1.Mechanism of circ_0038467 regulating oxygen-glucose deprivation-induced nerve cell damage by targeting miR-940
Xuan-Dong KONG ; Li-Qin ZHOU ; Ning WANG ; Tian-Ya WU ; Ming ZHAO
Chinese Pharmacological Bulletin 2024;40(5):887-893
Aim To explore the effect of circ_0038467 on nerve cell damage induced by hypoxia-glucose dep-rivation(OGD)and its possible mechanism.Methods Rat cortical nerve cells were isolated and cultured,and then induced by OGD to establish a cell injury model.si-NC,si-circ_0038467,miR-NC,and miR-940 mimics were transfected into rat cortical nerve cells and treated with OGD for 6 h.si-circ_0038467 and an-ti-miR-NC or anti-miR-940 were co-transfected into rat cortical neurons,followed by OGD treatment for 6 h.qRT-PCR was used to detect the expression levels of circ_0038467 and miR-940.CCK-8 method and flow cytometry were used to examine cell proliferation and apoptosis.LDH method was used to detect cell dam-age.The dual luciferase reporter experiment was used to detect the targeting relationship between circ_0038467 and miR-940.Western blot was employed to detect cleaved caspase-3 and cleaved caspase-9 protein levels.Results Circ_0038467 expression increased and miR-940 expression decreased in OGD-induced nerve cells(P<0.01).After transfection with si-circ_0038467 or miR-940 mimics,cell survival rate in-creased(P<0.01),while LDH release rate,apopto-sis rate,and the protein levels of cleaved caspase-3 and cleaved caspase-9 decreased(P<0.01).Circ_0038467 could target miR-940.Compared with the OGD+si-circ_0038467+anti-miR-NC group,cell survival rate in OGD+si-circ_0038467+anti-miR-940 group was down-regulated(P<0.01),while LDH re-lease rate,apoptosis rate and cleaved caspase-3,cleaved caspase-9 levels were up-regulated(P<0.01).Conclusion Interference of circ_003 8467 and could protect nerve cells from OGD-induced oxidative stress and apoptosis by up-regulating miR-940.
2.Epidemiological and molecular characteristics of human metapneumovirus in children in Tianjin
Rui LI ; Liru GUO ; Mei KONG ; Ming ZOU ; Xiaochang LIU ; Tao WANG ; Mengxue GAO ; Xiaoyan LI
Chinese Journal of Microbiology and Immunology 2024;44(7):588-595
Objective:To investigate the epidemiological and genetic characteristics of human metapneumovirus (hMPV) in Tianjin during two influenza epidemic seasons from October 2020 to March 2021 and from October 2021 to March 2022, and enrich the whole genome database of hMPV in China.Methods:A total of 1 040 pharyngeal swab samples collected from patients with influenza-like illness (ILI) were analyzed using microfluidic chip fluorescence quantitative PCR. RT-PCR was used to amplify the whole genome in hMPV-positive samples, and the second-generation sequencing was performed for complete genome sequencing. Bioinformatics software including CLC, DNAStar, and MEGA was used for sequence assembly, nucleotide and amino acid homology analysis, and phylogenetic tree mapping.Results:Among the 1 040 samples, 25 were positive for hMPV with a positive rate of 2.40%. The highest positive rate was observed in the age group of 3 to 5 years, reaching 3.71% (16/431). During the influenza epidemic seasons, the detection rate of hMPV peaked in December, reaching 6.67% (12/180). Twelve strains were successfully sequenced, and there were seven of type B2, four of type A2b, and one of type B1. More variations were detected in the G gene, with 111nt-dup sequence repeats observed in the G gene of three A2b strains.Conclusions:The prevalence of hMPV peaks in December during the influenza epidemic seasons in Tianjin, with Type B2 being the predominant type. Except for the G gene with more mutations, other genes remain stable.
