To prepare monoclonal antibody to the N protein of porcine reproductive and respiratory syndrome virus(PRRSV),BALB/c mice were immunized with the purified N protein of the PRRSV-2 strain expressed by prokaryotic expression system.Mouse splenocytes were fused with myeloma cells using hybridoma technique,hybridoma cells were identified by indirect ELISA method and indirect immunofluorescence assay(IFA),and positive hybridoma cells were screened for subclones using the limited dilution method.The results showed that a monoclonal antibody cell line 4A7 was successfully obtained,and the results of Western blot and IFA indicated that the monoclonal antibody could accurately recognize the N proteins of type 1 and type 2 PRRSV.Mean-while,the N protein gene was truncated and expressed by prokaryotic expression system,and the amino acid sequence of the B-cell antigenic epitope recognized by 4A7 was screened as 51EKPHF55 using Western blot.Comparison of epitope amino acids in the N protein gene sequences of different strains revealed that the antigenic epitope 51EKPHF55 recognized by the monoclonal antibody 4A7 has no amino acid difference in the sequences of three subtypes among the PRRSV-1 strains and nine lineages among the PRRSV-2 strains,indicating a high degree of conservation.The results of the study provide a foundation the development of PRRSV diagnostic kits and novel vaccines.

Posterior lenticonus (PLC) is a type of congenital lens disease in the morphology of spherical or conical bulging of the posterior capsule of the lens.It occurs predominantly in unilateral and sporadic, and the bilateral case is often inherited in type of autosomal dominant or X-chromosome linked inheritance.The mutations of EPHA2 are the major known causes of PLC until now.Although researchers have proposed hypotheses that the formation of PLC are related to weak posterior capsule, excessive growth of lens epithelial cells and traction of pigment tissue based on the clinical manifestations of PLC, the pathogenesis of PLC is not elucidated.The animal models associated with PLC can be roughly divided into posterior capsule protrusion and posterior capsule rupture combined with cataract.The clinical manifestations of PLC are various with main symptoms including leukocoria, strabismus, photophobia, and decreased vision.Preoperative examinations by slit-lamp microscopy, ultrasound biomicroscopy, Pentacam can detect abnormal morphological changes of the posterior capsule of lens.Intraoperative characteristic manifestations such as fish-tail and jelly-fish sign can be used to diagnose PLC.When PLC is combined with lens opacity, cataract extraction combined with posterior continuous curvilinear capsulorrhexis or posterior capsule cutting with vitrectomy is the main surgical treatment.Intraocular lens implantation or glass wear is used to correct the aphakic eyes.Postoperative amblyopia training is crucial for improving visual function.

To prepare monoclonal antibody to the N protein of porcine reproductive and respiratory syndrome virus(PRRSV),BALB/c mice were immunized with the purified N protein of the PRRSV-2 strain expressed by prokaryotic expression system.Mouse splenocytes were fused with myeloma cells using hybridoma technique,hybridoma cells were identified by indirect ELISA method and indirect immunofluorescence assay(IFA),and positive hybridoma cells were screened for subclones using the limited dilution method.The results showed that a monoclonal antibody cell line 4A7 was successfully obtained,and the results of Western blot and IFA indicated that the monoclonal antibody could accurately recognize the N proteins of type 1 and type 2 PRRSV.Mean-while,the N protein gene was truncated and expressed by prokaryotic expression system,and the amino acid sequence of the B-cell antigenic epitope recognized by 4A7 was screened as 51EKPHF55 using Western blot.Comparison of epitope amino acids in the N protein gene sequences of different strains revealed that the antigenic epitope 51EKPHF55 recognized by the monoclonal antibody 4A7 has no amino acid difference in the sequences of three subtypes among the PRRSV-1 strains and nine lineages among the PRRSV-2 strains,indicating a high degree of conservation.The results of the study provide a foundation the development of PRRSV diagnostic kits and novel vaccines.

