ObjectiveTo construct and identify a recombinant adenovirus expressing S protein receptor binding domain(RBD)and N protein of severe acute respiratory symptom coronavirus 2(SARS-CoV-2)Delta variant.MethodsThe RBD and N gene fragments of SARS-CoV-2 were cloned into pcDNA3.0BA vector respectively to construct recombinant plasmid pcDNA3.0BA-RBD-N. The RBD-CMV-N fragment was amplified by PCR and inserted into shuttle vector pShuttle-CMV. The shuttle plasmid pShuttle-RBD-N was then homologously recombined with pAdeasy-1 to obtain recombinant plasmid pAdeasy-1-RBD-N,which was transfected into HEK293 cells for recombinant adenovirus Ad-RBD-N packaging. The transcription of RBD and N genes of recombinant adenovirus in HEK293 cells was detected by RT-PCR,while the expre-ssion of RBD and N proteins by Western blot and immunofluorescence assay. 12 female BALB/c mice were immunized with Ad-RBD-N by intramuscular injection at a dose of 5 × 109copies per mouse. Blood samples were collected 14 d after immunization,and the serum antibody titers were measured by ELISA.ResultsThe RBD and N genes of recombinant adenovirus were transcribed normally in HEK293 cells,and the RBD and N proteins were expressed normally in MA104 cells. Mice immunized with the recombinant adenovirus produced specific IgG antibodies against RBD and N proteins.ConclusionThe recombinant adenovirus expressing S protein RBD and N protein of SARS-CoV-2 Delta variant was succe-ssfully constructed,which laid a foundation of the follow-up research on Delta variant vaccines.

Abstract?AIM:To investigate the therapeutic effect of Lucentis in the treatment of anterior segment neovascularization ( ASNV ) induced by central retinal vein occlusion ( CRVO) .?METHODS: This was a retrospective case series study for patients with ASNV secondary to CRVO from January 2013 to December 2014 and treated with intravitreal injection of lucentis. Best visual acuity ( BCVA ) , intraocular pressure ( IOP ) , iris examination and gonioscopy, and if necessary, fluorescein angiography and optical coherence tomography, were recorded.The follow-up time was 6-13mo, the average was 9.1 ± 2.9mo.?RESULTS: Eighteen patients ( 18 eyes ) were treated with intravitreal injection of lucentis;15 patients (15 eyes) of the 18 were treated with panretinal photocoagulation;the other 3 patients 3 eyes were not received the photocoagulation because of vitreous opacity. One patient was treated with glaucoma valve implantation.Six patients with only neovascularization and without glaucoma were improved in visual acuity whose intraocular pressure was controlled after combined treatment.Of the patients with neovascular glaucoma ( n=12 ) , after intravitreal injection of lucentis and panretinal photocoagulation, the IOP was controlled in 4 patients (4 eyes);the IOP of another 7 cases reduced but glaucoma drugs were still needed.One patient ( 1 eye ) received glaucoma valve implantation because of bad control on IOP with worse vision after implantation.The rubeosis disappeared in all patients.?CONCLUSION:Intravitreal injection of lucentis can stop neovascularization and help to control IOP.In the early stage without neovascular glaucoma, lucentis has better effects, which means early detection and intervention are important.

0.05). However, the MDA level in the experimental group was higher than that of the control on day 3 [(55.92?5.53)nmol/mg prot vs (22.52?4.36)nmol/mg prot, P

Objective To investigate the effects of sericin pretreatment on the expression of extracellular matrix(ECM) associated protein in diabetic nephropathy(DN) rats' kidney.Methods Sixty six male SD rats were randomly divided into 3 groups(n=12):normal control group,DN model group and sericine pretreatment group.DN rats model in model group and sericine pretreatment group were established by intraperitoneally injection of streptozotocin(STZ).Blood glucose≥16.7 mmol/L was taken as the standard of successful modelization.The rats in sericine pretreatment group were lavaged with sericine(2.4 g·kg~(-1)·d~(-1)) for 35 days before injecting STZ.The enzymic method was used to measure the blood glucose.Type Ⅳ collagen(cⅣ)and laminin(LN)content in the serum were detected by ELBA.The expression of transforming growth factor-β_1,(TGF-β_1)and tissue inhibitors of maprix metalloproteinase-1(TMP-1) protein in the kidney was observed by immunohistochemical staining.The expression of Smad 3 protein in the kidney was detected by Western blot.Results Compared with normal control rats,the blood glucose,cⅣ and LN content in the serum,TGF-β_1,TIMP-1 and Smad 3 expression in the kidney of the model group rats increased obviously(P<0.01).The blood glucose,cⅣ and LN content in the serum,TGF-β_1,TMP-1 and Smad3 expression in the kidney of rats in sericine pretreatment group were significantly lower than those of the rats in model group(P<0.01).Conclusion Sericin pretreatment can inhibit the activation of TGF-β/Smad 3 signal pathway in the kidney of DN rats,and prevent the decrease of MMPs activity induced by up-regulation of TIMP-1.So sericin can prevent accumulation of ECM and glomerulosclerosis during DN,and has satisfactory apotropaic effects on the development of DN.

