Objective To investigate corneal biomechanical differences in different geometric design parameters of orthokeratology lenses(OK lenses)and to further reveal the corneal shaping mechanism of OK lenses.Methods A coupled finite element model of the aspheric OK lens corneosclera was established for various geometric design parameters of the OK lens,corresponding to different degrees of myopia correction.The distribution trends of the profile and curvature of the corneal anterior surface,as well as the von Mises stress(VMS)and eye axis displacement on both the corneal anterior surface and superior corneal stroma surface were analyzed numerically.Results The stress concentration of the corneal anterior surface was observed in the mid-peripheral and peripheral zones,whereas that of the superior corneal stroma surface appeared in the mid-peripheral zones.The sagittal height of the base curve of the OK lens decreased with increasing degree of myopia correction.At myopia correction degrees of-2.0,-3.0,-4.0,-5.0,and-6.0 D,the maximum corneal VMS increased by 0.81%,1.86%,2.84%,3.81%,and 7.04%,respectively,compared with that at-1.0 D;the curvature of the corneal central zone was reduced by an average of 2.59,3.78,4.51,4.99,5.33,and 6.41 D compared with that without OK lenses.Conclusions The sagittal height of the base curve of the OK lens decreased with increasing degree of myopia correction,resulting in a flatter central curvature of the cornea.The base curve of the OK lens plays a crucial role in both correction and control of myopia.

Objective To investigate the relationship between the therapeutic effect of anti-vascular endothelial growth factor(VEGF)and cytokines in aqueous humor in patients with diabetic retinopathy(DR).Methods From July 2023 to December 2023,50 DR patients(56 eyes)who were treated with anti-VEGF drugs in the Department of Ophthal-mology,the Third Affiliated Hospital of Xinxiang Medical University were included in this study.The patients were divided into the diabetic macular edema(DME)group(30 patients,36 eyes)and the proliferative DR(PDR)group(20 patients,20 eyes).Patients in the DME group received an intravitreal injection of aflibercept once a month for three consecutive times,with aqueous humor extracted before each injection.Patients in the PDR group received an intravitreal injection of aflibercept one week before vitrectomy,with aqueous humor extracted before injection and during vitrectomy,respective-ly.The concentrations of vascular endothelial growth factor A(VEGF-A),placental growth factor(PLGF),interleukin(IL)-1β,IL-23,IL-17A,and tumor necrosis factor-α(TNF-α)in the aqueous humor were detected using the Luminex as-say.Before injection therapy,all patients underwent best corrected visual acuity(BCVA)and central macular thickness(CMT)examinations,and their correlations with cytokine concentrations in DR patients before injection therapy were ana-lyzed.Results Compared to before injection therapy,the concentrations of VEGF-A,PLGF,and IL-23 in the aqueous humor of patients in the DME group decreased significantly after treatment(all P＜0.05);additionally,the CMT de-creased,and the BCVA increased(both P＜0.05).After injection therapy,the concentrations of VEGF,PLGF,IL-1β,IL-23,IL-17A,and TNF-α in the aqueous humor of patients in the PDR group significantly decreased compared to before injec-tion therapy(all P＜0.05).Before injection therapy,the levels of VEGF-A,PLGF,and IL-23 in the aqueous humor of pa-tients in the DME group were higher than those in the PDR group,and the differences were statistically significant(all P＜0.05).The correlation analysis results showed that before injection therapy,VEGF was negatively correlated with BCVA(r=-0.767,P=0.004)and was positively correlated with CMT(r=0.662,P=0.019)and IL-23(r=0.765,P=0.004)in the DME group.There was no correlation between the cytokines in the aqueous humor of patients in the PDR group be-fore injection therapy(all P＞0.05).Conclusion Changes in the concentration of VEGF-A,PLGF,and IL-23 are closely related to the occurrence and development of DR.Anti-VEGF treatment can improve BCVA and decrease CMT in DME pa-tients.The expression level of IL-23 in aqueous humor can serve as a predictive factor for the anti-VEGF efficacy in DR pa-tients,providing new targets for early diagnosis and treatment of DR.

