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.

To study the function and mechanism of the NgR-Rhoa- Rock signal pathways which exists in the retinal ganglion cells apoptosis in diabetes mellitus (DM) rats.
● METHODS: Some healthy SD rats were operated by means of single intraperitoneal injection of 1%streptozotocin based on the standard of 50mg/ kg wight, after that the blood sugar value was greater than 16. 7mmol/ L as DM model, then randomly divided into 3 groups, each group was 10 rats. ln addition to take 10 healthy SD rats as control group. Four groups of rats were bilaterally eyeball intravitreal injection in turn with NgR-siRNA virus 10μ L (siRNA group), NgR-siRNA virus diluted 10μ L ( DM group), NgR - siRNA virus - negative -control solution 10μ L (siRNA blank group), NgR- siRNA virus diluted 10μ L ( normal control group ), and fed normally. During that time, some life indexes like blood glucose, body mass, etc. were measured and recorded. After 12wk, the expression of NgR and Rhoa, HE staining, and TUNNEL staining were detected by Western blot analysis.
● RESULTS: Western blot analysis: compared with normal control group, the expression of NgR and Rhoa in DM group and siRNA blank group increased significantly (P<0. 01), while siRNA group was no significant change (P > 0. 05); compared with DM group and siRNA blank group, the expression of those proteins significantly lowered in siRNA group. HE staining: compared with normal control group, some extent ganglion cells arranged disorder, irregular shape, spacing not consistent were all found in three groups of model rats;compared with DM group and siRNA blank group, there was some improvement in siRNA group of ganglion cells about the order and shape size. TUNEL staining:compared with normal control group, there were retinal ganglion cells apoptosis in all of three groups of model rats. Compared with DM group and siRNA blank group, the number of retinal ganglion cells apoptotic cells was less, and the shape of cells had improved significantly in siRNA group.
●CONCLUSlON: ln the DM phase, the expression of NgR and Rhoa were up - regulation, the condition of diabetic retinal ganglion cell apoptosis was improved after that the NgR-Rhoa-Rock signal pathways had been inhibited.

Objective To investigate the dynamic change of expression of cyclin dependent kinase-4(CDK4)and cyclin dependent kinase inhibitor(p21)in premature rats with hyperoxia-induced chronic lung disease(CLD)and its role.Methods Eighty premature rats were randomly and equally divided into model group(hyperoxia group)and control group(room air group).CLD was induced by hypemxia exposure.The expression of CDK4 and p21 were observed with immunohistochemical method,and the levels of type I collagen were detected by enzyme-linked immunosorbent assay on days 1,3,7,14 and 21.Results Compared with control group,in model group,the expression of CDK4 protein and level of type I collagen statistically increased,but expression of p21 protein decreased significantly on days 14 and 21.The expression of CDK4 was positive correlated with the degree of fibrosis,and expression of p21 was negative correlated with the degree of fibrosis in model group.Conclusion The expression of CDK4 increases and expression of p21 decreases in premature rats exposed to hyperoxia,which may play an important role in the lung fibrosis.

Objective To investigate the renal protection of Chinese cobra venoms (CCV) and its mechanism in renal ischemia/reperfusion (I/R). Methods Thirty-two rats were divided into four groups. 0.1% CCV was separately infused into abdominal cavity at 0.5 h, 24 h before reperfusion in group Ⅰ and Ⅱ . Group Ⅲ suffered from kidney I/R was served as pathological control. Group Ⅳ was sham operation group. BUN and Scr were measured before ischemia and 24 h after reperfusion. Complement C3 was observed at 0, 0.5, 2, 24 h after reperfusion. The kidney samples were examined by HE stain under light microscopy. Apoptosis was detected by terminal deoxynucleotidyl transferase mediated dUTP-biotin in situ nick-end labeling(TUNEL). Results Significant histological damage, apoptosis of tubular cell and impaired renal function were found in group Ⅰ and Ⅲ.The above indexes decreased to a less extend in group Ⅱ (P

Objective To investigate p16 promoter methylation in premature rats with chronic lung disease induced by hyperoxia. Methods Eighty premature Wistar rats were randomly divided into two groups: hyperoxia group (fraction of inspiratory oxygen) 0. 90 and control group (fraction of inspiratory oxygen 0. 21), 40 rats for each group. Semi-nested methylation specific polymerase chain reaction and methylation specific polymerase chain reaction were applied respectively to detect p16 promoter methylation in lung tissues. Additionally, p16 mRNA and protein expressions in lung tissue were detected by reverse transcription- polymerase chain reaction, Western blot and immunohistochemistry method. Results The methylation was not found in control group by seminested methylation specific polymerase chain reaction and methylation specific polymerase chain reaction, while was found in different aged rats of the hyperoxia group. The methylation detection rate was higher by using the semi-nested methylation-specific polymerase chain reaction (52.5%, 21/40) than that by methylation specific polymerase chain reaction (42.5%, 17/40) in the hyperoxia group,but there was no statistically significant difference between the two methods. The p16 mRNA in the hyperoxia group were significantly lower than in the control group at day 7, 14 and 21(1.73 ± 0.40 vs 2.11±0. 37,1.29±0. 19 vs 1.60±0. 27,0. 95±0.25 vs 1.72±0. 34, t=2.19, 2.95 and 10. 43,P＜0. 05). The p16 protein expressions by western blot in the hyperoxia group were significantly lower than in the control group at day 7, 14 and 21 also (88. 1±8. 7 vs 95.0±4.1,65.7±4.5 vs 83. 5±13.6 and 50.4±4.9 vs 86.7±11.9, t=2.27,3.95 and 13.40,P＜0.05). The expression of p16 mRNA (1.06±0.61) and protein (62.32±25.65) in lung tissues of rats with methylation was lower than that without methylation (1.63±0.62 and 94.93±22.21, respectively) (t=2.95, OR=0. 86;t=4.28, OR=0. 85,P＜0.01, respectively). Conclusions Exposure to hyperoxia might induce p16 promoter methylation in lung tissues in premature rats. Methylation risk increases as exposure time extends. p16 promoter methylation induced by hyperoxia might participate in the mechanism of lowering p16 mRNA and protein expression, but might not result in p16 gene silence.