1.Construction of pEGFP-N1-TGF-β1 recombinant plasmid and transfected it into primary cultured AEC-Ⅱ of neonatal piglet using lipofectamine 2000
Chinese Journal of Emergency Medicine 2016;25(1):50-56
Objective To construct pEGFP-N1-TGF-β1 recombinant plasmid and transfect it into primary cultured neonatal piglet type Ⅱ alveolar epithelium cell (AEC-Ⅱ) by using lipofectamine 2000,in order to provide basis of methodology for producing recombinant plasmids for transplantation of transfected AEC-Ⅱ into ALI/ARDS animal model lungs.Methods PCR primers were designed to amplify the human TGF-β1 cDNA fragment from plasmid.XhoI and EcoRI were used for double digesting the empty plasmid pEGFP-N1 and cDNA fragment of human TGF-β1.Then the products of double enzyme digestion by using T4 DNA ligase were connected and transformed into DH5α and cultured over night for 16 hours.The structure of recombinant plasmid was identified by using PCR and base sequencing to verify the correctness of pEGFP-N1-TGF-β1 recombinant plasmid.It was then transfected into primarily cultured AEC-Ⅱ by lipofectamine2000 mediated transfection and cultured for another 48 hour.Plasmid DNA (pEGFP-N1-TGF-β1 recombinant plasmid) and lipofectamine 2000 were added into serum-free DMEM respectively,then DNA suspension and Lipofectamine 2000 suspension were blended together and added into cells.After 24-48 hours later,the expression level of enhanced green fluorescent protein (EGFP) was evaluated under fluorescence microscope.Results The structure of vector was verified as pEGFP-N1-TGF-β1 recombinant plasmid by using PCR and base sequencing.Green fluorescence found in some cells showed that the pEGFP-N1-TGF-β1 recombinant plasmids had been successfully transfected into primary cultured AEC-Ⅱ,however,the transfection efficiency still need tobe further improved such as repeating the transfection procedure once again or using adenovirus mediated transfection method to improve the efficiceny and to transplant the cells into animal lungs eventually.Conclusions pEGFP-N1-TGF-β1 recombinant plasmid was successfully constructed and,for the first time,transfected into primarily cultured AEC-Ⅱ of newborn piglets.This established method should be useful for investigation of therapeutic effect and outcomes of lung with experimental acute lung injury.
2.Significance and ways of cultural integration in hospital regrouping
Bo YAN ; Hengjin DONG ; Yuanzhao XU
Chinese Journal of Hospital Administration 1996;0(07):-
In the course of hospital regrouping it is imperative to integrate medical functions, resource elements and hospital cultures. Yet the integration of hospital cultures plays a pivotal role, for the culture of a hospital is the basis of its survival and development and cultural integration can lead to the fusion of the staffs feelings, values and concepts and to the coordinated development of the hospitals hardware and software, which will produce greater benefits. Ways of cultural integration include: ①doing a good job of intention surveys and ideological persuasion prior to hospital regrouping; ②adopting different forms of cultural fusion for different forms of regrouping: with unbalanced regrouping, the cultures of the weaker hospitals should not be totally negated and attention should be paid to equality and impartiality, whereas with balanced regrouping, efforts should be made to enhance communications rather than to differentiate the primary and the secondary; ③giving full play to the role of informal organizations within the hospital in the fusion of different cultures.
