Objective To investigate the impact of mitophagy,mediated by the long non-coding RNA SNHG16(LncRNA SNHG16)on diabetes-associated cognitive impairment(DCI).Methods 29 male C57BL/J mice were randomly divided into normal control(NC)group,DCI group and DCI+mitochondrial autophagy inhibitor(DCI+Mdivi-1)group.Morris water maze and new object recognition test were used to detect the cognitive function of mice,qRT-CPR was used to detect the expression of LncRNA SNHG16 and mitochondrial autophagy marker mRNA.Western blot were used to detect the expression of related protein.The mouse hippocampal neurons HT22 were divided into control(Con)group,high glucose(HG)group,HG+SNHG16 silencing(HG+sh-SNHG16)group and HG+no-load control(HG+sh-NC)group.CCK8 method and lactate dehydrogenase(LDH)method were used to detect neuronal damage.JC-1 method was used to detect mitochondrial membrane potential.Results Compared with NC group,the expression of LncRNA SNHG16 and the expression of autophagy-related gene 5,PTEN-induced putative kinase 1(PINK1),Parkin and microtubule associated protein light chain 3(LC3)Ⅱ/Ⅰ increased(P<0.05),while the expression of mitochondrial autophagy-related proteins P62 and mitochondrial outer membrane transposase 20(TOMM20)decreased in T2DM group.Compared with DCI group,the cognitive dysfunction of mice improved,and the expression level of LncRNA SNHG16 decreased in the DCI+Mdivi-1 group(P<0.05).The expressions of LncRNA SNHG16,LC3 Ⅱ/Ⅰ,PINK1 and Parkin were higher in HG group than in Con group(P<0.05),while the cell survival rate and TOMM20 protein expression were lower in HG group than in Con group(P<0.05).Silence of LncRNA SNHG16 can restore the activity of HT22 cells and mitochondrial membrane potential,and reduce the level of mitochondrial autophagy under HG condition.Conclusions The expression level of LncRNA SNHG16 was up-regulated in the hippocampus brain region of mice with diabetic cognitive dysfunction,and mitophagy was overactivated.Silencing of LncRNA SNHG16 inhibits mitophagy in hippocampal neurons and alleviates HG induced hippocampal neuronal damage.

Objective To investigate the impact of mitophagy,mediated by the long non-coding RNA SNHG16(LncRNA SNHG16)on diabetes-associated cognitive impairment(DCI).Methods 29 male C57BL/J mice were randomly divided into normal control(NC)group,DCI group and DCI+mitochondrial autophagy inhibitor(DCI+Mdivi-1)group.Morris water maze and new object recognition test were used to detect the cognitive function of mice,qRT-CPR was used to detect the expression of LncRNA SNHG16 and mitochondrial autophagy marker mRNA.Western blot were used to detect the expression of related protein.The mouse hippocampal neurons HT22 were divided into control(Con)group,high glucose(HG)group,HG+SNHG16 silencing(HG+sh-SNHG16)group and HG+no-load control(HG+sh-NC)group.CCK8 method and lactate dehydrogenase(LDH)method were used to detect neuronal damage.JC-1 method was used to detect mitochondrial membrane potential.Results Compared with NC group,the expression of LncRNA SNHG16 and the expression of autophagy-related gene 5,PTEN-induced putative kinase 1(PINK1),Parkin and microtubule associated protein light chain 3(LC3)Ⅱ/Ⅰ increased(P<0.05),while the expression of mitochondrial autophagy-related proteins P62 and mitochondrial outer membrane transposase 20(TOMM20)decreased in T2DM group.Compared with DCI group,the cognitive dysfunction of mice improved,and the expression level of LncRNA SNHG16 decreased in the DCI+Mdivi-1 group(P<0.05).The expressions of LncRNA SNHG16,LC3 Ⅱ/Ⅰ,PINK1 and Parkin were higher in HG group than in Con group(P<0.05),while the cell survival rate and TOMM20 protein expression were lower in HG group than in Con group(P<0.05).Silence of LncRNA SNHG16 can restore the activity of HT22 cells and mitochondrial membrane potential,and reduce the level of mitochondrial autophagy under HG condition.Conclusions The expression level of LncRNA SNHG16 was up-regulated in the hippocampus brain region of mice with diabetic cognitive dysfunction,and mitophagy was overactivated.Silencing of LncRNA SNHG16 inhibits mitophagy in hippocampal neurons and alleviates HG induced hippocampal neuronal damage.