Abstract: Objective To compare the differences in the expression of interleukin-8 (IL-8) and signal transducer and activator of transcription 3 (STAT3) among three groups of individuals: patients with multi-drug resistant tuberculosis (MDR-TB), patients with drug-susceptible tuberculosis (DSP-TB), and healthy controls. By analyzing the differences between these groups, the study seeks to provide a reference basis for clinical auxiliary diagnosis of new type tuberculosis. Methods The differences in mRNA expression profiles of chip from peripheral blood mononuclear cells (PBMCs) among three groups were analyzed. MDR-TB and DSP-TB patients were selected as study subjects, with healthy individuals serving as a control group, and the serum IL-8 concentration of each group was measured using an ELISA assay to compare differences between groups. Additionally, PBMCs from all groups were isolated and total RNA was extracted for analysis of IL-8 and STAT3 mRNA expression using quantitative real-time PCR(Q-PCR). Differences in STAT3 protein expression among the groups were also compared by Western blot assay. Results The serum IL-8 concentration in the MDR-TB and DSP-TB groups was significantly higher than that in the healthy control group, with a statistically significant difference (P<0.05). The relative expression level of IL-8 mRNA in PBMCs of the MDR-TB group and DSP-TB group was also significantly higher than that of the healthy control group, with a statistical significance (P<0.05). Both Q-PCR and Western Blot results revealed that the expression of STAT3 in the healthy, DSP-TB and MDR-TB groups sequentially increased, presenting statistically significant differences between the groups (P<0.05), consistent with the mRNA chip results. Conclusions The expression of IL-8 in active tuberculosis patients (ATB) is significantly increased, which can be used as a marker to distinguish ATB from healthy populations. The changes of STAT3 concentration not only serve as a reference for the diagnosis of ATB but can also distinguish between DSP-TB and MDR-TB populations. The combined detection of IL-8 and STAT3 holds promise as a clinical auxiliary diagnostic basis for different tuberculosis patients.

Objective To investigate biomechanical differences of two posterior occipitocervical internal fixation techniques for treating basilar invagination with atlantoaxial dislocation (BI-AAD). Methods Intra-articular cage + posterior occipital plate+C2 pedicle screw (Cage+C2PS+OP), and intra-articular cage+C1 lateral mass screw+C2PS (Cage+C1LMS+C2PS) models were established based on occipitocervical CT data of the BI-AAD and clinical operation scheme, and the stability of atlantoaxial joint and stress distribution characteristics of C2 endplate and implanted instruments under different motion states were analyzed. Results Compared with the Cage+C1LMS+C2PS model, the atlantoaxial range of motion ( ROM) under flexion, extension, lateral bending and axial rotation in the Cage+C2PS+OP model were reduced by 5. 26% , 33. 33% , 43. 75% , -5. 56% , and stress peak of screw-rod fixation system were reduced by 47. 81% , 60. 90% , 48. 45% , 39. 14% , respectively. Under two internal fixation modes, stresses of C2 endplate and cage were mainly distributed on the compressive side during the motion, and both the screw-bone interface and the caudal side of screw subjected to large loading. Conclusions Two internal fixation methods could provide similar stability. However, the stress concentration of screw-rod system was more obvious and the possibility of screw loosening and fracture was greater under Cage+ C1LMS+C2PS fixation.

Objective To establish a mathematical model of tumor growth and invasion under radiotherapy, so as to numerically simulate the effect of radiotherapy on tumor growth and make sensitivity analysis.Methods The mathematical model of tumor growth and invasion with time evolution before and after radiotherapy was established. The model included four key variables in the process of tumor invasion: tumor cells, extracellular matrix (ECM), matrix-degradative enzymes (MDEs) and oxygen. The linear quadratic (LQ) model was used to simulate the survival probability of tumor cells after radiotherapy, and the effects of different radiotherapy schemes and radiotherapy coefficients on the treatment effect were discussed. Traditional radiotherapy and intraoperative targeted radiotherapy were compared.Results Under the premise of constant total dose, the results of radiotherapy were directly proportional to the radiotherapy coefficient, but not related to the radiotherapy frequency; the therapeutic effect of intraoperative targeted radiotherapy was better than that of standard treatment.Conclusions Simulation results are basically consistent with clinical experimental results. As a more efficient treatment method, intraoperative targeted radiotherapy can provide new ideas for clinical tumor treatment.

