibit the proliferation of chondrocytes,which may be worked through influencing the expression of cell cycle protein,causing cell cycle arrest,thereby inhibiting DNA synthesis.