ObjectiveTo investigate the effect of kinesin family member 15 （KIF15） on the proliferation of hepatocellular carcinoma （HCC） cells and its mechanism of action. MethodsTCGA and GEPIA datasets were analyzed to determine the expression of KIF15 in HCC and its effect on tumor stage and survival. Quantitative real-time PCR and Western blot were used to measure the expression level of KIF15 in human-derived HCC cell lines （HepG2， Hep3B， MHCC-97H， and LM3） and human normal liver cell line L02 cultured in vitro， and Hep3B and HepG2 were selected for subsequent studies. CCK-8 assay， plate colony formation assay， and EdU staining were performed for Hep3B cells transfected with shRNA-NC or shRNA-KIF15 and HepG2 cells transfected with LV-vector or LV-KIF15 to evaluate the viability and proliferative capacity of these cells. GSEA was used to analyze the potential signaling pathways associated with KIF15 in HCC， and Western blot was used for detection. The independent-samples t test was used for comparison of continuous data between two groups； a one-way analysis of variance was used for comparison between multiple groups， and the least significant difference t-test was used for further comparison between two groups. ResultsThe analysis of TCGA and GEPIA datasets showed that in HCC patients， the expression of KIF15 in HCC tissue was significantly higher than that in normal tissue， and the HCC patients with high KIF15 expression tended to have a poorer prognosis. Compared with sh-NC-Hep3B， sh3-Hep3B showed significant reductions in the mRNA and protein levels of KIF15 （P<0.05）， cell viability， clone formation number， and EdU positive rate （all P<0.05）. Compared with vector-HepG2， LV-KIF15-HepG2 showed significant increases in the mRNA and protein levels of KIF15 （P<0.05）， cell viability， clone formation number， and EdU positive rate （all P<0.05）. Subcutaneous tumor assay showed that compared with sh-NC-Hep3B， sh3-Hep3B showed reductions in tumor volume and tumor weight， as well as a significant reduction in the immunohistochemical score of Ki67 and a significant increase in the immunohistochemical score of TUNEL （P<0.05）. GSEA analysis showed that the PI3K/AKT/mTOR pathway was positively correlated with KIF15 in HCC （NES=1.59， P<0.001）. Western blot showed that LY294002 could inhibit the PI3K/AKT/mTOR pathway upregulated in LV-KIF15-HepG2， and compared with LV-KIF15-HepG2， LY294002+LV-KIF15-HepG2 showed significant reductions in cell viability， clone formation number， and EdU positive rate （all P<0.05）. ConclusionKIF15 enhances the viability and proliferative capacity of HCC cells by upregulating the PI3K/AKT/mTOR signaling pathway.

