ObjectiveTo evaluate the screening performance of commonly used renal function indicators for chronic kidney disease (CKD) in subjects of different ages, so as to explore the appropriate screening regimen for Chinese population.MethodsA total of 2 131 adult subjects in Sichuan Orthopaedic Hospital or Mianyang Central Hospital from May 2016 to October 2017 were selected. They were divided into three groups according to age: group A (18–39 years, n=278), group B (40–64 years, n=1 167), and group C (≥65 years, n=686). Serum levels of creatinine (SCr), urea, and cystatin C [CysC; to calculate estimated glomerular filtration rate (eGFR) based on this index], as well as urine levels of albumin, total protein and creatinine were measured, and urine albumin to creatinine ratio (ACR) and urine protein to creatinine ratio (PCR) were calculated respectively. CKD was diagnosed according to the Kidney Disease: Improving Global Outcomes (KDIGO) Guideline (2012 Edition). The receiver-operating characteristic (ROC) curve analysis was finally performed to investigate the diagnostic performance of each indicator for CKD.ResultsThe prevalences of CKD in group A, B, and C were 10.8% (30/278), 16.4% (191/1 167), and 45.8% (314/686), respectively, and the difference among these groups was statistically significant (χ2=233.525, P<0.001). In addition, the levels of the six renal function indicators between CKD and non-CKD subjects also had statistically significant differences in different age groups (P<0 05="" otherwise="" roc="" curve="" analysis="" revealed="" that="" the="" diagnostic="" values="" of="" these="" indicators="" were:="" acr="" or="" pcr=""> eGFR or CysC > serum urea or SCr (AllP<0 05="" except="" that="" egfr="" cysc="" serum="" urea="" and="" scr="" in="" group="" a="" assessed="" the="" similar="" diagnostic="" performance="" among="" the="" three="" indicators="" recommended="" by="" kdigo="" guideline="" the="" diagnostic="" performances="" of="" acr="" and="" pcr="" in="" different="" age="" groups="" were:="" group="" b="" 0="" 915="" 0="" 914=""> group A (0.885, 0.890) > group C (0.841, 0.846), while the diagnostic performance of eGFR was group C (0.807) > group B (0.728) > group A (0.696). The best boundary values of ACR and PCR were lower while the value of eGFR was higher than the corresponding KDIGO recommended medical decision levels.ConclusionsACR is the first choice for screening CKD when the renal function index creatinine is normal. Moreover, eGFR can further improve the diagnostic value, especially in subjects over 65 years old.
Objective To find a new specific marker that can be used to early diagnose hepatic fibrosis by detecting the change of serum protein in patients with hepatic fibrosis. Methods This research adopted 50 SD rats (25 males and 25 females), and from which 6 rats were selected randomly (3 males and 3 females) as control group, last 44 rates were divided into four groups according to four pathological stages as hepatic fibrosis model group (experimental group). Distinct proteins in serum were detected by surface-enhanced laser desorption/ionization-time of flight-mass spectra (SELDI-TOF-MS). Radioimmunoassay was used to measure four parameters of hepatic fibrosis which were hyaluronidase (HA), precollagen Ⅲ (PCⅢ), laminin (LN) and collagen Ⅳ (Ⅳ-C). Results Distinct proteins in serum were detected in 8 cases of stage Ⅰ of hapatic fibrosis, 5 cases of stage Ⅱ, 5 cases of stage Ⅲ, 6 cases of stage Ⅳ, and 5 cases of control by SELDI-TOF-MS. Three protein peaks were found (M/Z: 4 203, 4 658, and 7 400). The peaks of M/Z 4 658 and 4 700 proteins were obviously increased in the stage Ⅰ of hepatic fibrosis （Plt;0.05）, while the changes of hepatic fibrosis four parameters appeared in stage Ⅳ of hepatic fibrosis. Conclusion This method shows great potential for early diagnosing of hepatic fibrosis and finding better biomarkers to hepatic fibrosis.