3.Effect of matrine on diabetic nephropathy mices based on the TGF-β/Smads signaling pathway
Jing KONG ; You-Li HE ; Ming-Ming YANG ; Ying LIU
The Chinese Journal of Clinical Pharmacology 2024;40(13):1923-1927
Objective To investigate the effect of matrine on improving diabetic nephropathy(DN)through transforming growth factor β/signal transduction protein(TGF-β/Smads)pathway.Methods C57BL/6 mice were randomly divided into NC group,DN group,DN+matrine group,and DN+metformin group,with 10 mice in each group.Except for NC group,other groups were established DN models by high-fat and high-sugar diet combined with intraperitoneal injection of streptozotocin(STZ).After successful modeling,DN+matrine group was treated with matrine for 4 weeks,DN+metformin group was treated with metformin,DN and NC groups were given equal volumes of 0.9%NaCl.Western blot was used to detect the expression of collagen Ⅳ(Col Ⅳ),fibronectin(FN),Smad family member 2/3(Smad2/3),and phosphorylated Smad2/3(p-Smad2/3)in kidney.Biochemical method were used to detect blood glucose,creatinine,and urea nitrogen levels of mice.Results In NC group,DN group and DN+matrine group,blood glucose levels were(8.16±2.53),(24.84±4.67)and(23.88±3.57)mmol·L-1;blood creatinine levels were(36.48±5.63),(97.51±10.59)and(41.88±7.26)mmol·L-1;urea nitrogen levels were(53.11±4.72),(91.50±8.62)and(63.29±5.69)mmol·L-1;Col Ⅳ protein expression levels in kidney were 1.00±0.19,3.34±0.58 and 1.99±0.44;FN protein expression levels were 1.00±0.21,3.63±0.47 and 1.79±0.43;p-Smad2/3/Smad2/3 were 1.00±0.18,2.74±0.51 and 1.08±0.33.Above indicators showed statistically significant differences between NC group and DN group(all P<0.05).Conclusion Matrine can effectively inhibit the activation of TGF-β1/Smads signaling pathway and reduce the polyproduct of extracellular matrix-related proteins,thus playing a protective role in DN kidney injury.
4.Comparison of Methods between Soiled Bedding Sentinels and Exhaust Air Dust PCR for Health Monitoring of Rodent Laboratory Animals
Lingzhi YU ; Xiaofeng WEI ; Ming LI ; Zhihao KONG
Laboratory Animal and Comparative Medicine 2024;44(3):321-327
The microbiological quality of laboratory animals is crucial for the validity and reproducibility of scientific research data,as well as human health and animal welfare.Currently,individual ventilation cages(IVC)have become the mainstream feeding system for rodent laboratory animals.The most commonly used pathogen monitoring method for this feeding system is soiled bedding sentinels(SBS).This method monitors the microbial carrying status of mouse colony through indirect contact and delayed feedback.It can effectively monitor pathogens transmitted via the fecal-oral route,such as mouse hepatitis virus and reovirus.However,this method has difficulty detecting pathogens mainly transmitted through aerosols or direct contact,such as Sendai virus and Pasteurella pneumotropica.The exhaust air dust(EAD)-PCR monitoring method involves swab sampling in the IVC exhaust ducts to monitor the corresponding racks of the ducts;swab sampling before the prefiltration of the host to monitor the entire IVC rack;and EAD collection device sampling to monitor all racks connected to the same host.Different IVC manufacturers have developed corresponding EAD collection devices for their respective IVC systems,making operations convenient and standardization easy.Compared with the SBS method,the EAD-PCR method significantly improves detection rate and timeliness,with the fastest detection possible after one week of exposure.It can serve as a supplement or replacement for the SBS method.Currently,increasing evidence supports that EAD-PCR testing is a more reliable,sensitive,and cost-effective monitoring method,and is more beneficial to animal welfare.This article reviews the application progress of these two methods for monitoring pathogens,analyzes the existing limitations of the EAD-PCR method,and proposes solutions based on its implementation in our laboratory and examination units.The EAD-PCR method helps reduce the number of live sentinel animals used in pathogen monitoring,in order to better maintain the"3Rs"principle of laboratory animal welfare.
