AIM: To develop a novel gene-delivery therapeutic based on CRISPR/Cas9 genome editing technology capable of specifically targeting and knocking out the VEGFA gene, thereby achieving sustained suppression of VEGFA expression in retinal pigment epithelial(RPE)cells and providing a new strategy for gene therapy in retinal neovascular diseases.METHODS:Single guide RNAs targeting the human VEGFA gene for knockout were designed, and corresponding recombinant plasmids were constructed. A novel polymer(PTEE)was used to encapsulate the plasmids to prepare a PTEE-loaded anti-VEGFA plasmid(PLAP)gene delivery system. PTEE materials at concentrations of 0.1, 0.2, 0.4, 0.8, and 1.6 μg/μL were co-incubated with ARPE-19 cells, and the biocompatibility of PTEE was evaluated using the cell counting kit-8(CCK-8)assay. Recombinant plasmids expressing green fluorescent protein(GFP)were constructed. Lipofectamine 3000 and jetOPTIMUS®DNA transfection reagents were used as control groups, and PTEE nanomaterials were used as the experimental group to encapsulate the plasmids. When the cell confluence reached 80%, the formulations were transfected into ARPE-19 and 293T cells. GFP expression was observed under light microscopy, and the transfection efficiencies of each group were compared. ARPE-19 cells were induced under hypoxia, and PLAP was transfected into the cells. The expression level of VEGFA was detected by enzyme-linked immunosorbent assay(ELISA)to evaluate the efficacy of this novel gene delivery system.RESULTS: After co-incubation of ARPE-19 cells with different concentrations of PTEE for 24 h and 48 h, no significant effect on cell viability was observed in any group. The transfection efficiency of PLAP in ARPE-19 cells was higher than that in the Lipo3000 and jetOPTIMUS groups, with statistically significant differences(P<0.01). Hypoxia for 6 h significantly induced the upregulation of VEGFA mRNA expression in ARPE-19 cells, and under hypoxic conditions, the PTEE group exhibited a significant inhibitory effect on VEGFA expression(P<0.01).CONCLUSION:PLAP exhibits favorable biocompatibility and prominent VEGFA inhibitory effects in vitro, making it a potential candidate drug for gene therapy of retinal neovascular diseases.

Diabetes is one of the major chronic non-communicable diseases in China. This article discusses the progress of diabetes prevention and control under the leadership of the government and how to integrate the National Diabetes Prevention and Control Center (DPCC) into the work of primary medical and health institutions in Yunnan Province, providing reference and experience for diabetes prevention and control efforts in the central and western regions of China.

Objective Aimed at investigating the effect and molecular mechanism of artemisinin on hemo-dynamics and vascular remodeling in monocrotaline(MCT)-induced pulmonary arterial hypertension(PAH)rats.Methods 30 male SD rats were randomly divided into 3 groups(n=10):control group,MCT-induced PAH group(MCT group,60 mg/kg)and artemisinin intervention group(50 mg/kg).At 28 days after modeling,the right ventricular systolic pressure(RVSP),mean pulmonary artery pressure(mPAP),heart rate and right ventricular hypertrophy index(RVHI)were measured to evaluate the development of PAH.HE staining and α-SMA immuno-histochemistry were used to observe the morphology and assess muscularization of pulmonary arterioles,and the percentage of medial wall thickness(WT%),the percentage of vascular wall area(WA%)and the proportion of muscular vessels were calculated to evaluate the degree of pulmonary vascular remodeling.The mRNA and protein levels of HIF-1α and LDHA were detected by real-time PCR and Western blot,respectively.Pyruvate and lactate concentration in lung tissue was measured using pyruvate and lactateassay kit.Results Compared with the control group,the RVSP,mPAP,heart rate and RVHI were significantly increased in MCT-induced PAH rats(all P<0.05).Histological analysis showed that the increasedmedial wall thickness of small pulmonary arteries and vascular muscularization were observed in MCT-treated rats compared with control rats.WT%,WA%and muscularization degrees of pulmonary arterioles were higher in MCT-treated rats than those in the control group(all P<0.05),suggesting successful construction of PAH model.Compared with the MCT group,the RVSP,mPAP,heart rate and RVHI decreased in the rats treated with artemisinin(all P<0.05),accompanied with lower WT%and WA%(P<0.05),and muscularization of pulmonary arterioles was improved(P<0.05).Further study showed the mRNA and protein levels ofHIF-1α and LDHA in lung tissue of MCT-induced PAH rats were higher than those in the control group,the content of lactate and pyruvate and the ratio of lactate to pyruvate were higher than that in the control group(all P<0.05).However,the mRNA and protein levels of HIF-1α and LDHA in lung tissue of rats treated with artemisinin were lower than those in the MCT group,the content of lactate and pyruvate and the ratio of lactate to pyruvate were lower than that in the MCT group(all P<0.05).Conclusion Artemisinin improves hemodynamic and pulmonary vascular remodeling in PAH rats through inhibiting HIF-1α/LDHA signaling pathway-mediated glycolysis.

