Objective To investigate the effect and mechanism of solute carrier family 39 member 14(SLC39A14)on iron overload-induced insulin resistance in HepG2 cells.Methods HepG2 cells were cultured in vitro,and different concentrations of ferric ammonium citrate(FAC)were added to detect cell viability by CCK-8,calcein-AM was used to detect unstable iron pools(LIP),mi-tochondria were observed by transmission electron microscopy,and insulin resistance was detected by cellular glucose consumption,as well as the expression of SLC39A14 protein and mRNA.Subsequently,Ferrostatin-1(Fer-1)was added to detect reactive oxygen spe-cies(ROS),malondialdehyde(MDA)and glutathione(GSH).Finally,small interfering RNA(siRNA)technology was used to knock down the expression of SLC39A14 to observe whether oxidative stress and insulin resistance indices changed.Results After FAC inter-vention,cell viability decreased,the LIP level increased,and glucose consumption reduced.Additionally,the protein and mRNA expres-sion of SLC39A14 was upregulated.Compared with the FAC group,the co-treatment of FAC and Ferrostatin-1 ameliorated the intracel-lular oxidative stress by decreasing the levels of ROS and MDA and increasing the levels of GSH,down-regulating the protein and mRNA expression of SLC39A14,and increasing the glucose consumption,thereby alleviating the onset of insulin resistance.After knockdown of SLC39A14,the levels of oxidative stress and insulin resistance in HepG2 cells in the FAC+SLC39A14 knockdown group were further im-proved compared with those in the FAC group.Conclusion FAC intervention caused the development of insulin resistance in HepG2 cells by a mechanism that may be related to activation of SLC39A14.

BACKGROUND: Visual input significantly influences cerebral activity related to locomotor navigation, although the underlying mechanism remains unclear. This study aimed to analyze the effects of chronic visual impairment and its rehabilitation on sensorimotor integration during level walking in patients with age-related cataract. METHODS: This prospective case series enrolled 14 female patients (68.4 ± 4.7 years) with age-related cataract, scheduled for consecutive cataract surgeries at the Department of Ophthalmology in Peking University Third Hospital from June 2019 to June 2020. Electroencephalogram (EEG) signals during level walking were recorded using a portable EEG system before and 4 weeks after visual restoration. Walking speed was assessed using the Footscan system. Spectral power of the theta and alpha bands was analyzed with repeated-measures analysis of variance, with Assignment (rest and walking), Phase (preoperative and postoperative), and Electrode sites (F3, Fz, F4, O1, and O2) as within-subject factors. RESULTS: Compared to the visual impairment state, theta band power significantly decreased after visual restoration (13.16 ± 1.58 μV 2vs. 23.65 ± 3.48 μV 2 , P = 0.018). Theta activity was notably reduced during walking (17.24 ± 2.43 μV 2vs. 37.86 ± 6.62 μV 2 , P = 0.017), while theta power at rest was not significantly different between the two phases (9.44 ± 1.24 μV 2vs. 9.08 ± 1.74 μV 2 , P = 0.864). Changes in walking speed were correlated with alterations in theta power at electrode sites of O1 ( r = -0.574, P = 0.032) and O2 ( r = -0.648, P = 0.012). Alpha band power remained stable during walking and was unaffected by visual status. CONCLUSIONS Chronic visual impairment from age-related cataract triggers enhanced cerebral activation of sensorimotor integration to compensate for visual decline during locomotion. This cerebral over-activation is effectively alleviated by visual restoration.
Humans ; Female ; Walking/physiology* ; Aged ; Electroencephalography/methods* ; Prospective Studies ; Middle Aged ; Cataract/physiopathology* ; Vision Disorders/physiopathology*