3.Effects of ?-lipoic acid on the micro-and ultramicrostructure of the kidney in type 2 diabetic Goto Kakisaki rats
Xinfeng YAN ; Bo FENG ; Lei XU
Chinese Journal of Practical Internal Medicine 2006;0(23):-
Objective To evaluate the effect of ?-lipoic acid(ALA)as a antioxidant on the histomorphologic changes of the kidney in type 2 diabetic Goto Kakisaki rats.Methods Four healthy Wistar rats(normal control group)and 10 GK rats were divided randomly into two groups:diabetic control group and lipoic acid-treatment diabetic group(?-lipoic acid 35 mg/kg intraperitoneal injection qd alt),who were both fed for 12 weeks.Indexes of oxidative stress of kidney were measured.Sections of renal tissue were evaluated using both a light and electron microscope to assess histomorphologic changes.Paraffin sections of the left kidneys were prepared and stained with H&E,PAS and Masson stains to compare the histomorphologic changes.The image analysis software of Motic Image Advanced 3.1 was used to measure the area of glomerulus and mesangial region,and the thickness of the glomerular basement membrane(GBM).Results Compared with normal control group,there were significant decreases in GSH,SOD,VitC and VitE levels in diabetic control group.The kidneys of the normal control group exhibited normal cortical morphology as determined by light microscope.Mild glomerulosclerosis,characterized by widening of the mesangial region,mesangial expansion and intercapillary cells proliferation,and GBM thickening under electron microscope were observed in the diabetic control group.In the ALA-treatment group,all the above pathological changes were alleviated.Conclusion In the kidney of type 2 diabetic rat,there exists oxidative stress which is associated with diabetic nephropathy.?-Lipoic acid can alleviate the abnormal histomorphologic changes of the kidney via antioxidative effects.
4.Overview of a positive study of the feasibility and model of the restructuring of two second-tier hospitals
Bo YAN ; Hengjin DONG ; Yuanzhao XU
Chinese Journal of Hospital Administration 1996;0(02):-
Objective To find out universal problems in restructuring and explore relevant methodologies so as to provide the government with decision-making basis. Methods The method of partial quantification from statistics and operational research and the qualitative method from sociology were employed. Results Hospitals A and B assumed the functions of a second-tier hospital and satisfied the basic medical needs of District Z. With Hospital A being superior to Hospital B and inpatient service superior to outpatient service, the obstruction to the restructuring lay in the staffs of the two hospitals. If the restructuring should turn out to be successful, there would be more space for the expansion of their share of the medical market. The focus of the restructuring should be put on the advancement of technologies and the improvement of quality and efficiency. In particular, the basic medical needs of the residents in the areas concerned must be met. Conclusion In restructuring two hospitals, the model of loose cooperation and steady combination is recommended in the first place while the government ought to play the role of providing guidance, coordination and policy support.
5.Clinical study of facial nerve dissection approaches in parotidectomy
Xu YAN ; Yan PENG ; Haidong WEI ; Bo PENG ; Dawei MI
Journal of Practical Stomatology 2016;32(1):129-131
40 cases of parotidectomy were reviewed.Anterograde approach(group A)and retrograde approach(group B)for facial nerve dissection were used in 1 7 and 23 cases respectively.1 patients in group A and 8 in group B presented symptoms of temporary facial nerve paralysis(P <0.05).The anterograde method is safer and quicker.
6.Synthesis, characterization and molecular dynamics simulation of layered double hydroxides intercalated with aspartic acid
Yan SHEN ; Guoxiang PAN ; Bo XU ; Minhong XU
Journal of China Pharmaceutical University 2025;56(3):329-335
Traditional experimental methods are insufficient in the study of layered double hydroxides (LDHs) supramolecular structure and hydration expansion performance, and information on interlayer anionic arrangement and structural water molecules cannot be obtained. Aspartic acid intercalated magnesium aluminum hydrotalcite was synthesized using coprecipitation and ion exchange. The structure of hydrotalcite precursor and its aspartic acid composite materials was characterized by X-ray powder diffraction, differential thermal analysis, and infrared spectroscopy, and Materials Studio software was used to simulate the molecular dynamics of microstructure and hydration properties of LDHs intercalated with the aspartic acid drug. The prepared composite material had a regular layered structure and a single crystal phase. After intercalation with aspartic acid, the interlayer spacing increased from 0.84 nm to 1.13−1.17 nm; after intercalation, the thermal decomposition temperature of aspartic acid increased from 249 °C to 334 °C, greatly improving its thermal stability. The interlayer spacing of the intercalated hydrotalcite obtained from the experiment was close to the molecular dynamics simulation results when Nw=3−4. As more water molecules were inserted between the layers, the greater the interlayer distance became. Hydration energy increased gradually and tended to a certain value. The total number of hydrogen bonds increased gradually, the hydrogen bonds between laminates and anions decreased gradually, but the hydrogen bonds between laminates and water molecules increased gradually. The simulation results are close to the experimental results, which can lay a foundation for the design and synthesis of LDHs-based drug composites.