AIM:The objective of this study was to investigate the ameliorative effect of Erjing pills(PL)on the functional impairment of the diabetic retinal neurovascular unit(NVU)in rats.METHODS:A diabetic retinopathy(DR)model was established using BN rats,which were then treated with freeze-dried Erjing pills powder.After 12 weeks,various assessments were conducted:retinal thickness was measured using OCT,fundus lesions were evaluated with FFA,and retinal cell structure,arrangement,and layer thickness were analyzed through HE staining.Additionally,transmission electron microscopy was employed to examine ultrastructural changes in retinal ganglion cells(RGCs),and retinal leakage was assessed using EB in vivo perfusion.The expression of ZO-1,a tight junction protein in the retina,was detected via immunohistochemistry,while Western blot analysis quantified the levels of ZO-1,occludin,and claudin-5 proteins in rat retinal tissue.Serum vascular endothelial growth factor(VEGF)levels were measured using an ELISA kit.RESULTS:Results showed that OCT,FFA,HE staining,and transmission electron microscopy indicated a significant reduction in retinal thickness in the MOD group compared to the CON group(P＜0.05).In contrast,the retinal thickness increased in the CDC,PLL,and PLH groups compared to the MOD group,with significant improvement observed in the PLH group(P＜0.05).Additionally,the number of RGCs increased,and sodium fluorescein leakage from fundus mi-crovessels decreased in the CON group relative to the administered groups.Transmission electron microscopy revealed ir-regular deformation of RGC nuclei,mitochondrial reduction and swelling,and organelle loss in the MOD group compared to the CON group.However,the CDC,PLL,and PLH groups demonstrated varying degrees of improvement in cellular layers.EB leakage assay results indicated a significant increase in retinal EB leakage in the MOD group compared to the CON group(P＜0.01).EB leakage was significantly reduced in the CDC group compared to the MOD group(P＜0.01),with some reduction also observed in the PLL and PLH groups(P＜0.05,P＜0.01).Immunohistochemistry and Western blot analyses revealed a significant decrease in the expression of retinal tight junction proteins ZO-1,occludin,and clau-din-5 in the MOD group compared to the CON group(P＜0.05).Protein expression was notably higher in the CDC group compared to the MOD group(P＜0.01),with increases also observed in the PLL and PLH groups(P＜0.05,P＜0.01).ELISA results showed that VEGF levels were significantly higher in the MOD group compared to the CON group(P＜0.01).The CDC,PLL,and PLH groups exhibited varying degrees of reduction in VEGF levels compared to the MOD group(P＜0.05,P＜0.01).CONCLUSION:These findings suggest that PL effectively ameliorates retinal NVU damage in DR rats.

AIM:The objective of this study was to investigate the ameliorative effect of Erjing pills(PL)on the functional impairment of the diabetic retinal neurovascular unit(NVU)in rats.METHODS:A diabetic retinopathy(DR)model was established using BN rats,which were then treated with freeze-dried Erjing pills powder.After 12 weeks,various assessments were conducted:retinal thickness was measured using OCT,fundus lesions were evaluated with FFA,and retinal cell structure,arrangement,and layer thickness were analyzed through HE staining.Additionally,transmission electron microscopy was employed to examine ultrastructural changes in retinal ganglion cells(RGCs),and retinal leakage was assessed using EB in vivo perfusion.The expression of ZO-1,a tight junction protein in the retina,was detected via immunohistochemistry,while Western blot analysis quantified the levels of ZO-1,occludin,and claudin-5 proteins in rat retinal tissue.Serum vascular endothelial growth factor(VEGF)levels were measured using an ELISA kit.RESULTS:Results showed that OCT,FFA,HE staining,and transmission electron microscopy indicated a significant reduction in retinal thickness in the MOD group compared to the CON group(P＜0.05).In contrast,the retinal thickness increased in the CDC,PLL,and PLH groups compared to the MOD group,with significant improvement observed in the PLH group(P＜0.05).Additionally,the number of RGCs increased,and sodium fluorescein leakage from fundus mi-crovessels decreased in the CON group relative to the administered groups.Transmission electron microscopy revealed ir-regular deformation of RGC nuclei,mitochondrial reduction and swelling,and organelle loss in the MOD group compared to the CON group.However,the CDC,PLL,and PLH groups demonstrated varying degrees of improvement in cellular layers.EB leakage assay results indicated a significant increase in retinal EB leakage in the MOD group compared to the CON group(P＜0.01).EB leakage was significantly reduced in the CDC group compared to the MOD group(P＜0.01),with some reduction also observed in the PLL and PLH groups(P＜0.05,P＜0.01).Immunohistochemistry and Western blot analyses revealed a significant decrease in the expression of retinal tight junction proteins ZO-1,occludin,and clau-din-5 in the MOD group compared to the CON group(P＜0.05).Protein expression was notably higher in the CDC group compared to the MOD group(P＜0.01),with increases also observed in the PLL and PLH groups(P＜0.05,P＜0.01).ELISA results showed that VEGF levels were significantly higher in the MOD group compared to the CON group(P＜0.01).The CDC,PLL,and PLH groups exhibited varying degrees of reduction in VEGF levels compared to the MOD group(P＜0.05,P＜0.01).CONCLUSION:These findings suggest that PL effectively ameliorates retinal NVU damage in DR rats.