The aim of this study was to develop a semi-quantitative immunochromatographic method for rapid detection of Newcastle disease virus (NDV) antibodies by expressing HN protein in rice endosperm bioreactor. The recombinant plasmid pUC57-HN was digested by MlyⅠ and XhoⅠ to retrieve the HN gene, while the intermediate vector pMP3 containing promoter, signal peptide and terminator was digested by NaeⅠ and XhoⅠ. The HN gene and the linearized pMP3 were purified and ligated to form a recombinant plasmid pMP3-HN1. Subsequently, pMP3-HN1 and plant vector pCAMBIA1300 were digested by EcoRⅠ and Hind Ⅲ, and the HN1 gene was cloned into pCAMBIA1300. The recombinant plasmid pCAMBIA1300-HN1 was introduced into Agrobacterium tumefaciens EHA105 by electrotransformation, and the pCAMBIA1300-HN1 was transferred into rice callus by agrobacterium-mediated method. After dark culture, callus screening, differentiation, rooting and transplanting, transgenic rice seeds were obtained 4 months later. PCR identified that the HN gene has been inserted into the rice genome. SDS-PAGE and Western blotting indicated that the HN protein was successfully expressed in the positive rice endosperm. The purity of the HN protein was more than 90% by SP cation exchange chromatography and gel filtration chromatography. According to the national standards for the diagnostic techniques of Newcastle disease HI test (HI≥4log₂, positive antibody reaction), a colloidal gold labeled purified HN protein was used to prepare a semi-quantitative test strip by double-antibody sandwich method for rapid detection of NDV antibody. The results showed that the test strip did not cross-react with positive sera against other viruses, and the sensitivity of the test strip reached 1:102 400 for standard positive sera of Newcastle disease. Testing of a total of 308 clinical sera showed that the compliance rate of the test strip with HI test was 97.08%, and the Kappa value was 0.942. In conclusion, high purity recombinant HN protein was obtained from rice endosperm, and a simple, rapid, highly sensitive and highly specific semi-quantitative immunochromatographic strip was developed. The test strip could be used for immune evaluation of the Newcastle disease vaccine.
Animals ; Antibodies, Viral ; Chickens ; HN Protein/metabolism* ; Newcastle Disease/prevention & control* ; Newcastle disease virus/metabolism* ; Oryza/genetics*

To rapidly and accurately manipulate genome such as gene deletion, insertion and site mutation, the whole genome of a very virulent strain Md5 of Marek's disease virus (MDV) was inserted into bacterial artificial chromosome (BAC) through homogeneous recombination. The recombinant DNA was electroporated into DH10B competent cells and identified by PCR and restriction fragment length polymorphism analysis. An infectious clone of Md5BAC was obtained following transfection into chicken embryo fibroblast (CEF) cells. Furthermore, a lorf10 deletion mutant was constructed by two step Red-mediated homologous recombination. To confirm the specific role of gene deletion, the lorf10 was reinserted into the original site of MDV genome to make a revertant strain. All the constructs were rescued by transfection into CEF cells, respectively. The successful packaging of recombinant viruses was confirmed by indirect immunofluorescence assay. The results of growth kinetics assay and plaques area measurement showed that the lorf10 is dispensable for MDV propagation in vitro. Overall, this study successfully constructed an infectious BAC clone of MDV and demonstrated its application in genome manipulation; the knowledge gained from our study could be further applied to other hepesviruses.
Animals ; Chick Embryo ; Chickens ; Chromosomes, Artificial, Bacterial ; DNA, Recombinant ; Herpesvirus 2, Gallid/genetics* ; Marek Disease

