Objective:To explore the clinical phenotype, genotype and genetic characteristics for a patient with unilateral paravenous retinochoroidal atrophy (PPRCA) and retinitis pigmentosa (RP) in the contralateral eye.Methods:A PPRCA pedigree which had presented at the Department of Medical Genetics of the Eye Hospital of Wenzhou Medical University in August 2021 was selected as the study subject. Clinical data of the family members were collected. The proband underwent wide-field fundus photography, wide-field autofluorescence, full-field electroretinogram (ff-ERG), visual field testing, optical coherence tomography (OCT), and fundus angiography (FFA and ICGA). Blood samples were collected from the proband and family members (parents and two sisters), and buccal mucosal cells were collected from the proband′s daughter, and genomic DNA was extracted for each family member. Whole exome sequencing (WES) was performed on the proband. Candidate variants were verified using Sanger sequencing and pathogenicity analysis. This study was approved by the Medical Ethics Committee of the Eye Hospital of Wenzhou Medical University (Ethics No.2021-212-K-185).Results:① Wide-angle fundus photography and autofluorescence showed that the right eye of the proband was consistent with PPRCA and the left eye with RP. OCT showed that the outer layer of the fovea was intact in the right eye, while disorganized outer segment was found in the fovea of the left eye, and outer segment atrophies outside the fovea were found in both eyes. The amplitudes of ff-ERG decreased significantly in both eyes, and the amplitudes in right eye were slightly higher than those of the left eye. Visual field showed a paracentral arcuate scotoma in the right eye and severe centripetal contraction in the left eye. FFA showed hyperfluorescence in the retinal vein distribution area caused by atrophy of retinal pigment epithelium of the right eye and hypofluorescence related to bone spicule pigmentation, in addition with mottled hypofluorescence of choroid in the left eye. ICGA showed mild paravenous retinochroidal atrophy of the right eye and diffuse choroid capillaries atrophy in the middle and peripheral area of the left eye. ② WES revealed that the proband had a heterozygous c. 2234C>T (p.Thr745Met) variant of the CRB1 gene. Sanger sequencing confirmed that the proband and family members except the father of the proband carried the same CRB1 gene variant. ③ Based on the "criteria and guidelines for the classification of genetic variation" and related consensus from the American College of Medical Genetics and Genomics (ACMG), this variant was classified as pathogenic (PM3_VeryStrong+ PM1+ PM2_Supporting + PP3). Conclusion:The heterozygous c. 2234C>T (p.Thr745Met) variant of the CRB1 gene may underlay the unilateral PPRCA with contralateral eye RP in this proband. Above findings have enriched the mutational spectrum of the CRB1 gene.

Objective:To investigate the interventional effects of the Fuzheng Huayu (FZHY) formula and its partial mechanistic role on cholestatic liver fibrosis in mice.Methods:Mdr2 gene knockout (Mdr2－/ －) mice were randomly divided into a model group, FZHY group, and Obeticholic acid group. Wild-type C57BL/6J mice of the same age served as the control group. Mdr2－/ －mice were given the corresponding drugs starting from the first day of 9 weeks of age by oral gavage in each group. The control and model groups were administered 0.3% sodium carboxymethylcellulose by oral gavage and were sacrificed at 12 weeks of age for specimen collection. High-speed biochemistry analyzer was used to detect serum alkaline phosphatase and alanine aminotransferase activity in mice. Hematoxylin-eosin staining and Sirius red staining were used to observe pathological changes in liver tissues. Hydroxyproline content was measured to assess collagen in liver tissues. Immunohistochemical staining, Western blotting, and real-time fluorescence quantitative PCR were used to detect the expression of fibrosis markers Col-I and alpha-smooth muscle actin in liver tissues. The expressional condition of cholangiocyte response markers Epcam, CK7, CK19, as well as Pcna, Mki67, and Ccnd1, inflammatory related factors Ccl2, Ccl5, Tnf-α, Il10, and Cxcl4, phosphorylated peroxisome proliferator-activated receptor alpha (PPARα) and nuclear factor kappa-B (NF-κB) were determined. Comparative analysis among multiple groups was performed using one-way ANOVA. The LSD method was used for comparisons between groups. Two-tailed statistical tests were used.Results:Compared with wild-type mice, Mdr2 －/ － mice had a significant increase in serum alanine aminotransferase and alkaline phosphatase activity ( P<0.001). The percentage of Sirius red-positive staining areas and hydroxyproline content in liver tissues was significantly increased ( P<0.01). The expression of Col-I, α-smooth muscle actin, Epcam, CK7, CK19, Pcna, Mki67, and Ccnd1, and the expression of Ccl2, Ccl5, Tnf-α, Il10, and Cxcl4 were significantly increased ( P<0.01); however, both FZHY and Obeticholic acid significantly reversed the increases in these indicators ( P<0.05; P<0.01). Further results showed that compared to wild-type mice, the expression of PPARα was significantly reduced in liver tissues of Mdr2 －/ － mice, while NF-κB was significantly enhanced ( P<0.01). In contrast, compared to Mdr2-/- mice, the expression of PPARα in the liver tissues of FZHY group mice was significantly increased ( P<0.05), while NF-κB was significantly inhibited ( P<0.05). Conclusion:FZHY can significantly improve liver fibrosis, cholangiocyte response, and inflammation in Mdr2 －/ － mice with spontaneously occurring cholestatic liver fibrosis, and its mechanistic role is related to the regulation of the PPARα/NF-κB pathway.

