Objective:To demonstrate any role of iron metabolism in the inhibition by aerobic exercise of myocardial apoptosis in atherosclerotic mice.Methods:Eight-week-old male ApoE -/- gene knockout mice were randomly divided into a control group, a model group and an aerobic exercise group, each of 9. A model of atherosclerosis was induced in the rats of the model and aerobic exercise groups by feeding them a " western" diet for 12 weeks. During that time the aerobic exercise group only was given aerobic exercise training. The control group was fed normal rat chow during that period. Myocardial apoptosis was detected using TUNEL staining, and the expression and localization of ferritin heavy chain 1 (FTH1) and glutathione peroxidase 4 (GPX4) in the myocardium used immunohistochemistry. Western blotting was applied to detect the FTH1 and GPX4 protein levels, and iron deposition in the myocardium was detected using Prussian blue staining. Iron, lipid peroxide malondialdehyde (MDA) and glutathione peroxidase (GSH-PX) in the myocardial tissue were also measured. Results:The TUNEL staining showed significant apoptosis in the model group. In the aerobic exercise group it was significantly less. There was obvious iron deposition in the myocardia of the model group, which was significantly reduced in the aerobic exercise group. The average FTH1 and GPX4 levels in the model group were lower than in the control group, and significantly elevated in the aerobic exercise group.in the aerobic exercise group than in the model group. Iron and MDA levels in the aerobic exercise group were significantly lower, on average, than among the model group, while that of GSH-PX was significantly higher.Conclusions:Aerobic exercise can significantly inhibit cardiomyocyte apoptosis in atherosclerotic mice. The mechanism may be closely related to better iron metabolism, reduced oxidative stress and the inhibition of iron overload.

ObjectiveTo understand the current research status, research hotspots and development trends of constraint-induced movement therapy (CIMT) in the field of rehabilitation. MethodsThe relevant articles of CIMT in rehabilitation from January, 2000 to October, 2022 in CNKI and Web of Science were retrieved. The authors, institutions, countries, keywords and burst words were extracted with CiteSpace 6.1.R3 to draw knowledge mapping. ResultsA total of 1 165 articles were included, 359 articles in Chinese and 806 in English. The trend of annual publications was generally consistent, and after a period of rapid growth, the current annual publications showed a fluctuating trend. There was more cooperation among the institutions in English articles, with geographical limitation. The institutions in Chinese articles had the problem of insufficient cooperation. The researches mainly focused on the application of CIMT in different diseases, the improvement of motor function by CIMT, the application of CIMT in combination with other therapies, and the study of the related mechanisms of CIMT. In recent years, Chinese burst keywords included modified constraint-induced movement therapy, stroke hemiparesis, clinical efficacy and repetitive transcranial magnetic stimulation; English burst keywords included transcranial direct current stimulation, non-invasive brain stimulation, and unilateral cerebral palsy. ConclusionResearch on CIMT in the field of rehabilitation is in a period of steady development, and CIMT combined with non-invasive brain stimulation is likely to be a hotspot in future research.

OBJECTIVE: To analyze variants of SMN gene in a Chinese pedigree affected with Spinal muscular atrophy (SMA). METHODS: A Chinese pedigree diagnosed at the Nanchang First Hospital in January 2020 was selected as the study subject. Peripheral blood samples were collected for the extraction of DNA. All exons of the SMN gene were detected by multiple ligation-dependent probe amplification (MLPA). Potential variants of the SMN gene were also detected by Whole exome sequencing (WES), and the result was verified by Sanger sequencing. cDNA extracted from fresh blood sample was used as a template to verify the location of variant on the SMN genes. RESULTS: The proband was found to harbor a heterozygous deletion of the SMN1 Exon7+Exon8, and a heterozygous c.81G>A variant. The SMN1 Exon7+Exon8 deletion was inherited from her father and grandmother, whilst the c.81G>A variant was inherited from her mother and maternal grandfather. Her aunt was also a carrier of the heterozygous deletion, while her paternal aunt, her husband, and their daughter were not. cDNA amplification and Sanger sequencing confirmed that the c.81G>A variant was located in the SMN1 gene. CONCLUSION MLPA combined with NGS and Sanger sequencing can identify compound heterozygous variants of the SMN gene in the SMA patients.
Female ; Humans ; Male ; DNA, Complementary ; East Asian People ; Fathers ; Mothers ; Muscular Atrophy, Spinal/diagnosis* ; Pedigree ; Survival of Motor Neuron 1 Protein/genetics*

