Objective To study the expression of Ca-A/K channel-related molecules glutamate receptor 2 and 1(GluR2/1) in hippocampus tissues of neonatal rats with hypoxic-ischemic brain damage (HIBD). Methods A total of 60 7-day-old Sprague-Dawley rats were randomly divided into sham operation group and HIBD group. Hippocampal tissues were obtained at 0 h, 1 h, 6 h, 24 h, 48 h and 72 h after HIBD. The expression of GluR2, GluR1 and autophagy marker protein Beclin-1, LC3 were detected by Western blot assay. Results Edema and focal softening and necrosis were observed 6 h after HIBD in the brains of neonatal rats. Compared with Con group, at each time point, the expression levels of GluR2 were lower while the levels of GluR1, Beclin-1 and LC3 were higher significantly in HIBD group (P<0.05). The protein levels of LC3, Beclin-1, GluR1 and GluR2 in hippocampus tissues of HIBD group were significantly different among different time points after the estab-lishment of HIBD model (F=10.65~701.14, P<0.01). The protein level of GluR2 was decreased from 1 h to 24 h after HIBD and reached the lowest level at 24 h. The levels of GluR1, Beclin-1 and LC3 were increased at 6 h, plateaued at 24 h and remained there until 48 h. The levels of these proteins returned back to the initial level at 72 h. Conclusions Ca-A/K channel-related mol-ecules GluR2 and GluR1 play important roles in the autophagic cell death of hippocampus tissues in neonatal rats with hypoxic-ischemic brain damage.

Objective To study the effects of melatonin (MT) on mitochondrial autophagy in neonatal rats with hypoxic-ischemic brain damage (HIBD).Method Animal model of HIBD was established.Forty-five 7-day-old Sprague-Dawley (SD) rats were randomly assigned to sham operation group and HIBD group.Brain tissue were taken at 0,2,4,6,8,12,24 and 48 h after model preparation,and the expressions of mitochondrial autophagy-related protein Bnip3 and autophagy-related protein LC3-Ⅱ were detected.Seventy-two 7-day-old SD rats were randomly assigned to sham operation group,HIBD group and post-HIBD treatment group (3-MA,Mdivi-I,Rapa,MT,3-MA + MT,Mdivi-1 + MT,Rapa + MT).The sizes of cerebral infarction after different treatment were detected using triphenyltetrazolium chloride staining (TIC).Primary cortical cells of fetal SD rats (embryonic day:17 ～ 19 d) were cultured.JC-I staining was used to detect mitochondrial membrane potential and immunofluorescence method was used to observe mitochondrial autophagy.The Oxygen glucose deprivation/reperfusion/R (OGD) model was prepared.Autophagy inhibitor 3-MA,mitochondrial autophagy inhibitor Mdivi-1,autophagy activator Rapa,and MT were applied and Bnip3 and LC3-Ⅱ expressions and CCK8 (Cell Counting Kit CCK 8) for cell viability assay were examined.Result TTC staining results showed significant white infarcts in the tissue of HIBD group after hypoxia-ischemia,especially in the 3-MA and Mdivi-1 groups,and the infarcts were smaller in Rapa group and groups with MT treatment,the differences were statistical significant (P ＜ 0.05).Compared with the sham operation group,the expressions of Bnip3 and LC3-Ⅱ in the HIBD group were significantly increased (P ＜ 0.05).Compared with the normal group,the expressions of Bnip3 and LC3-Ⅱ in the OGD/R group were increased (P ＜0.05).The activities of 3-MA and Mdivi-1 cells decreased significantly,the mitochondrial membrane potential decreased,and mitochondrial autophagy were decreased (P ＜ 0.05).The cell activity,mitochondrial membrane potential,and mitochondrial autophagy of Rapa group were increased (P ＜ 0.05).The cell viability,Bnip3 and LC3-Ⅱ expressions were increased in groups with MT intervention (P ＜ 0.05).Conclusion MT may play an important protective role in the early stage of brain injury by enhancing mitochondrial autophagy of HIBD,which provide a theoretical basis for the study of specific related mechanisms.

OBJECTIVETo explore the correlation between 13q33-q34 microdeletion and clinical phenotype.