Objective: To explore the effect of interferon regulatory factor 2 binding protein 2 ( IRF2BP2) on the proliferation of acute myeloid leukemia ( AML) cells and its molecular mechanism． Methods: The CRISPR-Cas9 gene editing technology was used to knock out IRF2BP2 in human AML cell lines Kasumi-1 and U937，and West- ern blot was performed to detect the knockout efficiency of IRF2BP2 protein．Cell morphology was observed using a microscope．Cell phenotypes were analyzed by CCK-8 assay，colony formation experiments，and flow cytometry． RNA-Seq was performed to identify differentially expressed genes between the IRF2BP2 knockout group and the control group in the U937 cell line．Gene Set Enrichment Analysis ( GSEA) was conducted to explore the down- stream molecular mechanisms．Western blot was used to detect the expression of downstream differentially expressed genes．The Cleavage Under Targets and Tagmentation ( CUT＆Tag) technique was applied to identify the direct tar- gets of the IRF2BP2 protein，and the corresponding binding signals were visualized using the Integrated Genomics Viewer ( IGV) ． Results: Compared with the control group，after knocking out IRF2BP2，the CCK-8 experiment showed that AML cell proliferation was inhibited ( P ＜0. 05) ; the number of colonies in the IRF2BP2 knockout group decreased ( P＜0. 05) ，and the proportion of G1 phase was prolonged ( P＜0. 05) ; in U937 cell lines，knoc- king out IRF2BP2 resulted in significant enrichment of differential genes in myelocytomatosis oncogene ( MYC) -re- lated signaling pathways，and the protein expression levels of pathway molecules MYC，cyclin-dependent kinase 4 ( CDK4) ，and cyclin － dependent kinase 2 ( CDK2 ) decreased with the downregulation of IRF2BP2; using IRF2BP2 antibodies in U937 cell lines for CUT＆Tag experiments，IGV visualization analysis showed a significant increase in signal peaks in the MYC promoter region． Conclusion IRF2BP2 protein affects the cell cycle and pro- liferation of AML cells by targeting and regulating MYC．

OBJECTIVE: To explore the clinical phenotype, genotype and genetic characteristics for a patient with unilateral Pigmented paravenous retinochoroidal atrophy (PPRCA) and Retinitis pigmentosa (RP) in the contralateral eye. METHODS: A PPRCA pedigree which had presented at the Department of Medical Genetics of the Eye Hospital of Wenzhou Medical University in August 2021 was selected as the study subject. Clinical data of the family members were collected. The proband underwent wide-field fundus photography, wide-field autofluorescence, full-field electroretinogram (ff-ERG), visual field testing, optical coherence tomography (OCT), and fundus angiography (FFA and ICGA). Blood samples were collected from the proband and family members (parents and two sisters), and buccal mucosal cells were collected from the proband's daughter, and genomic DNA was extracted for each family member. Whole exome sequencing (WES) was performed on the proband. Candidate variants were verified using Sanger sequencing and pathogenicity analysis. This study was approved by the Medical Ethics Committee of the Eye Hospital of Wenzhou Medical University (Ethics No. 2019-134). RESULTS: Wide-angle fundus photography and autofluorescence showed that the right eye was consistent with PPRCA and the left eye with RP. OCT showed that the outer layer of the fovea was intact in the right eye, while disorganized outer segment was found in the fovea of the left eye, and outer segment atrophies outside the fovea were found in both eyes. The amplitudes of ff-ERG decreased significantly in both eyes, and the amplitudes in right eye were slightly higher than those of the left eye. Visual field showed a paracentral arcuate scotoma in the right eye and severe centripetal contraction in the left eye. FFA showed hyperfluorescence in the retinal vein distribution area caused by atrophy of retinal pigment epithelium of the right eye and hypofluorescence related to bone spicule