Background: Porcine parvovirus (PPV) is a single-stranded DNA virus that causes porcine reproductive failure. It is of critical importance to study PPV pathogenesis for the prevention and control of the disease. NS1, a PPV non-structural protein, is participated in viral DNA replication, transcriptional regulation, and cytotoxicity. Our previous research showed that PPV can activate nuclear factor kappa B (NF-κB) signaling pathway and then up-regulate the expression of interleukin (IL)-6. Objectives: Herein, the purpose of this study is to determine whether the non-structural protein NS1 of PPV also has the same function. Methods: Real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR), enzyme-linked immunosorbent assay, western blot, immunofluorescence assay and small interfering RNA (siRNA) were used. Results: Our findings demonstrated that PPV NS1 protein can up-regulate the expression levels of IL-6 and tumor necrosis factor-alpha in a dose-dependent manner. Moreover, PPV NS1 protein was found to induce the phosphorylation of IκBα, then leading to the phosphorylation and nuclear translocation of NF-κB. In addition, the NS1 protein activated the upstream pathways of NF-κB. Meanwhile, TLR2-siRNA assay showed TLR2 plays an important role in the activation of NF-κB signaling pathway induced by PPV-NS1. Conclusions These findings indicated that PPV NS1 protein induced the up-regulated of IL-6 expression through activating the TLR2 and NF-κB signaling pathways. In conclusion, these findings provide a new avenue to study the innate immune mechanism of PPV infection.

To establish a novel colloidal gold immunochromatography assay (GICA) for rapid, sensitive and accurate detection of Haemophilus influenzae infection by using the outer membrane protein P6 as detection target. First, the linear antigen epitope located in the extracellular domain of the P6 protein (GenBank accession number: AGH02799) was predicted by bioinformatics analysis. The region (62-75 aa of the protein) with strong antigen specificity was chosen and synthesized. Two rabbits were then immunized by the polypeptides (14 aa) for production of polyclonal antibodies. Then, the recombinant P6 proteins were also obtained to produce polyclonal antibodies. Finally, based on the two antibodies, a novel colloidal GICA for detection of Haemophilus influenzae infection was established and the specificity, sensitivity, repeatability and stability of this method were evaluated. At the same time, the method was tested in clinical simulation, and the plate culture method was used to verify its accuracy. The test strip for Haemophilus influenzae infection was successfully prepared. The detection limit of the test strip was as low as 1×105 CFU/mL and the whole process can be completed within 15 minutes. The strip specifically recognized Haemophilus influenzae and did not react with nine of other common respiratory pathogens such as Streptococcus pneumoniae, Moraxella catarrhalis, Mycoplasma pneumonia, and Legionella pneumophila. And the strips could be stored at 25 °C for at least 6 months without losing sensitivity or specificity. The coincidence rate between the results of 200 clinical samples and the plate culture method was 90.5%. Haemophilus influenzae protein P6, which possessed a high degree of surface antigen accessibility and antigencity, could be used as a marker for Haemophilus influenzae detection. The immunochromatographic colloidal gold test strip which bears the features of rapidity, convenience and sensitivity provides a unique tool for the on-site surveillance and diagnosis of Haemophilus influenzae infection in clinical test.
Animals ; Chromatography, Affinity ; instrumentation ; Diagnostic Tests, Routine ; standards ; Gold Colloid ; chemistry ; Haemophilus Infections ; diagnosis ; Haemophilus influenzae ; Humans ; Limit of Detection ; Rabbits ; Sensitivity and Specificity

A simple and rapid immunochromatographic test strip incorporating a colloidal gold-labeled recombinant Nsp7 antigen probe was successfully developed for the detection of anti-porcine reproductive and respiratory syndrome virus (PRRSV) antibodies in swine. Recombinant Nsp7 protein of PRRSV labeled with colloidal gold was dispensed on a conjugate pad for use as the detector. Staphylococcal protein A and purified porcine anti-Nsp7 antibodies were blotted on a nitrocellulose membrane to form test and control lines, respectively. A comparison of the strip with standard diagnostic tests, enzyme-linked immunosorbent assays and immunoperoxidase monolayer assay, was also performed. The immunochromatographic test strip was shown to be of high specificity and sensitivity. Furthermore, the strip assay is rapid and easy to perform with no requirement for professional-level skills or equipment. It is suggested that the immunochromatographic test strip can be used to quickly and accurately detect PRRSV antibody and to be suitable for diagnostic purposes in the field.
Antibodies* ; Collodion ; Colloids ; Diagnostic Tests, Routine ; Enzyme-Linked Immunosorbent Assay ; Gold Colloid ; Immunochromatography ; Membranes ; Porcine Reproductive and Respiratory Syndrome* ; Porcine respiratory and reproductive syndrome virus* ; Sensitivity and Specificity ; Staphylococcal Protein A ; Swine