METHODSRoutine chromosomal banding was performed to analyze the karyotype, while array-based comparative genomic hybridization (aCGH array) and single nucleotide polymorphism array(SNP array) were employed to investigate the genome copy number variations.

RESULTSThe karyotype of patient 1 was 46, XY, 9qh+,13qs. Patient 2 showed 46, XX, der (13). Patient 3 showed 46, XX, r(13) (p11.2q32) [43]/45, XX, 13[4]/46, XX, r(13;13) [2]/47, XX, 2r(13;13) [1]. Patient 4 did not undergo chromosome karyotyping analysis. Array analysis showed that four patients have different microdeletions in 13q33-34 region and had common features of 13q33-q34deletion including intellectual disability, facial dysmorphism, microcephaly, hypotonia, low birth weight and genital abnormality.

CONCLUSIONThe severity of phenotypes showed no correlation with the size of deletion in 13q33-q34. The lower percentage of patients with congenital heart disease suggested a complex pathogenesis of such disease. EFNB2, LIG4 and SOX1 in 13q33-34 region are promising candidates for mental retardation. LIG4 was also a likely candidate for microcephaly.

Child, Preschool ; Chromosome Banding ; methods ; Chromosome Deletion ; Chromosomes, Human, Pair 13 ; genetics ; Female ; Genetic Testing ; methods ; Humans ; Infant ; Intellectual Disability ; genetics ; Male

This study aimed to evaluate the efficacy and safety of mycophenolate mofetil (MMF) or tacrolimus (TAC) compared with azathioprine (AZA) as maintenance therapy for active lupus nephritis (ALN). Patients with ALN who responded to 24 weeks of induction treatment were enrolled. Patients who received MMF or TAC as induction therapy continued MMF or TAC treatment during the maintenance period, whereas those who received intravenous cyclophosphamide were subjected to AZA treatment. The primary endpoint was the incidence of renal relapse. Secondary endpoints included extrarenal flares and composite endpoints (deaths, end-stage renal disease, or doubling of serum creatinine levels). A total of 123 ALN patients (47 in the MMF group, 37 in the TAC group, and 39 in the AZA group) were enrolled. The median follow-up time was 60 months. Ten MMF-treated patients, ten TAC-treated patients, and eight AZA-treated patients experienced renal relapses (P = 0.844). The cumulative renal relapse rates in the MMF group (P = 0.934) and TAC group (P = 0.673) were similar to the renal relapse rate in the AZA group. No significant difference in the incidence of severe adverse event was observed among the groups. Long-term maintenance therapies with MMF or TAC might have similarly low rates of renal relapse and similar safety profiles compared with AZA.
Humans ; Mycophenolic Acid/adverse effects* ; Azathioprine/adverse effects* ; Tacrolimus/therapeutic use* ; Lupus Nephritis/complications* ; Immunosuppressive Agents ; Treatment Outcome ; Recurrence

The development of broad-spectrum antivirals against human coronaviruses (HCoVs) is critical to combat the current coronavirus disease 2019 (COVID-19) pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and its variants, as well as future outbreaks of emerging CoVs. We have previously identified a polyethylene glycol-conjugated (PEGylated) lipopeptide, EK1C4, with potent pan-CoV fusion inhibitory activity. However, PEG linkers in peptide or protein drugs may reduce stability or induce anti-PEG antibodies in vivo. Therefore, we herein report the design and synthesis of a series of dePEGylated lipopeptide-based pan-CoV fusion inhibitors featuring the replacement of the PEG linker with amino acids in the heptad repeat 2 C-terminal fragment (HR2-CF) of HCoV-OC43. Among these lipopeptides, EKL1C showed the most potent inhibitory activity against infection by SARS-CoV-2 and its spike (S) mutants, as well as other HCoVs and some bat SARS-related coronaviruses (SARSr-CoVs) tested. The dePEGylated lipopeptide EKL1C exhibited significantly stronger resistance to proteolytic enzymes, better metabolic stability in mouse serum, higher thermostability than the PEGylated lipopeptide EK1C4, suggesting that EKL1C could be further developed as a candidate prophylactic and therapeutic for COVID-19 and other coronavirus diseases.