Posterior lenticonus (PLC) is a type of congenital lens disease in the morphology of spherical or conical bulging of the posterior capsule of the lens.It occurs predominantly in unilateral and sporadic, and the bilateral case is often inherited in type of autosomal dominant or X-chromosome linked inheritance.The mutations of EPHA2 are the major known causes of PLC until now.Although researchers have proposed hypotheses that the formation of PLC are related to weak posterior capsule, excessive growth of lens epithelial cells and traction of pigment tissue based on the clinical manifestations of PLC, the pathogenesis of PLC is not elucidated.The animal models associated with PLC can be roughly divided into posterior capsule protrusion and posterior capsule rupture combined with cataract.The clinical manifestations of PLC are various with main symptoms including leukocoria, strabismus, photophobia, and decreased vision.Preoperative examinations by slit-lamp microscopy, ultrasound biomicroscopy, Pentacam can detect abnormal morphological changes of the posterior capsule of lens.Intraoperative characteristic manifestations such as fish-tail and jelly-fish sign can be used to diagnose PLC.When PLC is combined with lens opacity, cataract extraction combined with posterior continuous curvilinear capsulorrhexis or posterior capsule cutting with vitrectomy is the main surgical treatment.Intraocular lens implantation or glass wear is used to correct the aphakic eyes.Postoperative amblyopia training is crucial for improving visual function.

Objectives:To explore the effect and possible mechanisms of mild hypothermia on interferon(IFN)-α2b-induced AC16 cardiomyocytes apoptosis. Methods:Cardiomyocytes were stimulated in ordinary temperature and mild hypothermia by IFN-α2b under different concentrations for different times.Proliferation activity of cardiomyocytes was detected by CCK-8 assay.Apoptosis was detected by flow cytometry technique.The effects of different interventions on mitochondrial morphology were examined using Mito-Tracker Green and laser scanning confocal microscope,respectively.The mitochondrial membrane potentials under different intervention conditions were detected by flow cytometry.The fusion of dynamin-related protein 1(Drp1)and mitochondria,and the effects of different interventions on the mitochondria was examined by Drp1 or mitochondrial fluorescent probes and laser scanning confocal microscope.The effects of different intervention conditions on the protein expression level of Phospho-Drp1(p-Drp1)Ser616,Drp1,cleaved poly ADP-ribose polymerase1(cleaved-PARP1),poly ADP-ribose polymerase1(PARP1)were detected by Western blot. Results:CCK-8 assay and flow cytometry results showed that IFN-α2b inhibited the proliferation and enhanced the apoptosis of AC16 cardiomyocytes in a time and dose-dependent manner,these effects could be attenuated by mild hypothermia.Mito-Tracker Green,laser scanning confocal microscope and flow cytometry results showed that the extent of damage of mitochondria with different interventions were attenuated in the setting of mild hypothermia as compared with ordinary temperature.The morphology of mitochondria remained intact and the mitochondrial membrane potentials were the highest in mild hypothermia group.Injured AC16 cardiomyocytes released Drp1 from cytoplasm to mitochondria and increased mitochondrial fission,these effects were abolished after mild hypothermia.p-Drp1 Ser616/Drp1 ratio and cleaved-PARP1/PARP1 ratio were decreased after mild hypothermia,and above effects could be reversed by mitochondrial division inhibitor-1(Mdivi-1)pretreatment. Conclusions:Mild hypothermia inhibits IFN-α2b-induced AC16 cardiomyocytes apoptosis via improving mitochondrial function.