5.Analysis on influencing factors of severity in patients with type 2 diabetes mellitus complicating cerebral small vessel disease
Juan JI ; Ming LI ; Tianchen WU ; Xiaodan SUN ; Xuesu WU ; Wenwen KONG
Chongqing Medicine 2024;53(15):2277-2283
Objective To screen and analyze the influencing factors of severity in the patients with type 2 diabetes mellitus (T2DM) complicating cerebral small vessel disease (CSVD).Methods A total of 519 pa-tients with T2DM complicating CSVD admitted and treated in Nanjing Municipal Hospital of Traditional Chi-nese Medicine from June 2018 to May 2023 were selected and divided into the mild group (n=214) and the se-vere group (n=305) according to the CSVD imageological score.The relevant demographic,laboratory and imageological indicators were collected.The influencing factors of T2DM complicating CSVD were screened out by the LASSO and Logistic regression analysis and the predictive model was established.The receiver op-erating characteristic (ROC) curve,goodness of fit evaluation and restricted cubic spline (RCS) fitting curve were drawn to analyze the dose-response relationship between Cys C,albumin/globulin (A/G) ratio with the disease severity.Results The male proportion and age in the severe group were greater than those in the mild group,neutrophil,systemic immune-inflammation index (SII),creatinine (Crea),uric acid (UA),Urea (Ure-a),D-dimer (D-D),lactate dehydrogenase (LDH),adenosine deaminase (ADA),globulin (GLB) and Cys C were higher than those in the mild group,lymphocyte,ALT,High density Lipoprotein-cholesterol (HDL-C),serum cholinesterase (CHE),prealbumin (PAB),and A/G were lower than those in the mild group,and the differences were statistically significant (P<0.05).LASSO and logistic regression analysis showed that the gender,age,A/G and Cys C were the independent influencing factors in the patients with T2DM complicating CSVD.The area under the curve (AUC) of this model was 0.658 (95%CI:0.610-0.706) with goodness of fit (P=0.520).The RCS fitting curves showed that serum Cys C≥0.618 mg/L had a linear relationship with CSVD imageological score (P=0.035),and A/G≥1.268 had a nonlinear relationship with CSVD imageologi-cal score (P=0.007).Conclusion The advanced age,male,increased Cys C level and decreased A/G in the pa-tients with T2DM complicating CSVD are the independent risk factors for the severity of whole brain damage.
6.Analysis of characteristic factors and establishment of diagnostic model for hyperuricemia caused renal impairment
Xiaodan SUN ; Tianchen WU ; Ming LI ; Juan JI ; Xuesu WU ; Wenwen KONG
Chongqing Medicine 2024;53(18):2750-2754,2760
Objective To screen the characteristic factors of renal impairment occurrence in the patients with hyperuricemia,and to analyze its diagnostic value and establish a diagnostic model.Methods A total of 2405 inpatients with diagnosed hyperuricemia in the Nanjing Municipal Hospital of Traditional Chinese Medi-cine,Nanjing University of Traditional Chinese Medicine from December 2018 to December 2022 were selected and divided into the kidney lesion group (n=1343) and the non-kidney lesion group (n=1062) according to eGFR.The characteristic factors of hyperuricemia caused renal impairment were screened and analyzed by Lasso and logistic regression and the diagnostic model was constructed.The diagnostic value of characteristic factors and diagnostic model were evaluated by the receiver operating characteristic (ROC) curve,and the change rule between the characteristic factors and the results was found by the restricted cubic splines (RCS) fitting.Results The age,uric acid (UA),cystatin-C (Cys-C) and retinol-binding protein (RBP) were the characteristic factors of hyperuricemia caused renal impairment.The combined diagnostic model:logit (P)=-8.70+0.602×age (10 years old)+0.033×UA (10 μmol/L)+0.277×Cys-C (0.1 mg/L)+0.189×RBP (10 mg/L),the area under the ROC curve (AUC) of the combined diagnosis model was 0.893 (95%CI:0.880-0.905).For every 10 μmol/L increase in blood UA,the risk of renal impairment occurrence in hyperurice-mia was increased by 3%;for every 10 years increase in age,the risk of renal impairment occurrence in hyperu-ricemia was increased by 83%;for every 10 mg/L increase in RBP,the risk of kidney damage occurrence of re-nal impairment in hyperuricemia was increased by 21%;for every 0.1 mg/L increase in Cys-C,the risk of re-nal impairment occurrence in hyperuricemia was increased by 32%.Conclusion The combined diagnostic model for whether the renal impairment in the patients with hyperuricemia occurring has good diagnostic val-ue,and Cys-C deserves more attention.