Objective:To evaluate the role of histone deacetylase 6 (HDAC6) in spinal dorsal horn in dexmedetomidine-induced reduction of neuropathic pain (NP) in rats.Methods:Forty clean-grade healthy male Sprague-Dawley rats, aged 7-9 weeks, weighing 190-240 g, were divided into 5 groups ( n=8 each) using a random number table method: control group (group C), sham operation group (group SH), group NP, dexmedetomidine group (group D), and specific HDAC6 inhibitor ACY-1215 plus dexmedetomidine group (group AD). The animals were commonly fed without any treatment in group C. The sciatic nerve was only isolated but not ligated in group SH.The animals were anesthetized with intraperitoneal 10% chloral hydrate 350 mg/kg.NP was induced by chronic constrictive injury (CCI). The right sciatic nerve was exposed, and 4 loose ligatures were placed on the sciatic nerve at 1-mm intervals with 4-0 silk thread in NP and D groups.In group D, dexmedetomidine 40 μg/kg was intraperitoneally injected once a day starting from the end of operation until the animals were sacrificed.In group AD, ACY-1215 25 mg/kg was intraperitoneally injected every day immediately before CCI, and dexmedetomidine 40 μg/kg was intraperitoneally injected daily after CCI until 15 days after CCI.The equal volume of solvent was given instead of dexmedetomidine in S and NP groups.The mechanical paw withdrawal threshold to von Frey filament stimulation (MWT) and thermal paw withdrawal latency (TWL) were measured at 1 day before CCI (baseline, T 0) and 3, 6, 9, 12 and 15 days after CCI (T 1-5). The rats were then sacrificed, and the dorsal horn tissues of L 4-6 spinal cord were obtained for determination of the expression of myeloid differentiation factor 88 (MyD88) and nuclear factor kappa B (NF-κB) p65 by Western blot. Results:Compared with group C and group SH, the MWT was significantly decreased, and the TWL was shortened at T 1-5, and the expression of MyD88 and NF-κB p65 in the spinal dorsal horn was up-regulated in NP, D and AD groups ( P<0.05). Compared with group NP, the MWT was significantly increased, and the TWL was prolonged at T 1-5, the expression of MyD88 and NF-κB p65 in the spinal dorsal horn was down-regulated in group D ( P<0.05), and no significant change was found in the parameters mentioned above in group AD ( P>0.05). Compared with group D, MWT was significantly decreased, and TWL was shortened at T 1-5, and the expression of MyD88 and NF-κB p65 was up-regulated in the dorsal horn of the spinal cord in group AD ( P<0.05). Conclusion:HDAC6 in spinal dorsal horn is involved in dexmedetomidine-induced reduction of NP in rats, which is related to inhibiting MyD88/NF-κB signaling pathway.

Objective: To evaluate the effect of penehyclidine hydrochloride (PHC) on Toll-like receptor 4 (TLR4)/nuclear factor kappa B (NF-κB) signaling pathway in non-ventilated lung injury in the patients undergoing radical operation for lung cancer. Methods: A total of 100 patients, aged 40-64 yr, with body mass index 18-27 kg/m², of American Society of Anesthesiology physical status Ⅱ or Ⅲ, undergoing radical operation for lung cancer, were divided into 2 groups (n=50 each) according to the random number table method: control group (group C) and PHC group.PHC 0.01 mg/kg was intravenously injected at 10 min before anesthesia induction in group PHC, while the equal volume of normal saline was given instead in group C. The peripheral tissues of the removed lung tissues were obtained for determination of wet/dry weight ratio (W/D ratio). The pathological changes and ultrastructure of lung tissues were observed under light microscope, and lung injury was assessed and scored.The expression of TLR4 and NF-κB protein and mRNA was detected by Western blot and real-time polymerase chain reaction, respectively.Before administration (T₀), at the onset of one-lung ventilation (T₁), at 60 min of one-lung ventilation (T₂), immediately after the end of one-lung ventilation (T₃), at the end of operation (T₄) and at 24 h after operation (T₅), blood samples were collected from the internal jugular vein for determination of serum tumor necrosis factor-alpha, interleukin-6 (IL-6) and IL-8 concentrations by enzyme-linked immunosorbent assay. Results: Compared with group C, the W/D ratio and lung injury scores were significantly decreased, the expression of TLR4 and NF-κВ protein and mRNA was down-regulated, and the concentrations of tumor necrosis factor-alpha, IL-6 and IL-8 were decreased at T₂-T₅ in group PHC (P<0.05). The pathological changes and damage to ultrastructure of lung tissues were significantly attenuated in group PHC as compared with group C. Conclusion The mechanism by which PHC attenuates non-ventilated lung injury is related to blocking TLR4/NF-κВ signaling pathway and reducing inflammatory responses in the patients undergoing radical operation for lung cancer.