Objective:To discuss whether safranal affects the proliferation,migration,and phenotypic transformation of the vascular smooth muscle cells(VSMCs)in a high-glucose environment and to clarify the function of safranal in the prevention and treatment of diabetic(DM)vascular complications.Methods:The SD rats were selected as experimental subjects;primary VSMCs were cultured from rat thoracic aortas and divided into control group,25 mmol·L-1 high glucose(HG)group,HG+20 μmol·L-1 safranal group,HG+40 μmol·L-1 safranal group,and HG+80 μmol·L-1 safranal group.The cells in control group received no treatment;the cells in 25 mmol·L-1 HG group were pretreated with 25 mmol·L-1 HG;the cells in HG+20,40,and 80 μmol·L-1 safranal groups were further treated with 20,40,and 80 μmol·L-1 safranal respectively for 48 h on the basis of 25 mmol·L-1 HG group.Cell counting kit-8(CCK-8)method was used to determine the appropriate concentration of safranal and detect the viabilities of the VSMCs in various groups;cell scratch healing assay was used to detect the scratch healing rates of the VSMCs in various groups;Transwell chamber assay was used to detect the numbers of the migration VSMCs in various groups;immunofluorescence method was used to detect the fluorescence intensities of alpha-smooth muscle actin(α-SMA)and rabbit anti-osteopontin(OPN)in the VSMCs in various groups;Western blotting method was used to detect the expression levels of OPN,α-SMA,and proliferating cell nuclear antigen(PCNA)in the VSMCs in various groups.Results:Under microscope,on the 4th day of in vitro culture,the spindle-shaped or triangular cells crawled out from the edge of the thoracic aorta tissue blocks,with long spindle being the most common morphology.On the 14th,the cells gradually covered the bottom of the dish;when cell density reached 80%-90%,the characteristic"hills and valleys"growth pattern appeared.Third-generation cells were taken for immunofluorescence identification;immunofluorescence staining with VSMC-specific marker α-SMA showed positive expression of α-SMA protein in the primarily cultured VSMCs.The CCK-8 assay results showed that compared with control group,the cell viability of the cells in 160 μmol·L-1 safranal group was significantly decreased(P<0.01),indicating toxic damage to the cells.Under the conditions of safranal concentrations at 20,40,and 80 μmol·L-1 respectively,after 48 h intervention on VSMCs,no significant adverse effect on cell viability was observed;considering both the effect and toxicity of safranal,these three concentrations were used in subsequent cell experiments.After 48 h intervention,compared with control group,the activity of the VSMCs in 25 mmol·L-1 HG group was increased(P<0.001);compared with 25 mmol·L-1 HG group,the activities of the VSMCs in HG+20,40,and 80 μmol·L-1 safranal groups were gradually decreased(P<0.05).The cell scratch healing assay and Transwell assay results showed that after 48 h intervention,the scratch healing rate of the VSMCs in 25 mmol·L-1 HG group was significantly higher than that in control group(P<0.01),and the number of transmembrane cells through the Transwell chamber was significantly increased(P<0.05);compared with 25 mmol·L-1 HG group,the scratch healing rates of the VSMCs in HG+20,40,and 80 μmol·L-1 safranal groups were gradually decreased(P<0.05),and the number of transmembrane cells was decreased(P<0.05).The immunofluorescence staining results showed that compared with control group,the fluorescence intensity of α-SMA protein in the VSMCs in 25 mmol·L-1 HG group was significantly weakened(P<0.001),while the fluorescence intensity of OPN protein was significantly enhanced(P<0.001);compared with 25 mmol·L-1 HG group,the fluorescence intensities of α-SMA protein in the VSMCs in HG+20,40,and 80 μmol·L-1 safranal groups were gradually increased(P<0.05),and the fluorescence intensities of OPN were gradually weakened(P<0.05).The Western blotting method results showed that compared with control group,the expression level of α-SMA protein in the VSMCs in 25 mmol·L-1 HG group was decreased(P<0.05),and the expression levels of PCNA and OPN proteins were increased(P<0.01);compared with 25 mmol·L-1 HG group,the expression level of α-SMA protein in the VSMCs in HG+20,40,and 80 μmol·L-1 safranal groups were increased(P<0.05),and the expression levels of PCNA and OPN proteins were decreased(P<0.05).Conclusion:Safranal can inhibit the proliferation,migration,and phenotypic transformation of the VSMCs induced by high glucose.

Objective To investigate the effect and mechanism of solute carrier family 39 member 14(SLC39A14)on iron overload-induced insulin resistance in HepG2 cells.Methods HepG2 cells were cultured in vitro,and different concentrations of ferric ammonium citrate(FAC)were added to detect cell viability by CCK-8,calcein-AM was used to detect unstable iron pools(LIP),mi-tochondria were observed by transmission electron microscopy,and insulin resistance was detected by cellular glucose consumption,as well as the expression of SLC39A14 protein and mRNA.Subsequently,Ferrostatin-1(Fer-1)was added to detect reactive oxygen spe-cies(ROS),malondialdehyde(MDA)and glutathione(GSH).Finally,small interfering RNA(siRNA)technology was used to knock down the expression of SLC39A14 to observe whether oxidative stress and insulin resistance indices changed.Results After FAC inter-vention,cell viability decreased,the LIP level increased,and glucose consumption reduced.Additionally,the protein and mRNA expres-sion of SLC39A14 was upregulated.Compared with the FAC group,the co-treatment of FAC and Ferrostatin-1 ameliorated the intracel-lular oxidative stress by decreasing the levels of ROS and MDA and increasing the levels of GSH,down-regulating the protein and mRNA expression of SLC39A14,and increasing the glucose consumption,thereby alleviating the onset of insulin resistance.After knockdown of SLC39A14,the levels of oxidative stress and insulin resistance in HepG2 cells in the FAC+SLC39A14 knockdown group were further im-proved compared with those in the FAC group.Conclusion FAC intervention caused the development of insulin resistance in HepG2 cells by a mechanism that may be related to activation of SLC39A14.