Purpose To evaluate the diagnostic value of three-dimensional amide proton transfer weighted imaging(APTWI)in distinguishing benign from malignant bone tumors and the diagnostic efficacy of APTWI combined with diffusion-weighted imaging(DWI).Materials and Methods Sixty-nine patients with pelvic and lower extremity bone tumors,confirmed by puncture or surgical pathology at the First Affiliated Hospital of Zhengzhou University,were prospectively collected from April 2022 to July 2023.On the relevant parameter maps of APTWI and DWI,the asymmetric magnetization transfer rate at a chemical shift of 3.5 ppm[MTRasym(3.5 ppm)]and apparent diffusion coefficient(ADC)values corresponding to the lesions were measured,respectively.Comparative analysis of differences in each parameter was conducted,and the diagnostic efficacy of these parameters alone and in combination was compared.Results Among 69 bone tumor,24 were benign cases and 45 malignant cases.MTRasym(3.5 ppm)of malignant bone tumors was higher than that of benign tumors[(2.974±0.630)%vs.(2.215±0.963)%],while ADC values of malignant bone tumors were lower than those of benign tumors[(1.143±0.406)×10-3 s/mm2 vs.(1.757±0.449)×10-3 s/mm2],with significant differences between the two groups(t=3.532,-5.645;P<0.001).Receiver operating characteristic curve showed that MTRasym(3.5 ppm),ADC values,and their combination exhibited good performance in the differential diagnosis of benign and malignant bone tumors,with the area under the curve being 0.765,0.841 and 0.874,respectively.There were no statistically significant differences in the area under the curve between the combined APTWI-DWI and either APTWI or DWI alone in diagnosing benign and malignant bone tumors(combined vs.APTWI:Z=1.873,P=0.064;combined vs.DWI:Z=1.333,P=0.167).The diagnostic specificity of combined APTWI-DWI was significantly higher than that of APTWI alone(96%vs.56%,P=0.001),but there was no statistically significant difference compared with DWI alone(96%vs.80%,P=0.189).Conclusion APTWI and DWI,either alone or in combination,could serve as effective imaging indicators for distinguishing benign from malignant bone tumors.The combination of APTWI and DWI shows a trend of achieving higher diagnostic efficacy and diagnostic specificity in differentiating benign from malignant bone tumors,thus holding potential clinical application value.

Promoting the construction of medical union is an important part of deepening the reform of medical and health system,and an effective means to optimize the allocation of medical resources in the region.It will take the construction and development of the compact medical union in Guangdong Province as the research subject,ana-lyze the current common problems in the development of the compact medical union,and take the construction practice of the compact medical union in a large public hospital as the research object,explore and analyze the clas-sification and experience effect of three models of the compact medical union,with a view to putting forward sug-gestions and measures with certain reference significance.

Diabetes is one of the major chronic non-communicable diseases in China. This article discusses the progress of diabetes prevention and control under the leadership of the government and how to integrate the National Diabetes Prevention and Control Center (DPCC) into the work of primary medical and health institutions in Yunnan Province, providing reference and experience for diabetes prevention and control efforts in the central and western regions of China.

To identify the bacterial pathogens in a case of goat respiratory disease in Yunnan prov-ince,three Gram-negative,rod-shaped bacteria strains,YN240861,YN240862 and YN240863,were isolated from lung samples of three diseased goats with respiratory disease,and the identifications of the biochemical and the sequence similarity and phylogeny of 16S rDNA gene,and the analyses of the pathogenicity and antimicrobial susceptibility were studied.The results of the bacterial iden-tification indicated that the most biochemical characteristics of the three isolates were identical with those of the Histophilus somni(H.somni)type stain ATCC 43625T and the other reference strains of H.somni.The three strains had the 16S rDNA sequence similarity of 97.7％with H.som-ni type strain ATCC 43625T,and shared the 16S rDNA sequence similatiies of 99.4％-99.9％with the other H.somni reference strains.The three strains formed the same branch with all the H.som-ni reference strains in the 16S rDNA gene sequence phylogenetic tree.Based on these identification results,the three strains were identified as H.somni.The mouse pathogenicity test showed that each of the three strains had the median lethal dose(LD50)≤7.6×10 5 CFU in mice,which indica-ted that the three stains have higher pathogenicity to mice.The results of the antimicrobial suscep-tibility test indicated that the three strains were sensitive to ampicillin of penicillins,cefazolin and ceftriaxone of cephalosporins,chloramphenicol and florfenicol of chloramphenicols,kanamycin and gentamicin of aminoglycosides,and ofloxacin of quinolones.The strain YN240862 was resistant to sulfamethoxazole of sulfonamides.To our knowledge,this is the first isolation of Hi.somni in Chi-na,and confirmed the presence of H.somni infection in goats in China,which provides the basic da-ta for the prevention and treatment of H.somni infection in goats in China.

