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

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.

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.

Objective To explore the role of anterior horn index (AHI) in magnetic resonance imaging (MRI) outcome after intraventricular hemorrhage (IVH) and the correlation factors of IVH in preterm infants.Methods We selected 55 preterm infants with IVH(Ⅱ-Ⅳ degree) confirmed by MRI as case group,and 55 preterm infants with normal MRI as control group,all the cases were admitted in neonatal medical wards in our hospital from January 2009 to September 2011.Then 30 infants of case group were scaned for the second time,to see if they had posthemorrhagic ventricular dilatation (PHVD) and compare the AHI with the last time MRI,and analysis the related factors associated with IVH in preterm infants.Results MRI results:9 infants had PHVD,and the AHI rised (0.36 ± 0.05 v s 0.46 ± 0.06,P ＜ 0.01);21 infants didn't have PHVD,and the AHI decreased (0.31 ± 0.02 vs 0.30 ± 0.02,P ＜ 0.01).Logistic regression analysis:vaginal delivery,use of pulmonary surfactant,mechanical ventrilator care were associated with IVH (P ＜0.05).Conclusion AHI can be used for judging PHVD in preterm infants with IVH.Preterm infants with IVH,especially grade Ⅱ should be monitored AHI.Vaginal delivery and mechanical ventrilator may be risk factor of IVH in preterm infants,while use of pulmonary surfactant may a protective factor of IVH.

Objective To investigate the effect of hyperoxia on fluid transport by fetal distal lung epithelia(FDLE) and the expression of epithelial sodium channel(ENaC) in these cells.Methods FDLE were isolated and randomized into hyperoxia group and normoxia group,which were primarily cultured under hyperoxic or normoxic conditions,respectively.Fluid transport was measured using monolayers of FDLE cultured on Transwell permeable inserts.Western blot was applied to examine the α-ENaC expression.Results Fluid transport across monolayer of FDLE was increased in cells exposed to hyperoxia compaired with cells cultured in normoxic conditions (1.78 ± 0.19 vs 1.06 ± 0.11,P ＜ 0.001).Amiloride significantly decreased the fluid transport in both of the hyperoxia and normoxia groups,but in the presence of amiloride there were no difference between the two groups.The expression of α-ENaC was inhibited by hyperoxia to some extent(24h:0.44 ±0.04 vs 0.40 ±0.04,P=0.22; 48h:0.35 ±0.03 vs 0.47 ±0.06,P =0.03).Conclusion Hyperoxia enhanced total and amiloride-sensitive fluid transport by FDLE.However,the expression of α-ENaC decreased in these cells.

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.

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.