Objective To investigate the cellular viability and mitochondrial reactive oxygen species (ROS) production of the Müller cells under high glucose condition, and explore the protection role of the 5,6-dihydrocyclopenta-1, 2-dithiole-3-thione (CPDT) on Müller cells. Methods Müller cells from Sprague Dawley rats were divided into 5 groups randomly, including 25 mmol/L normal glucose group (group A) and 65 mmol/L high glucose group (group B). High glucose group with 45, 60, 70 μmol/L CPDT and cultured them 72 hour was set as group C, D and E. Water soluble tetrazolium salt (WST)-8 was used to measure the cellular viability. Flow cytometry was used to measure the active oxygen and apoptosis index. The expression of nuclear factor erythroid 2-related factor 2 (Nrf2), hemeoxygenase-1 (HO-1), Bcl-2 and Bax protein were measured by Western blot. Results Compared with group A, the WST-8 showed that the viability of Müller cells apparently decreased in group B (t=39.59,P＜0.05). Compared with the group B, the viability of Müller cells had changes in group C (t=0.97,P＞0.05), but recovered in group D and E (t=−4.17, −7.52;P＜0.05). Compared with group A, the FCM showed that the mitochondrial ROS levels was higher in group B (t=−30.99,P＜0.05). Compared with group B, the mitochondrial ROS levels were decreased in group D (t=27.68,P＜0.05). Compared with group A, Bax, Nrf2 and HO-1 increased (t=–11.03, –63.17, –11.44;P＜0.05), while the bcl-2 decreased in group B (t=7.861,P＜0.05). Compared with the group B, Nrf2, HO-1 and Bax decreased (t=15.11, 26.59, 6.27;P＜0.05), while the bcl-2 increased in group D (t=−6.53,P＜0.05). Conclusions Under the high glucose, CPDT may reduce the mitochondrial ROS levels and the expression of Nrf2, HO-1 and Bax protein of Müller cells. It may inhibit apoptosis through activating the Nrf2/HO-1 pathway and balancing of level of Bcl-2 protein and mitochondrial ROS.
ObjectiveTo explore and analyze the nutritional risk and dietary intake of patients with coronavirus disease 2019 (COVID-19), and provide data support for nutritional intervention.MethodsCOVID-19 inpatients were investigated in Wuhan Wuchang Hospital and the People’s Hospital of Wuhan University (East Area) from March 9th to 16th, 2020 by Nutrition Risk Screening 2002 (NRS 2002) scale and designed questionnaire. The energy and protein requirements were calculated according to the standard of 30 kcal/(kg·d) and 1.2 g/(kg·d). The nutritional risk, energy and protein intake, body weight and body mass index and their changes in the mild and severe patients were analyzed. The energy and protein intake of the two types of nutritional risk patients was analyzed.ResultsA total of 98 patients with COVID-19 completed the investigation, in whom 46 (46.94%) had nutritional risk, including 32 (39.02%) with mild type and 14 (87.50%) with severe type; and the difference was statistically significant (P<0.001). Compared with the usual condition, the body weight and body mass index of the two types of patients significantly decreased (P<0.01 or P<0.001); the energy and protein intake in mild type patients were significantly higher than those in the severe type patients (P<0.001); compared with the requirement, the protein intake in the two types of patients were significantly lower than the demand, while the energy and protein intake in the mild type patients were significantly lower than the requirement (P<0.05 or P<0.01). The proportion of energy and protein intakes in patients with nutritional risk was significantly higher than that in patients without nutritional risk (P<0.001 or P<0.01); the energy and protein intakes in patients without nutritional risk was significantly higher than that in patients with nutritional risk (P<0.001); the protein intakes in patients with nutritional risk was obviously insufficient (P<0.001); while the energy intake of the patients without nutritional risk was higher than the requirement (P<0.001).ConclusionsCOVID-19 patients has high incidence of nutritional risk which was higher in the severe patients compared with the mild patients. Higher incidence and lower intake of energy and protein are in the severe patients compared with those in the mild patients. Patients with nutritional risk has a higher proportion of energy and protein inadequate intake and lower intake compared with the patients without nutritional risk.
Protein post-translational modifications (PTMs) are critical for modulating protein structure and function. Among these, lysine lactylation (Kla) has garnered significant attention in recent years as a newly discovered PTM. Although Kla has been thoroughly investigated in eukaryotic systems, its study in prokaryotes, especially bacteria, remains comparatively limited. Emerging research highlights that bacterial Kla, operating through dynamic modification mechanisms, is pivotal in processes such as growth and metabolism, virulence control, pathogenicity, and host-pathogen interactions. This article provides a comprehensive overview of the latest progress in bacterial Kla research, emphasizing its historical discovery, distinct modification features, and underlying molecular regulatory mechanisms. We further explore the regulatory roles of this modification in bacterial physiological processes and pathogenesis, concluding with a discussion of current research challenges and prospective future developments.
Objective To explore the expression of survivin gene in retinoblastoma (RB) and its relationship with the stages and histodifferentiation degree of RB and the expression of p53、bcl-2 proteins. Methods Expression of survivin, p53 and bcl-2 proteins in 38 RB conventional paraffin samples were detected with survivin, p53 and bcl-2 monoclonal antibodies respectively by immunohistochemical assay. The expression of survivin of normal retina in 6 control samples was observed. Results In 38 cases of RB, positive expression of survivin was found in 20 (52.6%); while none of the 6 normal retinal tissue expressed survivin, which had significant difference between the two group (P＜0.05). The positive expression of survivin did not correlate with sex of patient, disease stages and histological type (P＞0.05). In 38 RB cases, positive expression of p53 was in 25 with the rate of 65.8%, and of bcl-2 in 18 with the rate of 47.4%. The positive-expressed rates were much higher in positive-expressed p53 and bcl-2 group than those in the negative-expressed p53 and bcl-2 group(P＜0.05). Conclusion The increase of the expression of survivin implies that it may take part in the occurrence and development of RB; the interaction among survivin, p53 and bcl-2 may participate in the access and the course of RB. (Chin J Ocul Fundus Dis，2004，20：215-217)
Objective To investigate the role of ephrin A genes in the development of oxygen induced retinalneovascularization (OIR) in mice.Methods The OIR model was established by oxygen induction in new born C57BL/6J mice.Reversed transcript polymerase chain reaction (RT-PCR) was used to measure the expression levels of ephrin A1-A5 in retinas of mice in experimental and normal control group.Results All of the ephrin A family genes expressed in normal retinas. Ephrin A1 mRNA was significantly higher in OIR group(t=3.19,P=0.019); ephrin A2 mRNA was higher in the 15-day-old OIR retinas(t=3.71,P=0.033）; ephrin A3-A5 mRNA decreased or disappeared in 12 and 13-day-old RNV mice, and increased in 15-day-old OIR mice. Conclusion Ephrin A genes are involved in the development of retina and OIR.
