Objective To investigate the effect of endovascular brachytherapy with Rhenium-188-filled catheter balloon on cell proliferation,phenotype of smooth muscle cell and secretion of transforming growth factor; to explore the possible mechanism of this method employed in restenosis prevention. Methods After the right common carotid arteries were overstretched by 3.5?20 mm balloon,twenty New Zealand White rabbits were randomized into irradiation (n=10) and control group (n=10). Endovascular bachytherapy was performed wth a 188[KG*2/2]Re filled 3.0?20 mm balloon catheter delivering a dosage of 16 Gy (0.5 mm underneath the luminal surface) in the irradiation group, whereas isovolume normal-sodium-filled balloon catheter was used in the control group for an equal period. After thirty days, analysis of the carotial arteries was made by measns of histological and immunohistochemical staining. Results (1) The positive staining index of proliferation cellular nuclear antigen (PCNA) and transforming growth factor (TGF) of the irradiation group is significantly less than the control group (0.028?0.005 vs 0.054?0.011, 0.033?0.008 vs 0.063?0.014, respectively); (2) The positive staining of ?-actin index in intima has no significant difference between the irradiatin group and the control group. Conclusion (1) The preventive effect on restenosis with endovascular brachytherapy is possibly achieved by suppression of cellular proliferation and transforming growth factor secretion. (2)The phenotype of smooth muscle cell is possibly unaffected by endovascular brachytherapy.

Objective To test the protective immunity in mice induced by recombinant Schistosoma japonicum Sj14FABP through several adjuvant formulations. Methods The recombinant Schistosoma japonicum Sj14FABP was prepared by expression in E. coli as a GST fusion protein (rSj14/GST) and used to vaccinate outbred Kunming mice by using complete Freund's adjuvant (FCA)/incomplete Freund's adjuvant (FIA), Bacillus Calmette-Guerin (BCG) and the immunostimulating complex (ISCOM) as adjuvant respectively. Results The purified recombinant protein rSj14/GST was immunogenic in mice, and 34.3% and 36.0% worm reduction rates were obtained in outbred Kunming mice immunized intradermally with BCG adjuvant and immunized subcutaneously with ISCOM adjuvant respectively, compared with non-vaccinated control group. However, intramuscularly vaccination with rSj14/GST in FCA/FIA was not protective, although the high level of IgG antibody was induced. Conclusion Both BCG and ISCOM are suitable adjuvants for rSj14/GST.

The gene fragment encoding the egg-shell precursor protein of Schistosoma japonicum was amplified with RT-PCR by using PCR primer designed according to the 423 bp cDNA fragment of the Philippine strain of S.japonicum, the corresponding mDNA fragment of Chinese strain as template and then the 5' and 3' ends of this gene cDNA were amplified with 5' RACE and 3' RACE by using a series of primers designed according to the result of sequencing. Result of sequence analysis showed that this fragment, named as Sj423, contained a complete open reading frame (ORF) of gene encoding the egg-shell precursor protein of S.japonicum.(Chinese strain). As demonstrated by sequencing analysis. No intron could be detected in this gene fragment. This gene was subsequently expressed in E.coli after cloning into the expression vector pET28c(+). The molecular mass of the expressed product of this gene was 20.9 kDa as revealed by SDS-PAGE analysis, and Western blot analysis showed that the recombinant protein expressed could react well with the rabbit antiserum against the worm antigen of S.japonicum;indicating the good antigenicity of this expressed product.

Objective To investigate the oxidative stress status and cardiac myocyte apoptosis in rats with propylthiouracil-induced hypothyroidism and to examine the effect of levothyroxine ( LT4 ),vitamin E ( VitE ),and both supplementation on this experimental model.Methods Male Sprague Dawley rats were divided into normal control group( NC),propylthiouracil group( PTU ),LT4 treatment group( PTU + LT4 ),VitE treatment group ( PTU +VitE),LT4 and VitE combination therapy group (PTU + LT4 + VitE).Serum T3,T4,TSH levels,serum and myocardial superoxide dismutase ( SOD ) activity and malondialdehyde ( MDA ) content were determined.Cardiac myocyte apoptotic index was made with TUNEL.Results ( 1 ) Compared with NC group,T3 and T4 levels were significantly lower and TSH level was significantly higher in PTU group and PTU+VitE groups( P＜0.05 ).Compared with PTU group,T3 and T4 levels were significantly higher and TSH level was significantly lower in PTU + LT4 group and PTU + LT4 + VitE group( P＜0.05 ).There were no statistical differences in T3,T4,and TSH levels between PTU group and PTU + VitE group( P＞0.05 ).( 2 ) Compared to NC group,serum and myocardial MDA levels in PTU group increased significantly( P＜0.05 ),serum SOD activity decreased significantly ( P＜0.05 ),while myocardial SOD activity showed no significant difference( P＞0.05 ).( 3 ) Compared with NC group,myocardial apoptosis index was significantly higher in PTU and PTU + VitE groups( P＜0.05 ).Compared with PTU group,myocardial apoptosis index was significantly lower in PTU + LT4 group and PTU + LT4 + VitE group( P＜0.05 ).Conclusion LT4,VitE,and their combined treatment of hypothyroidism in rats reduce the myocardial cells apoptosis,which may be related to the improvement of thyroid function and amelioration of oxidative stress.