7.Report of a case of sphenoid sinus meningoencephaloceles misdiagnosed as cysts.
Yan-hong XU ; Ding-qing HUANG ; Hai-bo YANG
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2010;45(5):430-430
Diagnostic Errors
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Encephalocele
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diagnosis
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Female
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Humans
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Meningocele
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diagnosis
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Middle Aged
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Mucocele
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diagnosis
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Sphenoid Sinus
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pathology
8.The function of miR-193b in hepatocellular carcinoma
Haijiang YAN ; Wei FENG ; Bo ZHAI ; Haopeng ZHANG ; Lishan XU
Cancer Research and Clinic 2016;28(1):11-14
Objective To investigate the significance of miR-193b to biological behaviors of hepatocellular carcinoma (HCC). Methods 48 cases of HCC specimens and corresponding adjacent tissues were collected, and the miR-193b expression levels in these specimens were measured by real-time quantitative PCR. The miR-193b expression was measured by the same way in HepG2 and SMMC-7721 cells. The HepG2 and SMMC-7721 were transfected with miR-193b mimics or negative control miRNA mimic with Lipofectamine 2000, and the non-transfected cells were taken as blank control. The proliferation ability of the HCC cells were detected by MTT method, and the apoptosis rate was tested by flow cytometry. Results The expression level of miR-193b in HCC tissues (2.441 ±0.569) was significantly lower than that in the corresponding adjacent tissues (15.488±4.326) (P < 0.05). Compared with normal liver cell line L-O2, the expression levels of miR-193b were significantly lower in HepG2 and SMMC-7721 cells. Transfected with miR-193b mimic, the proliferation ability of HepG2 and SMMC-7721 cells were reduced, while their apoptosis were increased. Conclusion miR-193b may be negative to regulate the proliferation of HCC and increase its apoptosis.
9.Vertebral artery hypoplasia and its clinical significance
Shuangshuang YANG ; Yan JI ; Bo SONG ; Yuming XU ; Shilei SUN
International Journal of Cerebrovascular Diseases 2015;(3):209-213
Vertebral artery hypoplasia is a congenital vessel variation. Its incidence is from 1. 9 to 26. 5% . In recent years, studies have shown that vertebral artery hypoplasia may be a potential risk factor for posterior circulation infarction, especialy when it coexists with other cerebrovascular risk factors. Vertebral artery hypoplasia may also cause regional hypoperfusion and complex neurovascular regulation, and it also has a certaln link with migralne.
10.Quantitative determination of paeoniflorin and puerarin in Rg soft-exact by HPLC
Yan LIU ; Shunhang LIU ; Jiali JIA ; Bo XU
Chinese Traditional Patent Medicine 1992;0(06):-
AIM:This paper described a HPLC method.Paeoniflorin and puerarin in Rg soft-exact(Radix Puerariae lobatae,Redix Paeoniae alba,etc.) were simultaneously determined.METHODS:A simplified HPLC procedure was developed.A 4.6 mm?250 mm Alltima C_(18) column with a mobile phase consisting of NaH_2PO_4(20 Mm,pH 2.4)-Acetonitrile:Tetrahydrofuran(900:120:5) was used.The flow rate was 1.00 mL/min.It was detected at 320 nm.RESULTS:Paeoniflorin and puerarin were well separated.The average recoveries of paeoniflorin and puerarin were 98.10% and 101.2% respectively.CONCLUSION:The result shows that this method is convenient,rapid and accurate.