7.Classification Method for Petroleum Pollutants Based on Inception-One-Dimensional Convolutional Neural Network and Infrared Spectroscopy
De-Ming KONG ; Shao-Wei HE ; Xin-Yi LI ; Jun-Yu ZHAO ; Xiao-Dong NING
Chinese Journal of Analytical Chemistry 2024;52(9):1287-1297
Infrared spectroscopy technology has many advantages such as high efficiency and non-destructiveness,and has an important research and application value in the field of petroleum pollutant classification and detection.In this study,a petroleum pollutant classification method by combing the discrete wavelet transform(DWT)algorithm and a one-dimensional convolutional neural network based on the Inception module(Inception-1D-CNN)was proposed.Firstly,the DWT algorithm was used to denoise the original infrared spectral data to eliminate the interference information caused by experimental environment,instrument error and manual operation.Then,the inception-1D-CNN model was used to obtain multi-scale infrared spectroscopy feature information,and then classify the petroleum pollutants.Experimental results showed that compared with preprocessing methods such as standard normal variable(SNV),adaptive iteratively reweighted penalized least squares(AirPLS),and Savitzky-Golay smoothing(S-G),the prediction accuracy of the DWT algorithm combined with the 1D-CNN model with a convolutional kernel size of 3×1 was 86.6%,which was 6.6%,6.6%and 3.3%higher,respectively.The prediction accuracy of DWT algorithm combined with 1D-CNN model with a convolutional kernel size of 5×1 was 93.3%,which was 10.0%,7.0%and 3.3%higher,respectively.The prediction accuracy of the DWT algorithm combined with the 1D-CNN model with a convolutional kernel size of 7×1 was 90.0%,which was 6.7%,10.0%and 3.4%higher,respectively.The prediction accuracy of the DWT algorithm combined with the inception-1D-CNN model was 100.0%,which was 10.0%,10.0%and 3.4%higher,respectively.Therefore,the DWT algorithm combined with the inception-1D-CNN model could accurately classify and predict petroleum pollutants,and provided a certain basis for the subsequent treatment of oil spills on the sea surface.
8.Molecular study of a case with variant of RHCE*ce allele in haplotype dce resulting in weakened e antigen
Yongkui KONG ; Hecai YANG ; Ming SHAO ; Yinghui CHEN-LI ; Wanjin ZHANG ; Xiaoyan ZHANG ; Jing WANG ; Xianping LYU ; Qiankun YANG
Chinese Journal of Medical Genetics 2024;41(9):1039-1044
Objective:To explore the RH genotype for a female with RhD(-) blood type and its molecular basis. Methods:A 26-year-old female who had attended the outpatient clinic of the First Affiliated Hospital of Zhengzhou University in August 2019 was selected as the study subject. Peripheral blood samples were collected from the proband and her parents for Rh phenotyping with gel card method. PCR-sequence-based typing (PCR-SBT) and DNA sequencing were used to determine the RHD zygosity and RH genotype of the proband and her parents. Homology modeling of Rh proteins was performed with bioinformatic software, and protein structural alterations caused by the variant was simulated by molecular dynamics. This study was approved by the Medical Ethics Committee of the First Affiliated Hospital of Zhengzhou University (Ethics No. 2023-KY-0870-003). Results:Serological tests showed that the proband and her father both had weakened e antigen of the Rh phenotype. PCR-SBT and DNA sequencing showed that the genotypes of the proband and her parents were dce/ dCE, dce/ DcE and dCE/ DcE, respectively. And the genotypes of the RHD and RHCE of the proband were RHD*01N.01/ RHD*01N.16, RHCE*01.01/RHCE*04, respectively. Protein simulation and molecular dynamics analysis revealed that the ce_16C variant resulted from RHCE* ce (c.48G>C) may alter the structure of intracellular and extracellular loops, mainly affecting the mobility of extracellular loops 2, 6 and intracellular loops 3, 4. Conclusion:Variant of the RHCE* ce allele c. 48G>C probably underlay the weakened e antigen in this proband.
