OBJECTIVE:To establish a capillary zone electrophoresis with high frequency conductance method for the determination of ursolic acid in Radix et Rhizoma Clematidis.METHODS:The separation of sample was performed on an uncoated fused silica capillary(60 cm?75 ?m ID,effective length:56 cm) with 1.2 mmol?L-1 triethylamine-HCl(pH 10.60) and 0.24 mmol?L-1 ?-cyclodextrin as running buffer solution.The separation voltage was 12.5 kV and the gravity injection of sample was conducted for 10 s at a height of 20 cm.RESULTS:The calibration curve of ursolic acid was linear over the concentration range of 5.55～111.0 mg?mL-1(r=0.999 3) and the average recovery for ursolic acid was 96.0%(RSD=1.39%,n=6). CONCLUSION:The method was proved to be simple,accurate,rapid and reproducible,and suitable for the quantitative determination of ursolic acid in Radix et Rhizoma Clematidis.

OBJECTIVE:To establish a RP-HPLC method for the determination of imperatorin in Qingwen jiedu tablets(heat-clearing and detoxifying agent).METHODS:The chromatographic separation was performed on Diamonsil-C18(250 mm?4.6 mm,5 ?m)with column temperature kept at 35 ℃.The mobile phase consisted of methanol-water(70∶30)at a flow rate of 0.8 mL?min-1 with detection wavelength set at 300 nm.RESULTS:The linear range of imperatorin was 2.7～27.0 ?g?mL-1(r=0.999 9)with its average recovery at 98.9%(RSD=1.44%,n=6).CONCLUSION:The established method was proved to be well-separated,sensitive,accurate and applicable for the quality control of Qingwen jiedu tablets.

OBJECTIVETo construct a vector containing protein transduction domain (PTD) and bcr/abl fusion gene of chronic myelogenous leukemia and express PTD-bcr/abl fusion protein in E. Coli.

METHODSDNA fragment encoding PTD was synthesized and fused to PCR-amplified bcr/abl gene fragment, then inserted into plasmid pET-16b to get the expression vector pEPb containing PTD-bcr/abl fusion gene, which was transfected and expressed in E. Coli LB21. PTD-bcr/abl fusion protein was purified by affinity chromatography.

RESULTS523 bp bcr/abl fusion gene was effectively amplified. The PTD-bcr/abl gene sequencing showed the same sequence as scheduled. The fusion peptide was successfully expressed in E. Coli and purified.

CONCLUSIONThe results may provide a new PTD-bcr/abl fusion peptide for the immunotherapy of CML.

Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; genetics ; Fusion Proteins, bcr-abl ; genetics ; metabolism ; Gene Expression ; Gene Products, tat ; genetics ; metabolism ; Genetic Vectors ; genetics ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; Recombinant Fusion Proteins ; genetics ; metabolism

Objective:To observe the changes of macular morphology and blood flow after minimally invasive vitrectomy (PPV) in patients with severe non-proliferative diabetic retinopathy (sNPDR).Methods:A prospective clinical study. From January 2020 to April 2021, 17 consecutive sNPDR patients with 17 eyes who were diagnosed and received PPV treatment at the Zhongshan Ophthalmic Center of Sun Yat-sen University were included in the study. There were 12 males with 12 eyes and 5 females with 5 eyes; the average age was 55 years old; the average duration of diabetes was 11 years; the average glycosylated hemoglobin was 7.9%. Before the operation and 1, 3, and 6 months after the operation, all the affected eyes underwent best corrected visual acuity (BCVA), standard 7-field fundus color photography, and optical coherence tomography angiography (OCTA). An OCTA instrument was used to scan the macular area of the affected eye with in the range of 3 mm×3 mm to measure the central subfoveal thickness (CST), the thickness of the ganglion cell complex (GCC) in the macular area, the thickness of the retinal nerve fiber layer (RNFL), and the superficial capillary plexus (SCP) vessel density and perfusion density in the macular area, macular avascular zone (FAZ) area, a-circularity index (AI). Before the operation and 6 months after the operation, the least significant difference test was used for the pairwise comparison.Results:Before the operation, 1, 3, and 6 months after the operation, the FAZ area of the macular area were 0.34±0.14, 0.35±0.10, 0.37±0.10, 0.36±0.13 mm 2, respectively; AI were 0.52±0.13, 0.54±0.11, 0.57±0.10, 0.60±0.11; CST was 282.6±66.7, 290.4±70.9, 287.2±67.5, 273.2± 49.6 μm; GCC thickness were 77.1±15.5, 74.3±13.9, 72.6±16.2, 78.5±18.3 μm; the thickness of RNFL was 97.9±13.8, 101.3±14.6, 97.7±12.0, 96.1±11.4 μm, respectively. The overall blood flow density of SCP in the macula were (16.79±1.43)%, (16.71±1.82)%, (17.30±2.25)%, (17.35±1.22)%; the overall perfusion density were 0.32±0.02, 0.32±0.03, 0.33±0.03, 0.33±0.02, respectively. After the operation, the CST increased first and then decreased; the thickness of RNFL increased 1 month after the operation, and then gradually decreased. Comparison of the parameters before and 6 months after the operation showed that the AI improved, and the difference was statistically significant ( P=0.049); the difference in FAZ area and the thickness of CST, GCC, and RNFL was not statistically significant ( P=0.600, 0.694, 0.802, 0.712); There was no statistically significant difference in the retina SCP blood flow density and perfusion density in the macular area ( P=0.347, 0.361). Conclusion:Compared with before surgery, there is no significant change in macular structure and blood flow density in sNPDR patients within 6 months after minimally invasive PPV.