Objective To investigate the prevalence of Cryptosporidium infection and the distribution of parasite species and genotypes among HIV-positive individuals in Jiangxi Province. Methods HIV-positive individuals' sociodemographic and clinical data were collected from three AIDS designated hospitals in Jiangxi Province from January 2022 to March 2023. Subjects' stool samples were collected, and genomic DNA was extracted from stool samples. Nested PCR assay was performed based on the small subunit ribosomal RNA (SSU rRNA) gene of Cryptosporidium, and Cryptosporidium gp60 gene was amplified in stool samples positive for the SSU rRNA gene. The second-round PCR amplification product was checked with 1.5% agarose gel electrophoresis, and the products of suspected positive amplifications were sequenced, followed by sequence alignment. The phylogenetic tree was created using the Neighbor-Joining method with the software MEGA 11.0, to characterize the species, genotypes and sub-genotypes of Cryptosporidium. Results A total of 382 HIV-positive individuals were enrolled, with two cases identified with Cryptosporidium infection (0.52% prevalence), and both cases had no abdominal pain or diarrhea. Following sequencing and sequence alignment, the gene sequences of these two Cryptosporidium isolates shared 99.76% and 99.88% similarity with the gene sequence of C. meleagridis isolates. Phylogenetic analysis based on the Cryptosporidium SSU rRNA gene sequence identified the species of these two Cryptosporidium-positive stool samples as C. meleagridis. Following nested PCR amplification of the Cryptosporidium gp60 gene, sequencing and sequence alignment, the two C. meleagridis isolates were characterized as III eA17G2R1 and III bA25G1R1a sub-genotypes, and the sub-genotype III bA25G1R1a was firstly described in humans. Conclusion The prevalence of Cryptosporidium is low among HIV-positive individuals in Jiangxi Province. The likelihood of Cryptosporidium infection cannot be neglected among HIV-positive individuals without diarrhea.

Objective To investigate the positive rate of HLA-Ⅰ or/and HLA-Ⅱ antibody in female blood donors with a history of pregnancy in Shandong region,with an aim of exploring the significance of HLA antibody testing in reducing clinical platelet transfusion adverse reactions,and providing data support for safe blood transfusion and rational use of blood products.Method Flow cytometry microbead method was used to detect HLA-Ⅰ and HLA-Ⅱ antibodies in 186 samples included in this study.Result Among 186 female blood donors with pregnancy history,77 were detected as positive for HLA-Ⅰ class antibodies(41.4％),41 cases were HLA-Ⅱ positive(22.0％),92 cases were positive for HLA-Ⅰ or/and HLA-Ⅱ antibodies(49.5％).Conclusion A preliminary investigation has conducted on the distribution of HLA antibodies among female blood donors with a history of pregnancy in the local area,and the HLA antibody positivity rate is as high as 49.5％.The results of this study provide a certain data basis for the establishment of preventive measures for transfusion immune related complications caused by blood products and HLA antibodies in a reasonable manner.

Objective To investigate the positive rate of HLA-Ⅰ or/and HLA-Ⅱ antibody in female blood donors with a history of pregnancy in Shandong region,with an aim of exploring the significance of HLA antibody testing in reducing clinical platelet transfusion adverse reactions,and providing data support for safe blood transfusion and rational use of blood products.Method Flow cytometry microbead method was used to detect HLA-Ⅰ and HLA-Ⅱ antibodies in 186 samples included in this study.Result Among 186 female blood donors with pregnancy history,77 were detected as positive for HLA-Ⅰ class antibodies(41.4％),41 cases were HLA-Ⅱ positive(22.0％),92 cases were positive for HLA-Ⅰ or/and HLA-Ⅱ antibodies(49.5％).Conclusion A preliminary investigation has conducted on the distribution of HLA antibodies among female blood donors with a history of pregnancy in the local area,and the HLA antibody positivity rate is as high as 49.5％.The results of this study provide a certain data basis for the establishment of preventive measures for transfusion immune related complications caused by blood products and HLA antibodies in a reasonable manner.