The fundus lesions caused by high myopia (HM) often lead to irreversible visual impairment or even blindness. However, the pathogenesis of HM and its fundus lesions is still unclear, the intraocular fluid detection technology of micro samples has brought new prospects for the early diagnosis, monitoring and intervention of the fundus lesions. The molecules associated with HM are various and functionally diverse, intermolecular interactions are staggered and the specific mechanism is complex. With the development of intraocular fluid detection technology, while gradually revealing the role of each molecule in the pathogenesis of HM, it is expected to successfully assist clinical work in the future, providing outpost markers for the progress of myopia and targets for early intervention, or providing a new therapy choice for HM fundus lesions at the molecular level targeting pathogenesis, which is expected to provide more accurate and effective treatment for HM patients in the future.
ObjectiveTo investigate key differentially expressed genes (DEGs) in peripheral blood of idiopathic epilepsy patients, as well as their biological functions, cellular localization, involved signaling pathways, through bioinformatics analysis. So to provide new insights for the pathogenesis and prevention of idiopathic epilepsy.MethodsFirstly, we screened and downloaded microarray data including 6 peripheral blood samples of drug-naive patients with idiopathic epilepsy, 8 peripheral blood samples of responders of idiopathic epilepsy treated with Valproate (VPA), and 10 peripheral blood samples of non-responders of idiopathic epilepsy treated with VPA from Gene Expression Omnibus (GEO) data series GSE143272, which Public in January 2020. Secondly, we identified DEGs via the limma package and others in R software. Then we had gotten 74 DEGs, and subsequently conducted gene ontology and pathway enrichment analysis, PPI network analysis and hub gene analysis, using multiple methods containing DAVID, STRING, and Cytohubba in Cytoscape.ResultsWe had identified significant hub DEGs, including TREML3P, KCNJ15, ORM1, RNA28S5, ELANE, RETN, ARG1, LCN2, SLPI, HP, PGLYRP1, BPI, DEFA4, TCN1, MPO, MMP9, CTSG, CXCL8, RNASE3, RNASE2, S100A12, DEFA1B, DEFA1, DEFA3, CEACAM8, MS4A3, PTGS2, PI3, CCL3. The biological processes involved in these DEGs include immune response, inflammatory response, chemotaxis, etc. While, the molecular function is focused on peroxidase activity, chemokine activity, etc. Moreover, KEGG pathway enrichment analysis shows that DEGs were mainly involved in cytokine-cytokine receptor interaction, Toll-like receptor signaling pathway, chemokine signaling pathway and so on.ConclusionThese important key DEGs may be involved in the onset and development of idiopathic epilepsy through a variety of signaling pathways and complex mechanisms.
Objective To clone the genes of nogo-66 and NEP1-40 from spinal cord of rat and to realize the expression of its protein in vitro. Methods The nogo-66 and NEP1-40 genes were cloned from the spinal cord of juvenil rat by use of RT-PCR techniques, and the objective genes were bonded to T vector through gene coupled action, recombinant plasmid were sequencing, and the genes were cloned into PQE30-GST vector, then the recombinant plasmids were induced by isopropylthiogalactoside(IPTG) to express the proteins. The two proteins were purified by Ni-column and detected by using Westernblot test. Results The Nogo-66 and NEP1-40 genes were successfully cloned from rat, which were 215 bp and 137 bp for each one when add the enzyme site. No gene mutations were detected in the two genes after sequencing. The expression plasmids were cut by the two enzyme (BamH Ⅰ and Hind Ⅲ), the target bands were seen on the results of electrophoresis. The expression plasmids were induced by IPTG and got the purified GST fusion protein nogo-66 and NEP1-40, which relative molecular weight were 33.2×103 and 30.3×103 respectively. The results of Westernblot test confirmed that the antigenicity of the two proteins was precise. Conclusion Nogo-66 and NEP1-40 proteins can be expressed in a high efficiency in vitro using genetic engineering, so it provides a good basis for further research on its function and vaccine for spinal injury.