Objective To prepare a nano drug(PFOB@Lip-MMC)with liposome as the carrier,liquid perfluorooc-tyl bromide(PFOB)as core and mitomycin C(MMC)loading on the liposome shell and study its inhibitory effect on the proliferation of human pterygium fibroblasts(HPFs).Methods The thin film dispersion-hydration ultrasonic method was used to prepare PFOB@Lip-MMC and detect its physical and chemical properties.Cell Counting Kit-8,Cam-PI cell viability staining and flow cytometry were employed to detect the impact of different concentrations of PFOB@Lip-MMC on the via-bility of HPFs.DiI fluorescence labeled PFOB@Lip-MMC was used to observe the permeability of the nano drug to HPFs under a laser confocal microscope.After establishing HPF inflammatory cell models,they were divided into the control group(with sterile phosphate-buffered saline solution added),PFOB@Lip group(with PFOB@Lip added),MMC group(with MMC added),PFOB@Lip-MMC group(with PFOB@Lip-MMC added)and normal group(with fresh culture medi-um added)according to the experimental requirements.After co-incubation for 24 h,flow cytometer was used to detect the apoptosis rate of inflammatory cells,and the gene expression levels of interleukin(IL)-1β,prostaglandin E2(PGE2),tumor necrosis factor(TNF)-α and vascular endothelial growth factor(VEGF)in cells were analyzed by PCR.Results The average particle size and Zeta potential of PFOB@Lip-MMC were(103.45±2.17)nm and(27.34±1.03)mV,respec-tively,and its entrapped efficiency and drug loading rate were(72.85±3.28)％and(34.27±2.04)％,respectively.The sustained-release MMC of drug-loaded nanospheres reached(78.34±2.92)％in vitro in a 24-hour ocular surface environ-ment.The biological safety of PFOB@Lip-MMC significantly improved compared to MMC.In terms of the DiI fluorescence labeled PFOB@Lip-MMC,after co-incubation with inflammatory HPFs for 2 h,DiI fluorescence labeling was diffusely dis-tributed in the cytoplasm of inflammatory HPFs.The apoptosis rate of inflammatory HPFs in the PFOB@Lip-MMC group[(77.23±4.93)％]was significantly higher than that in the MMC group[(51.62±3.28)％].The PCR examination results showed that the gene transcription levels of IL-1 β,PGE2,TNF-α and VEGF in other groups were significantly reduced com-pared to the control group and PFOB@Lip group,with the most significant decrease in the PFOB@Lip-MMC group(all P＜0.05).Conclusion In this study,a novel nano drug(PFOB@LIP-MMC)that inhibited the proliferation of HPFs was successfully synthesized,and its cytotoxicity was significantly reduced compared to the original drugs.It has good bio-compatibility and anti-inflammatory effects,providing a new treatment approach for reducing the recurrence rate after pte-rygium surgery.

Objective:To investigate the effect of multiple medications on the risk of small intestinal bacterial overgrowth(SIBO)in the elderly.Methods:Clinical data of 85 inpatients in the Department of Geriatrics, the First Hospital of Lanzhou University undergone HMBT from August 2017 to April 2021 were retrospectively analyzed.According to the HMBT results, they were divided into a SIBO(+ )group and a SIBO(-)group.Polypharmacy was defined as ≥ 5 types of medications.We analyzed the difference in the rate of polypharmacy between the two groups.Results:A total of 85 hospitalized elderly patients were included in the study.Of these patients, 38(44.71%)tested positive for SIBO.Polypharmacy occurred in 41 patients(48.24%). There were significant differences in types of drugs and polypharmacy between the SIBO(+ )group and the SIBO(-)group( t=3.01 and χ2=14.33, P<0.05 for both). Moreover, polypharmacy was a risk factor for SIBO( P=0.017, OR=10.85, 95% CI: 1.52-77.29). Among 14 commonly used drugs, gastrointestinal motility drugs were closely related to SIBO.There was a positive correlation between polypharmacy and the change in hydrogen levels at 90 min( P=0.040, r=0.22, 95% CI: 0.01-0.42). Conclusions:Polypharmacy is correlated with SIBO in the elderly, is a risk factor for SIBO and is helpful in clinical practice to assess the risk of SIBO and decide further examinations, contributing to early diagnosis and early treatment.