Objective · To design and synthesize five new tropane compounds, and test their antagonistic activity against M3 receptor and inhibition activity to neutrophil elastase (NE), of which the structure-activity relationship were preliminarily investigated. Methods · The five compounds, A1-A3,B1 and C1, were prepared with 3α-hydroxy-tropane (A0) as the starting material by modifying the structure in C-3α position and N atom on the tropane skeleton. The antagonistic activity of the compounds to muscarinic M3 receptors on tracheal rings of guinea pigs was evaluated by functional assays in vitro. The hydrolysis of PGlu-Pro-Val-PNA as substrate was catalyzed by NE to get colorful nitroaniline (PNA). The NE inhibition activity of the tropane compounds was obtained by determining the absorbance [(D(405 nm)] of PNA. Results · The five new tropane compounds generated strong antagonistic activity against M3 receptors. Among them, A2 had the greatest activity [antagonistic parameter pA2(M3)=9.004], and elicited obvious inhibitory effect to NE (inhibition ratio YA2=20.29%). Conclusion · Introducing strong electron-attraction group, such as sulfuryl and hydrophobic group with large volume into C-3α position on the tropane skeleton can improve the M3 receptor antagonistic activity as well as the NE inhibition activity.

Objective To screen protective antigen genes and construct the T7 phage display library from adult worms of Schistosoma japonicum.Methods Total RNA was extracted from adult worms of S.japonicum by Trizol reagent and mRNA was isolated from the total RNA.The ds cDNA was synthesized by reverse transcription using random primer.Directional EcoRⅠ/HindⅢ linkers were ligated into the ends of ds cDNA and the ds cDNA was digested with EcoRⅠand HindⅢ,which resulted in ds cDNA with EcoRⅠand HindⅢ adhering ends.The digested ds cDNA fragments longer than 300 bp in length were fractionated and ligated into T7 Select 10-3b vector.After packaging in vitro,the T7 Select 10-3b vector was transformed into BLT5403 to construct the T7 phage display cDNA library.Plaque assay and PCR were used to evaluate the library.Seven known objective genes of S.japonicum were screened by PCR to detect the representation of the library.Result Primary library capacity was 4.98?106 pfu,and the titer of amplified library was 3.85?1011 pfu/mL.The PCR identification result of 96 clones picked at random showed that recombination rate was 93.8%,in which 95.6% inserted cDNA fragments were longer than 300 bp in length.All the seven known objective genes of S.japonicum were amplified from the library.Conclusion The T7 phage display library from adult worms of Schistosoma japonicum was constructed.

Objective To construct a T7 phage display cDNA library from the lung of Microtus fortis for further screening the schistosomiasis-resistence-related genes of Microtus fortis. Methods mRNA was isolated from total RNA extracted from the lungs of Microtus fortis by TRIzol reagent, and was used to synthesize double strain cDNA by the reverse transcription. Then the double strain cDNA was given with EcoRⅠ and Hind Ⅲ adhering ends by ligation with the directional EcoRⅠ/Hind Ⅲ linkers and digestion with EcoRⅠ and Hind Ⅲ. The double strain cDNA fragments longer than 300 bp in length were fractionated by the Mini Column, and ligated into the T7 Select 10-3b vector with EcoRⅠ and Hind Ⅲ adhering ends. After packaging in vitro, the recombinant T7 Select 10-3b was transformed into BLT5403 to construct a T7 phage display cDNA library. Results The library constructed here contained 1.5?106 clones and the titer of the amplied library was 1.1?1012 pfu/ml. The PCR identification results of 100 clones picked at random showed that 91% clones were recombinant and 90% of recombinant clones contained cDNA fragments longer than 300 bp in length. Conclusion A T7 phage display cDNA library from the lung of Microtus fortis is successfully constructed.

To examine the feasibility of using a computer tool for stratifying the severity of Coronavirus Disease 2019 (COVID-19) based on computed tomography (CT) images. We retrospectively examined 44 confirmed COVID-19 cases. All cases were evaluated separately by radiologists (visually) and through an in-house computer software. The degree of lesions was visually scored by the radiologist, as follows, for each of the 5 lung lobes:0, no lesion present;1,<1/3 involvement;2,>1/3 and<2/3 involvement;and 3,>2/3 involvement. Lesion density was assessed based on the proportion of ground-glass opacity (GGO), consolidation and fibrosis of the lesions. The parameters obtained using the computer tool included lung volume (mL), lesion volume (mL), lesion percentage (％), and mean lesion density (HU) of the whole lung, right lung, left lung, and each lobe. The scores obtained by the radiologists and quantitative results generated by the computer software were tested for correlation. A Chi-square test was used to test the consistency of radiologist- and computer-derived lesion percentage in the right/left lung, upper/lower lobe, and each of the 5 lobes. The results showed a strong to moderate correlation between lesion percentage scores obtained by radiologists and the computer software (r ranged from 0.7679 to 0.8373, P < 0.05), and a moderate correlation between the proportion of GGO and mean lesion density (r=-0.5894, P<0.05), and proportion of consolidation and mean lesion density (r=0.6282, P<0.05). Computer-aided quantification showed a statistical significant higher lesion percentage for lower lobes than that assessed by the radiologists (x2 = 8.160, P = 0.004). Our experiments demonstrated that the computer tool could reliably and accurately assess the severity and distribution of pneumonia on CT scans.