pigmentation, in addition with mottled hypofluorescence of choroid in the left eye. ICGA showed mild paravenous retinochroidal atrophy of the right eye and diffuse choroid capillaries atrophy in the middle and peripheral area of the left eye. WES revealed that the proband had a heterozygous c.2234C>T (p.Thr745Met) variant of the CRB1 gene. Sanger sequencing confirmed that the proband and family members except the father of the proband carried the same CRB1 gene variant. Based on the criteria and guidelines for the classification of genetic variation and related consensus from the American College of Medical Genetics and Genomics (ACMG), this variant was classified as pathogenic (PM3_VeryStrong+PM1+PM2_Supporting +PP3). CONCLUSION The heterozygous c.2234C>T (p.Thr745Met) variant of the CRB1 gene may underlay the unilateral PPRCA with contralateral eye RP in this proband. Above findings have enriched the mutational spectrum of the CRB1 gene.
Humans ; Electroretinography ; Exome Sequencing ; Eye Proteins/genetics* ; Membrane Proteins/genetics* ; Mutation ; Nerve Tissue Proteins/genetics* ; Pedigree ; Phenotype ; Retinitis Pigmentosa/genetics* ; Tomography, Optical Coherence ; Retinal Degeneration ; Eye Diseases, Hereditary

OBJECTIVE: To explore the clinical phenotype and genetic characteristics in two children with Knobloch syndrome (KNO) due to variants of COL18A1 gene. METHODS: Two children presented at the Genetic Eye Disease Clinic of the Eye Hospital of Wenzhou Medical University in October 2023 for ocular lesions were selected as the study subjects. Relevant clinical data and peripheral venous blood samples were collected from the children and their parents. Following genomic DNA extraction, whole-exome sequencing (WES) was carried out. Candidate variants were verified by Sanger sequencing of the family members. This study was approved by the Medical Ethics Committee of the hospital (Ethics No.: 2021-212-K-185). RESULTS: Both children exhibited characteristic ocular features of KNO including nystagmus, high myopia, and leopard spot fundus. Additionally, child 1 also presented with congenital occipital bone dysplasia and occipital encephalocele, while child 2 was diagnosed with vitreoretinochoroidopathy and bilateral high myopia. WES has identified compound heterozygous variants of the COL18A1 gene in both children, including a c.3013+3A>C splice-site variant and a c.2743C>T (p.Arg915Ter) nonsense variant in child 1, and a novel c.1702-1G>A splice-site variant and a c.3836C>T (p.Ser1279Leu) missense variant in child 2. A comprehensive literature review has identified 63 domestic and international articles involving 167 patients with KNO whom can be classified into three subtypes, with KNO type I being the most common and caused by pathogenic variants in the COL18A1 gene. Both probands in this study were children with KNO type I. Analysis of the genotype-phenotype correlations and population distribution characteristics revealed that the KNO patients exhibited significant clinical and genetic heterogeneity, along with a broad geographic distribution, with a relatively greater number of cases reported in Brazil and China. and a broad geographic distribution, with the highest numbers reported in Brazil and China. While no significant difference in genotype distribution was observed between Chinese and non-Chinese patients, phenotypic disparities were noted, with the non-Chinese cohort showing significantly higher rates of retinal detachment and developmental delay (P < 0.05), whereas Chinese patients exhibited a greater proportion of macular hypoplasia (P < 0.05). CONCLUSION The main clinical manifestations of KNO include high myopia, vitreoretinal dystrophy, and occipital encephalocele. The novel c.1702-1G>A splice-site variant identified in the COL18A1 gene has expanded the mutational spectrum of KNO type I and provided valuable insights for genetic diagnosis, counseling, and clinical management of the disease.