9.Comparative analysis of handheld full process visual navigation and robot assisted total hip arthroplasty results
Yi LI ; Peng REN ; Guoqiang ZHANG ; Xin ZHI ; Ming NI ; Xiangpeng KONG ; Wang GU
Chinese Journal of Orthopaedics 2024;44(21):1393-1400
Objective:To compare the precision and stability of handheld full-course visual navigation and robot-assisted total hip arthroplasty (THA), as well as to evaluate perioperative laboratory indicators and clinical efficacy.Methods:A retrospective analysis was performed on 68 patients with unilateral hip joint disorders who underwent primary THA at the General Hospital of the People's Liberation Army of China from January to June 2024. Patients were divided into two groups based on the assistance method: the Visual Treatment Solution (VTS) group, which included 34 patients (13 males, 21 females; mean age 56.92±8.31 years; body mass index[BMI] 24.20±3.55 kg/m 2), and the MAKO group, also with 34 patients (18 males, 16 females; mean age 57.97±6.54 years; BMI 25.20±3.91 kg/m 2). Among the VTS group, 1 there were 18 cases on the left side and 16 cases on the right side, including 14 cases of femoral head necrosis, 8 cases of congenital hip dysplasia, and 12 cases of hip osteoarthritis. In the MAKO group, there were 19 cases on the left side and 15 cases on the right side, including 13 cases of femoral head necrosis, 10 cases of congenital hip dysplasia, and 11 cases of hip osteoarthritis. Preoperative planning, intraoperative verification deviations, postoperative acetabular prosthesis anteversion and abduction angles, and limb length discrepancies were analyzed. Acetabular angle distribution scatter plots were generated for intraoperative verification and postoperative imaging. The operation time, hospital stay, and laboratory values, including hemoglobin (Hb) loss, fibrinogen degradation products (FDP), D-dimer, high-sensitivity C-reactive protein (hCRP), as well as postoperative pain (measured by the visual analogue scale, VAS), hip Harris score (HHS), and Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) scores were recorded. Results:No statistically significant differences were observed between postoperative imaging measurements of anteversion and abduction angles and intraoperative verification values ( P>0.05). Postoperative imaging indicated that 91% (31/34) of the VTS group and 100% (34/34) of the MAKO group were within the Lewinnek safety zone. The scatter distribution of angles in the MAKO group was more concentrated than that in the VTS group. Furthermore, the angle deviation between intraoperative verification and preoperative planning was significantly greater in the VTS group for both anteversion (2.00° [-1.00°, 4.00°] vs. 0.00° [-1.00°, 1.00°]) and abduction angles (-1.00° [-5.00°, 0.00°] vs. 0.00° [-1.00°, 1.00°]; P<0.05). The bilateral limb length discrepancies were 1.35(0.67, 2.45) mm in the VTS group and 1.65(0.50, 2.52) mm in the MAKO group, with no significant difference noted ( P>0.05). Both methods achieved effective hip biomechanical reconstruction. Additionally, no significant differences in operation time, Hb loss, FDP, D-dimer, hCRP, or postoperative hospital stay were found ( P>0.05). Follow-up at one and three months postoperatively showed no significant differences in VAS (2.94±0.92 vs. 2.97±0.83), HHS (69.88±4.20 vs. 70.06±3.88), or WOMAC scores (22.44±3.15 vs. 22.76±3.39) between the two groups ( P>0.05). Conclusion:The stability of preoperative planning and design in robot-assisted total hip arthroplasty is superior to that of handheld navigation methods. Nevertheless, both approaches demonstrate comparable postoperative prosthesis stability, laboratory indicators, and clinical outcomes.
10.Establishment of a method for determining the key parameters of hydrolysis kinetics of acid α -glucosidase for injection by ion chromatography
Xin-yue HU ; Jia-hao KONG ; Yue SUN ; Lü-yin WANG ; Xiao-ming ZHANG ; Ping LÜ ; Cheng-gang LIANG ; Jing LI
Acta Pharmaceutica Sinica 2024;59(12):3361-3366
The Dionex CaboPacTM PA10 BioLCTM Analyical 2 mm × 250 mm column was used with a protective column (Dionex CaboPacTM PA10 BioLCTM Guard 2 mm × 50 mm). 100 mmol·L-1 sodium hydroxide solution was used as eluent; the flow rate was 0.25 mL·min-1. Sample tray temperature: 35 ℃. The pulse amperometric detector was adopted, and the waveform was Gold CWE, Ag-AgCl RE, Carbo, Quad. The samples were cultured with 8 concentrations of glycogen substrates (0.31, 1.25, 2.5, 5, 10, 20, 30, and 40 mg·mL-1).

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