Purpose To study clinical pathological significance of the expression of PTEN, p53 and epidermal growth factor receptor ( EGFR) in breast carcinoma and their correlation. Methods Immunohistochemical MaxVision method was used to detect the expres-sion of PTEN, p53, EGFR in 209 cases of infiltrating ductal carcinoma of breast and 40 cases of benign breast diseases. Results The over-expression rate of PTEN, p53 and EGFR protein in breast carcinoma was 57. 9%, 55. 0% and 38. 3% respectively, which was significantly different from those in benign breast diseases (P<0. 05). The expression of PTEN, p53 and EGFR in breast cancer was correlated with tumor size, histological grade, lymph node metastasis, ER status, PR status and molecular subtype (P<0. 05). There was an association between PTEN or p53 and TNM stage, PTEN or EGFR and HER-2 status. There was a negative correlation in the protein expression of PTEN vs EGFR or PTEN vs p53 (P<0. 01). There was a positive correlation between EGFR and p53 protein ex-pression (P=0. 000). The expression of PTEN or EGFR was correlated with the prognosis of breast cancer, but not independent prog-nostic factors. The survival rate of patients with PTEN- /p53 + /EGFR+ was lower than those with other combined expression of PTEN/p53/EGFR (P=0. 008). Conclusions Low or loss expression of PTEN, p53 mutation and EGFR over-expression may be coordinate-ly involved in the pathogenesis and progression of breast cancer. The combined detection of these markers may play an important role in making treatment and indicating prognosis for breast cancer patients.

Bayes Theorem ; Public Health ; Translational Medical Research

Acupuncture Therapy ; Aged ; Aged, 80 and over ; Female ; Humans ; Male ; Pulmonary Disease, Chronic Obstructive ; physiopathology ; therapy

Objective To investigate the changes of epithelial sodium channel(ENaC) expression and sodium and water transport function in the neonatal rat pulmonary edema induced by hyperoxia.Methods The neonatal rats were randomly divided into the hyperoxia group and the control group.After 1,3,5 and 7 d hyperoxia exposure,the lung tissues were collected to measure the wet-to-dry weight ratio and the expression of α-,β-and γ-ENaC subunits were detected by western blot analysis.Alveolar fluid clearance (AFC) and amiloride-sensitive AFC were measured after 5 d to reveal the effect of hyperoxia on the activity of ENaC.Results The lung water contents significantly increased in the hyperoxia group indicating that pulmonary edemahappened(3 d:(6.37 ±0.64) vs (5.56±0.15),t=3.46,P＜0.01;5 d:(5.86 +0.52) vs (5.11±0.21),t=-3.82,P ＜0.01;7 d:(5.56±0.45) vs (4.80±0.09),t =-4.72,P ＜0.01).AFC increased significantly,but no significant difference was found in amiloride-insensitive AFC between the two groups which indicate that amiloride-sensitive AFC increased significantly (AFC:(20.32 ± 3.33) ％ vs (12.97 ± 2.46) ％,t =-6.16,P ＜ 0.01 ; amiloride-insensitive AFC:(10.42 ± 3.44) ％ vs (8.67 ± 3.13) ％,t =-1.30 P =0.21).The expression of α-,β-and γ-ENaC did not reduced after hyperoxia exposure compared with the control group.Conclusion Although bronchopulmonary dysplasia of early pulmonary edema induced by hyperoxia,dysfunctional transport of Na + may not be a key factor involved in pulmonary edema at the early stage of bronchopulmonary dysplasia.

Objective To investigate the expression and significance of Smad4 and Smad7 in newborn rats with hyperoxia-induced chronic lung disease(CLD).Methods Sixty-four newborn Wistar rats 12 h after birth were divided into high-oxygen group (n =32) and air group (n =32,control group) by random number table method.The high-oxygen group was placed in the oxygen glass tank with continuous infusion of oxygen.And 1,3,7,14 d after experiment,tracheal separated,the chest opened to expose heart and lung,slices were Masson staining,undergo dynamic observation of the pulmonary pathological changes under light microscope.Lung fibrosis score was carried out to determine the degree of pulmonary fibrosis,and immunohistochemical technique was used to detect Smad4 and Smad7 protein expression in lung tissue.The expression levels of Smad4 and Smad7 protein in lung tissue were detected with Western blot.Results Compared with the air group,there was statistically significant difference in pulmonary fibrosis score on day 7 (2.67 ± 0.21 vs 0.58 ± 0.17) and day 14 (4.48 ± 0.24 vs 0.63 ± 0.13) in high-oxygen group (P ＜ 0.05) ; Smad4 and Smad7 was main in visible lung epithelial cells and interstitial fibroblasts.Smad4 expression in the high-oxygen group gradually enhanced,compared with the air group (P ＜ 0.05) on day 7 (122.35 ± 10.3 vs 140.08 ±7.77) and day 14(129.7 ± 7.33 vs 144.99 ± 6.49).Smad7 expression in the high-oxygen group first increased and then decreased,expression in the high-oxygen group increased on day 7 (122.35 ± 10.29 vs 130.56 ±9.8),and compared decreased with the air group(P ＜0.05) on day 14(132.16 ±4.38 vs 126.22 ±6.49).Conclusion The newborn rat exposed hyperoxia,the up-regulation of Smad4 protein expression and the down-regulation of Smad7 protein expression are imposible closely related to the happen and development of CLD pulmonary fibrosis.