Since there is a lack of obvious clinical symptoms in the early stage of hepatocellular carcinoma (HCC), most patients have progressed to the advanced stage at the time of confirmed diagnosis. There are limited treatment options for HCC patients who miss the opportunity for surgery, so it is of great importance to find new therapeutic targets. Tumor-associated macrophages (TAMs) are a group of macrophages existing in the tumor immune microenvironment and affect the malignant behaviors of HCC cells and the state of immune escape within the tumor. This article introduces the origin and classification of TAM, summarizes the role and mechanism of TAMs in vascular proliferation, invasion and metastasis, formation and maintenance of stemness, and anti-tumor immunity in HCC, and briefly describes the current research advances in therapeutic targets for TAM, and it is pointed out that targeting TAM may be a promising direction for clinical treatment.

OBJECTIVE: To explore the genetic basis for a child featuring short stature and postaxial polydactyly. METHODS: A child who presented at Ningbo Women & Children's Hospital in May 2021 due to the"discovery of growth retardation for more than two years" was selected as the subject. Peripheral blood samples of the child and his parents were collected for the extraction of genomic DNA. Whole exome sequencing was carried out for the child, and candidate variant was verified by Sanger sequencing of his family members. RESULTS: The child was found to harbor a heterozygous c.3670C>T (p.Q1224) variant of the GLI2 gene, which may lead to premature termination of protein translation. The variant was not detected in either parent. CONCLUSION The child was diagnosed with Culler-Jones syndrome. The c.3670C>T (p.Q1224*) variant of the GLI2 gene probably underlay the disease in this child.
Child ; Female ; Humans ; Fingers ; Mutation ; Nuclear Proteins/genetics* ; Polydactyly/genetics* ; Toes ; Zinc Finger Protein Gli2/genetics*

Abstract On April 25, 2021, a case with hookworm infections was identified during the surveillance of soil-borne nematodiasis by Tiantai Center for Disease Control and Prevention, Taizhou City, Zhejiang Province. The patients was admitted to a county people's hospital due to dizziness and limb weakness on July 24, 2020. Laboratory tests showed 2.5×1012/L red blood cell counts, 45 g/L hemoglobin, 3.4% eosinophil percentage, 4.7 μmol/L serum iron and <1 μg/L ferritin, and severe iron-deficiency anemia was initially diagnosed. Following treatment with blood transfusion and ion supplement, subsequent three routine blood tests indicated elevated eosinophil percentages. On April 25, 2021, hookworm eggs were detected in stool samples using Kate-Katz technique, and Necator americanus was identified with the test-tube filter-paper culture method. Severe iron-deficiency anemia caused by hookworm infections was diagnosed based on field epidemiological surveys and laboratory tests. The health education pertaining to parasitic disease control knowledge among residents and the diagnosis and treatment of parasitic diseases in medical institutions are recommended to be improved in rural areas to avoid misdiagnosis and missing diagnosis.