Objective Aimed at investigating the effect and molecular mechanism of artemisinin on hemo-dynamics and vascular remodeling in monocrotaline(MCT)-induced pulmonary arterial hypertension(PAH)rats.Methods 30 male SD rats were randomly divided into 3 groups(n=10):control group,MCT-induced PAH group(MCT group,60 mg/kg)and artemisinin intervention group(50 mg/kg).At 28 days after modeling,the right ventricular systolic pressure(RVSP),mean pulmonary artery pressure(mPAP),heart rate and right ventricular hypertrophy index(RVHI)were measured to evaluate the development of PAH.HE staining and α-SMA immuno-histochemistry were used to observe the morphology and assess muscularization of pulmonary arterioles,and the percentage of medial wall thickness(WT%),the percentage of vascular wall area(WA%)and the proportion of muscular vessels were calculated to evaluate the degree of pulmonary vascular remodeling.The mRNA and protein levels of HIF-1α and LDHA were detected by real-time PCR and Western blot,respectively.Pyruvate and lactate concentration in lung tissue was measured using pyruvate and lactateassay kit.Results Compared with the control group,the RVSP,mPAP,heart rate and RVHI were significantly increased in MCT-induced PAH rats(all P<0.05).Histological analysis showed that the increasedmedial wall thickness of small pulmonary arteries and vascular muscularization were observed in MCT-treated rats compared with control rats.WT%,WA%and muscularization degrees of pulmonary arterioles were higher in MCT-treated rats than those in the control group(all P<0.05),suggesting successful construction of PAH model.Compared with the MCT group,the RVSP,mPAP,heart rate and RVHI decreased in the rats treated with artemisinin(all P<0.05),accompanied with lower WT%and WA%(P<0.05),and muscularization of pulmonary arterioles was improved(P<0.05).Further study showed the mRNA and protein levels ofHIF-1α and LDHA in lung tissue of MCT-induced PAH rats were higher than those in the control group,the content of lactate and pyruvate and the ratio of lactate to pyruvate were higher than that in the control group(all P<0.05).However,the mRNA and protein levels of HIF-1α and LDHA in lung tissue of rats treated with artemisinin were lower than those in the MCT group,the content of lactate and pyruvate and the ratio of lactate to pyruvate were lower than that in the MCT group(all P<0.05).Conclusion Artemisinin improves hemodynamic and pulmonary vascular remodeling in PAH rats through inhibiting HIF-1α/LDHA signaling pathway-mediated glycolysis.

BACKGROUND:The highest incidence of spinal fracture is in the thoracolumbar segment,and its symptoms are back pain,posterior convexity deformity,activity limitation,or with spinal cord nerve injury causing lower limb pain,numbness,and even paraplegia and other complications.The finite element method is a digital computer modeling technique,which can simulate the physical model and carry out force analysis realistically.OBJECTIVE:To review the application of finite element method in thoracolumbar spine fractures.METHODS:We searched the Chinese and English literature databases PubMed,Web of Science,and CNKI for relevant literature on the application of the finite element analysis method in spinal thoracolumbar fracture published before March 2024.The search terms in Chinese and English were:finite element analysis methods,biomechanical phenomena,stress analysis,thoracolumbar fractures,spinal fractures.Finally,55 papers were included.RESULTS AND CONCLUSION:(1)The exploration of thoracolumbar fractures caused by different etiologies(osteoporotic,traumatic,and pathological)through the finite element method is conducive to a deeper understanding of the biomechanics of various types of thoracolumbar fractures,and to improve the individualized and fine-tuned treatment of thoracolumbar fractures.(2)The finite element analysis of a single sample or a small number of samples has the chance,and a larger number of samples are required for the future finite element analysis to reduce the chance caused by the sample.(3)The rigid structure of bones alone cannot meet the biomechanical working conditions of the integrity of the physical object,and future finite element models need to incorporate all the structures of the physical object(e.g.,soft tissues,such as muscles and ligaments)as far as possible.(4)The finite element method has been used in more studies on osteoporotic and traumatic thoracolumbar spine fractures,which will need to be more in-depth in the future,and less in the field of pathologic thoracolumbar fractures,which has a wider scope for future research.

Objective To investigate the application value of arterial spin labeling(ASL)perfusion weighted imaging in the early diagnosis of Alzheimer's disease(AD).Methods A total of 60 patients with different ages(control group)and 60 AD patients with different degrees(patient group)who underwent cerebral ASL perfusion weighted imaging examination were selected.At the work-station,the region of interest(ROI)of the hippocampus and limbic system brain structures were delineated and the cerebral blood flow(CBF)value of the ROI were measured.The CBF values of the brain structures of the control group at different ages and the corresponding ROI of the control group and the patient group were compared.Results There were statistically significant differences in the CBF values of the cingulate gyrus,parahippocampal gyrus,hippocampus,orbital part of frontal lobe,temporal pole of temporal lobe,insula lobe and amygdaloid body of the limbic system among the different ages in the control group(P＜0.05),and the CBF value of brain structures gradually decreased with the increase of age.There were statistically significant differences in CBF values of ROI between the control group and mild,moderate,severe patient group(P＜0.05),and the CBF value of brain structures gradually decreased with the severity of AD condition.Conclusion The CBF value of ASL perfusion weighted imaging can objectively reflect the blood flow changes and functional status of the hippocampus and limbic system brain structures in elderly people of different ages and AD patients of different degrees,and has important value in the early diagnosis of AD patients.