Humans ; Retinal Detachment/congenital* ; Male ; Female ; Child ; Encephalocele/genetics* ; Exome Sequencing ; Collagen Type XVIII/genetics* ; Phenotype ; Retinal Degeneration/genetics* ; Mutation ; Child, Preschool

Objective To study the application of autologous natural killer(NK)cells in the treatment of advanced renal cell carcinoma and the changes of immune function and tumor markers in patients.Methods 61 patients with advanced renal cell carcinoma admitted to Gansu Wuwei Tumour Hospital from March 2023 to April 2024 were divided into targeting group(31 cases,given supportive treatment combined with sunitinib malate capsules)and cell group(30 cases,autologous NK cells combined with targeting group)according to the patient's willingness to treat combined with propensity score matching.6 weeks was a treatment cycle,and all patients were treated for 4 cycles.The clinical efficacy of the two groups after 4 cycles of treatment and the occurrence of adverse reactions during treatment were statistically analyzed.The levels of serum cytokines,tumor markers,lymphocyte function and immune function were compared between the two groups before and after 4 cycles of treatment.Results After 4 cycles of treatment,the objective remission rate and disease control rate in the cell group were higher than those in the targeting group(P<0.05).After 4 cycles of treatment,the levels of serum hypoxia-inducible factor-1α(HIF-1α),vascular endothelial growth factor(VEGF),platelet-derived growth factor(PDGF),carcinoembryonic antigen,carbohydrate antigen 125,carbohydrate antigen 199 and alpha-fetoprotein in the two groups were lower than those before treatment,and those in the cell group were lower than those in the targeting group(P<0.012 5).After 4 cycles of treatment,the levels of serum interleukin-2,interleukin-6,tumor necrosis factor-β,interferon-γ,peripheral blood CD3+,CD4+,NK cells and CD4+/CD8+in the two groups were higher than those before treatment.The levels of serum interleukin-2,tumor necrosis factor-β,interferon-γ,peripheral blood CD3+and NK cells in the cell group were higher than those in the targeting group(P<0.012 5).The level of CD8+in peripheral blood was lower than that before treatment(P<0.012 5),but there was no significant difference between the two groups(P>0.012 5).During the treatment,there was no significant difference in the incidence of diarrhea,nausea and vomiting,alopecia,liver function damage,decreased platelet level and decreased neutrophil level between the cell group and the targeting group(P>0.05).Conclusion The treatment of advanced renal cell carcinoma with autologous NK cells could improve the level of serum cytokines,reduce the level of tumor markers,regulate the function of lymphocytes and immune function,and had a good therapeutic effect.At the same time,it would not increase the incidence of adverse reactions,and the safety was good.

Objective To investigate the ameliorative effects of the liver X receptor(LXR)agonist T0901317 on reti-nal structure and function damage caused by high glucose in mice.Methods A total of 30 12-week-old male C57BL/6J mice were randomly divided into the control,STZ-DMSO,and STZ-T0901317 groups,with 10 mice in each group.After a 12-hour fast,the mice were intraperitoneally injected with streptozotocin(STZ)at a dosage of 50 mg·kg-1 for 5 days.The mice in the control group were intraperitoneally injected with an equal volume of citrate buffer.The random blood glucose level was measured 72 hours after injection,and mice with the random blood glucose level exceeding 16.7 mmol·L-1 and polyuria were considered successful model candidates.These model mice were included in the STZ-DMSO and STZ-T0901317 groups,respectively,and the mice in the two groups were intravitreally injected with 1 μL of DMSO and 1 μL of T0901317(30 μmol·L-1),respectively.The thickness of the retinal nerve fiber layer(RNFL)was measured at 100 μm,200 μm,300 μm,400 μm,and 500 μm from the optic disc 6 weeks after STZ modeling.The cell density of the ganglion cell layer(GCL)(cells·mm-1)was quantified using ImageJ.Electroretinogram(ERG)analyses were employed to record the amplitude and latency of dark-adapted 0.01 ERG b-wave,dark-adapted 3.0 ERG a-and b-waves,dark-adapted 3.0 oscilla-tory potentials(OPs),and light-adapted 3.0 ERG a-and b-waves.The quantitative reverse transcription polymerase chain reaction(qRT-PCR)was used to detect the expression changes of inflammatory factors NLRP3,Caspase1,interleukin-1β(IL-1β),interleukin-6(IL-6),and tumor necrosis factor-α(TNF-α)in the retinal tissue of mice from each group.Results The optical coherence tomography(OCT)findings indicated that the