Objective:To explore the effect of aquaporin 4 (AQP4) regulated by miR-320a on a cell model of Alzheimer′s disease.Methods:A rat adrenal pheochromocytoma cell line (PC12) was induced into neurons using nerve growth factor (NGF). The morphology of the PC12 cells and the neurons was observed, and ubiquitin carboxy terminal hydrolase L1 (Uch-L1) and neurofilament protein (NFP) were detected. Levels of microtubule-associated protein (MAP2) and AQP4 target genes were related to the mRNA expression of NFP to determine the neuron-inducing effect. The neurons were then randomly divided into a control group (given no treatment), an miR-320a mimic transfection group (cultured by adding 50nmol/L miR-320a as a mimic agent), an miR-320a inhibitor group (cultured by adding 50nmol/L miR-320a as an inhibitor), an Aβ treatment group (cultured by adding Aβ), an Aβ+ miR-320a mimic group (cultured by adding both 50nmol/L miR-320a and Aβ), and an Aβ+ miR-320a inhibitor group (also cultured by adding Aβ, but with 50nmol/L miR-320a as an inhibitor). Cell activity was measured by the CCK8 method. Reverse-transcription polymerase chain reactions were used to detect the relative expression of the target gene miR-320a, AQP4, B-cell bcl2-associated X (Bax), and B-cell bcl-2 (Bcl-2) mRNA. Western blotting was employed to detect the relative expression of AQP4, Bax and Bcl-2 proteins.Results:After PC12 was induced by 50μg/L NGF, the expression of Uch-L1 genes in the induced neuron was significantly down-regulated compared with the PC12. The expressions of NFP, MAP2 and AQP4 genes were significantly up-regulated, and the relative expressions of MAP2 and AQP4 proteins increased significantly. Compared with the control group, the apoptosis and cell activity of neurons in the treatment group increased, the mRNA and protein expressions of miR-320a, AQP4, bcl-2, AQP4 and Bcl-2 decreased significantly, and the mRNA and protein expressions of Bax increased significantly. Compared with the Aβ-treated group, the cell activity of the Aβ+ Mir-320a mimic group increased significantly, the mRNA and protein expressions of miR-320a, AQP4 and Bcl-2 increased significantly, and the mRNA and protein expressions of Bax decreased significantly. Compared with the Aβ+ miR-320a mimic group, the cell activity of the Aβ+ miR-320a inhibitor group decreased significantly, the mRNA and protein expressions of miR-320a, AQP4 and Bcl-2 decreased significantly, and the mRNA and protein expressions of Bax increased significantly.Conclusion:miR-320a can up-regulate AQP4 expression in a cell model of Alzheimer′s disease, reduce apoptosis and increase the cell survival rate.

Objective: To explore the value of chest CT features and clinical indexes in the differential diagnosis between suspected COVID-19 with two or more negative nucleic acid tests and confirmed COVID-19. Methods: The clinical data and chest CT images of 105 cases withconfirmedCOVID-19 (55 males and 50 females, aged from 2 month to 88 years) and 97 cases with suspected COVID-19(59 males and 38 females, aged from 1 month to 93 years) were analyzed retrospectively in Shiyan Taihe Hospital from January 21 to February 10, 2020. χ²test and two independent sample t test were used to analyze the clinical data and CT signs of the two cases, with P<0.05 for the difference statistically significant. Results: Compared with the suspected patients, the average age of diagnosis of covid-19 was higher (t = 2.460, P = 0.01). The main pathological changes were pure ground glass (68 cases) and mixed ground glass density (53 cases) (χ² = 50.016, P< 0.01). Interstitial thickening (83 cases) (χ² = 55.395, P< 0.01), vascular thickening (73 cases) (χ² = 57.527, P< 0.01), air bronchoscopic sign or bronchiectasis Zhang (67 cases) (χ² = 17.899, P< 0.01), cord focus (54 cases) (χ² = 5.500, P = 0.02), easily distributed under the pleura and the long axis of the lesion was parallel to the pleura (89 cases) (χ² = 23.597, P< 0.01), most of them had no pleural effusion (χ² = 7.017, P< 0.01); both lesions were mainly distributed in patches (89 cases were confirmed, 87 suspected) (χ² = 19.573, P< 0.01). In addition, the lesions of patients with confirmed covid-19 showed progress in short term (72 / 87, 82.76%), and those with suspected covid-19 showed remission in short term (63 / 89, 70.78%). The difference was statistically significant (χ² = 51.114, P< 0.01). There was no significant difference in gender and distribution of pulmonary lobes (χ² = 1.462, P= 0.23; χ² = 7.381, P= 0.19). The number of white blood cells (χ² = 17.891, P< 0.01) and the percentage of lymphocytes (χ² = 11.151,P< 0.01) of covid-19 were mostly normal or decreased, creatine kinase (χ² = 9.589, P< 0.01) were mostly normal, and erythrocyte sedimentation rate was mostly normal or increased (χ² = 4.240, P= 0.04). Conclusions The imaging features and biochemical indexes of diagnosed COVID-19 are different from those of suspected COVID-19. The comparative analysis of imaging features, clinical indexes and reexamination are helpful for the differential diagnosis of COVID-19 and suspected COVID-19.