Background::Visual inputs are critical for locomotor navigation and sensorimotor integration in the elderly; however, the mechanism needs to be explored intensively. The present study assessed the gait pattern after cataract surgery to investigate the effects of visual restoration on locomotion.Methods::The prospective study recruited 32 patients (70.1 ± 5.2 years) with bilateral age-related cataracts in the Department of Ophthalmology at Peking University Third Hospital from October 2016 to December 2019. The temporal-spatial gait parameters and kinematic parameters were measured by the Footscan system and inertial measurement units. Paired t-test was employed to compare data normally distributed and Wilcoxon rank-sum test for non-normally distributed. Results::After visual restoration, the walking speed increased by 9.3% (1.19 ± 0.40 m/s vs. 1.09 ± 0.34 m/s, P=0.008) and exhibited an efficient gait pattern with significant decrease in gait cycle (1.02 ± 0.08 s vs. 1.04 ± 0.07 s, P=0.012), stance time (0.66 ± 0.06 s vs. 0.68 ± 0.06 s, P=0.045), and single support time (0.36 ± 0.03 s vs. 0.37 ± 0.02 s, P=0.011). High amplitude of joint motion was detected in the sagittal plane in the left hip (37.6° ± 5.3° vs. 35.5° ± 6.2°, P=0.014), left thigh (38.0° ± 5.2° vs. 36.4° ± 5.8°, P=0.026), left shank (71.9° ± 5.7° vs. 70.1° ± 5.6°, P=0.031), and right knee (59.1° ± 4.8° vs. 56.4° ± 4.8°, P=0.001). The motor symmetry of thigh improved from 8.35 ± 5.30% to 6.30 ± 4.73% ( P=0.042). Conclusions::The accelerated gait in response to visual restoration is characterized by decreased stance time and increased range of joint motion. Training programs for improving muscle strength of lower extremities might be helpful to facilitate the adaptation to these changes in gait.

Here,a styrene-based polymer monolithic column poly(VBS-co-TAT-co-AHM)with reversed-phase/hydrophilic interaction liquid chromatography(RPLC/HILIC)bifunctional separation mode was success-fully prepared for capillary electrochromatography by the in situ polymerization of sodium p-styrene sulfonate(VBS)with cross-linkers 3-(acryloyloxy)-2-hydroxypropyl methacrylate(AHM)and 1,3,5-triacryloylhexahydro-1,3,5-triazine(TAT).The preparation conditions of the monolith were optimized.The morphology and formation of the poly(VBS-co-TAT-co-AHM)monolith were confirmed by scanning electron microscopy(SEM)and Fourier transform infrared spectroscopy(FT-IR).The separation perfor-mances of the monolith were evaluated systematically.It should be noted that the incorporation of VBS functional monomer can provide π-π interactions,hydrophilic interactions,and ion-exchange in-teractions.Hence,the prepared poly(VBS-co-TAT-co-AHM)monolith can achieve efficient separation of thiourea compounds,benzene series,phenol compounds,aniline compounds and sulfonamides in RPLC or HILIC separation mode.The largest theoretical plate number for N,N'-dimethylthiourea reached 1.7×105 plates/m.In addition,the poly(VBS-co-TAT-co-AHM)monolithic column showed excellent reproducibility and stability.This novel monolithic column has great application value and potential in capillary electrochromatography(CEC).

【Objective】 To investigate the serological and molecular genetic characteristics of B(A) and cisAB blood groups discovered in our laboratory. 【Methods】 ABO blood group serology and genetic tests were used to identify blood groups of 6 blood samples, submitted by blood center and hospitals in Shandong, and pedigree investigation was carried on 2 of them. 【Results】 Among the 6 samples, serological results were B(A) in 5 cases and cisAB in 1 case. The results of genetic tests were consistent with the serological results, as the alleles included B(A)04, B(A)02 and cisAB01, and all genotypes were heterozygous with O. Serological pedigree study was conducted on 2 samples: One cisAB patient with his 4 relatives(cisAB type father and three O type relatives) and one B(A)02/O1 donor with his 3 relatives[ B(A) type father/brother and O type mother). For B(A)02/O1 donor, the results of genetic testing were consistent with the serological results, as the paternal genotype was the same as that of the proband, the younger brother was B(A)02/O2, and the maternal genotype was O1/O2. 【Conclusion】 The cisAB and B(A) blood groups are often indistinguishable by serological phenotypes and require genetic confirmation. CisAB pedigree investigation revealed 2 cases of cisAB blood type and B(A) pedigree investigation revealed 3 cases of B(A). The genotyping of cisAB and B(A) in this region were cisAB01/O2, B(A)02/O1, B(A)02/O2, B(A)04/O1 and B(A)04/O2. B(A)and cisAB subtypes can be accurately identified through genetic testing and pedigree investigation, which can provide a reliable basis for blood transfusion selection and ensure the safety of clinical blood transfusion.