RNFL thickness measured at 100 μm,200 μm,300μm,400 μm,and 500 μm from the optic disc in the STZ-DMSO group was significantly lower than that observed in the con-trol group,and the differences were statistically significant(all P＜0.05).In the STZ-T0901317 group,the RNFL thickness at all measurement points was greater than that in the STZ-DMSO group,and the differences were statistically significant(all P＜0.05).However,there were no statistically significant differences in the RNFL thickness at all measurement points between the STZ-T0901317 group and the control group(all P＞0.05).The hematoxylin and eosin(HE)staining results re-vealed that the GCL cell count in the control,STZ-DMSO,and STZ-T0901317 groups was(90.53±9.13)cells·mm-1,(82.06±0.98)cells·mm-1,and(94.94±10.61)cells·mm-1,respectively,with no significant differences among these groups(P＞0.05).The ERG results demonstrated that the amplitude of dark-adapted 0.01 ERG b-wave,dark-adapted 3.0 ERG a-and b-waves,the second wave of dark-adapted 3.0 oscillatory potentials(OPs2),and light-adapted ERG a-and b-waves was lower in the STZ-DMSO and STZ-T0901317 groups compared with the control group,and the differences were statistically significant(all P＜0.05).However,in the STZ-T0901317 group,the amplitude of dark-adapted 3.0 ERG a-and b-waves and OPs2 was higher than that in the STZ-DMSO group,and the differences were statistically significant(all P＜0.05).qRT-PCR analyses revealed that in the STZ-DMSO group,the relative mRNA expression level of NLRP3,Caspase-1,IL-1β,and IL-6 was elevated compared with the control group,and the differences were statistically significant(all P＜0.05).The relative mRNA expression level of NLRP3 and IL-6 in the STZ-T0901317 group was higher compared with the control group,and the differences were statistically significant(both P＜0.05).The relative mRNA expression level of NLRP3,Caspase-1,and IL-1β in the STZ-T0901317 group decreased compared with the STZ-DMSO group,and the differences were statistically significant(all P＜0.05).Conclusion The LXR agonist T0901317 may ameliorate early RNFL thickness alterations and im-prove visual function in DR mice by down-regulating the expression of NLRP3,Caspase-1,and IL-1β in mouse retinal tissues.

Objective To analyze the characteristics of death cases in a certain tertiary hospital,providing references for hospital management decisions and the evaluation of the effectiveness of medical services.Methods The information of death ca-ses of inpatients in the hospital from 2019 to 2023 was extracted through the inpatient medical records information system,and the medical records of inpatients were retrospectively analyzed by clinical epidemiological methods.Results The cumulative number of inpatients admitted to the hospital from 2019-2023 was 459 821,with 3 250 deaths and a case-fatality rate of 0.71％.There was a decreasing trend in the case fatality rate from 0.71％in 2019 to 0.63％in 2023,but the difference was not statistically significant(APC=-3.17,P＞0.05).The case fatality rate was 0.97％for males and 0.45％for females,with males having a higher case fatality rate than females(x2=435.206,P＜0.05).The mean age of the deceased cases was(69.59±17.70)years,and the case fatality rate increased gradually with age,and the highest case fatality rate was 2.98％in the age group of ≥80 years.The difference in case fatality rate between patients of different age groups was statistically significant(x2=3 502.991,P＜0.05).The top 5 causes of death of hospitalized patients were tumor,circulatory diseases,respiratory diseases,some infec-tious and parasitic diseases and injuries,poisoning and external causes,accounting for 87.33％of the total deaths.The main causes of death in neoplasms were bronchial and pulmonary malignancies,hepatic and intrahepatic biliary malignancies,and gas-tric malignancies;in the circulatory diseases,cerebral infarction,acute myocardial infarction,and intracerebral haemorrhage;and in the respiratory diseases,pathogens unspecified pneumonias,bacterial pneumonias,and other chronic obstructive pulmona-ry diseases.Conclusion The case fatality rate of male and ≥80 years old patients in this hospital is higher.Tumor,circulatory diseases and respiratory diseases are the main causes of death.The hospital should allocate medical resources in a scientific and reasonable way according to the age,gender and disease characteristics of the patients;strengthen the prevention and treatment of tumors,circulatory diseases and respiratory diseases according to the cause of death and their distribution,improve the diagnosis and treatment level,and reduce the inpatient morbidity and fatality rate.

Objective To explore the effect of visual balance training combined with digital treadmill intervention on walking func-tion in patients with ischemic stroke.Methods From July,2023 to December,2024,90 patients with ischemic stroke in Gansu Provincial Hospital were ran-domly divided into control group(n=30),treadmill group(n=30)and combined group(n=30).All groups re-ceived routine rehabilitation treatment,while the treadmill group added digital treadmill training,and the com-bined group added visual feedback balance training and digital treadmill training,for four weeks.All groups were assessed with Berg Balance Scale(BBS),Pro-Kin visual feedback balance training system,Tinetti Performance-Oriented Mobility Assessment(POMA),digital treadmill system,Timed Up and Go Test(TUGT),Fugl-Meyer Assessment-Lower Extremities(FMA-LE),Functional Ambulation Category(FAC)and modified Barthel Index(MBI)before and after intervention.Results The effects of intra-group(F>147.291,P<0.001),inter-group(F>4.919,P<0.05)and interaction(F>18.386,P<0.001)were all significant for the indicators including BBS score,length trajectory and elliptical area of eyes open or closed,POMA score,step length,hip and knee range of motion on the healthy and affected side,TUGT time,FMA-LE score,and MBI score.Post-hoc tests showed that after treatment,all the above indicators improved in each group(P<0.01),and they were the best in the combined group,followed by the treadmill group(P<0.05).After treatment,the FAC grades improved in all the groups(|Z|>1.971,P<0.05),and it was better in the combined group than in the control group(P<0.01).Conclusion Visual feedback balance training combined with digital treadmill intervention can improve balance function,walking ability and activities of daily living in patients with ischemic stroke,which is more effective than tread-mill training alone.

Objective To analyze the characteristics of death cases in a certain tertiary hospital,providing references for hospital management decisions and the evaluation of the effectiveness of medical services.Methods The information of death ca-ses of inpatients in the hospital from 2019 to 2023 was extracted through the inpatient medical records information system,and the medical records of inpatients were retrospectively analyzed by clinical epidemiological methods.Results The cumulative number of inpatients admitted to the hospital from 2019-2023 was 459 821,with 3 250 deaths and a case-fatality rate of 0.71％.There was a decreasing trend in the case fatality rate from 0.71％in 2019 to 0.63％in 2023,but the difference was not statistically significant(APC=-3.17,P＞0.05).The case fatality rate was 0.97％for males and 0.45％for females,with males having a higher case fatality rate than females(x2=435.206,P＜0.05).The mean age of the deceased cases was(69.59±17.70)years,and the case fatality rate increased gradually with age,and the highest case fatality rate was 2.98％in the age group of ≥80 years.The difference in case fatality rate between patients of different age groups was statistically significant(x2=3 502.991,P＜0.05).The top 5 causes of death of hospitalized patients were tumor,circulatory diseases,respiratory diseases,some infec-tious and parasitic diseases and injuries,poisoning and external causes,accounting for 87.33％of the total deaths.The main causes of death in neoplasms were bronchial and pulmonary malignancies,hepatic and intrahepatic biliary malignancies,and gas-tric malignancies;in the circulatory diseases,cerebral infarction,acute myocardial infarction,and intracerebral haemorrhage;and in the respiratory diseases,pathogens unspecified pneumonias,bacterial pneumonias,and other chronic obstructive pulmona-ry diseases.Conclusion The case fatality rate of male and ≥80 years old patients in this hospital is higher.Tumor,circulatory diseases and respiratory diseases are the main causes of death.The hospital should allocate medical resources in a scientific and reasonable way according to the age,gender and disease characteristics of the patients;strengthen the prevention and treatment of tumors,circulatory diseases and respiratory diseases according to the cause of death and their distribution,improve the diagnosis and treatment level,and reduce the inpatient morbidity and fatality rate.