The effect of methanol addition on the heterologous expression of isoprenyl transferase NovQ was studied in Pichia pastoris Gpn12, with menadione and isopentenol as precursors to catalyze vitamin K2 (MK-3) synthesis. The expression of NovQ increased by 36% when 2% methanol was added every 24 h. The influence of initial pH, temperature, methanol addition, precursors (menadione, isopentenol) addition, catalytic time and cetyltrimethyl-ammonium bromide (CTAB) addition were explored in the P. pastoris whole-cell catalytic synthesis process of MK-3 in shaking flask. Three significant factors were then studied by response surface method. The optimal catalytic conditions obtained were as follows: catalytic temperature 31.56 ℃, menadione 295.54 mg/L, catalytic time 15.87 h. Consistent with the response surface prediction results, the optimized yield of MK-3 reached 98.47 mg/L in shaking flask, 35% higher than that of the control group. On this basis, the production in a 30-L fermenter reached 189.67 mg/L when the cell catalyst of 220 g/L (dry weight) was used to catalyze the synthesis for 24 h. This method laid the foundation for the large-scale production of MK-3 by P. pastoris Gpn12.

Objective To investigate the effect ofdexmedetomidine (Dex) on endoplasmic reticulum stress (ERS) and apoptosis in brain injury after asphyxiating cardiac arrest and cardiopulmonary resuscitation (CA/CPR) in rats.Methods A total of 60 clean male Sprague-Dawley rars were randomly divided into sham-operated group,CA/CPR group and Dex precondition group (n=20).Rats in the control group did not receive CA/CPR;and rats in the CA/CPR group and Dex precondition group were performed cardiac arrest induced by asphyxia,and then,CPR was performed.Dex with dose of 4.0 microgram/kg (body weight) was intravenously injected into rats in the Dex precondition group prior to 5 min of asphyxia.The same volume of saline by intravenous injection was given to rats in control group and CA/CPR group.Brain tissues were collected after the experiment,and wet to dry weight (W/D) ratio was tested.The mRNA expressions of CCAAT-enhancer binding protein homologous protein (CHOP),activation of transcription factor 4 (A TF4) and X-4 box binding protein 1 (XBP1) in the hippocampus were detected by reverse transcription-polymerase chain reaction (RT-PCR).The protein expressions of CHOP,B-cell lymphoma-2 (Bcl-2),Bcl-2 associated X protein (Bax) and cysteinyl aspartate specific proteinase 3 (caspase-3) in the hippocampus were measured by Western boltting.Neuronal apoptosis was detected by terminal dexynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL).Morphological and ultrastructural changes of brains of rats were observed by light microscopy and electron microscopy.Results As compared with the control group,CA/CPR group and Dex precondition group had significantly increased W/D ratio of brain tissues and mRNA expressions of XBP1,A TF4 and CHOP in the hippocampus,significantly higher protein expressions of CHOP,Bax and caspase-3,and statistically lower Bcl-2 expression (P＜0.05).As compared with the CA/CPR group,Dex precondition group had significantly decreased W/D ratio of brain tissues and mRNA expressions of XBP1,A TF4 and CHOP in the hippocampus,significantly lower protein expressions of CHOP,Bax and caspase-3,and statistically higher Bcl-2 expression (P＜0.05).TUNEL indicated that the neuronal apoptosis rate in the control group (7.49％±4.33％) was significantly lower than that in the CA/CPR group and Dex precondition group (29.73％±6.27％ and 16.82％±5.75％,P＜0.05);significant difference was noted between CA/CPR group and Dex precondition group in the neuronal apoptosis rate (P＜0.05).Changes in the morphology and ultramicrostructure injuries of brain tissues were more significant in CA/CPR group,while the changes were obviously alleviated in Dex precondition group.Conclusion Dex can alleviate brain injury after asphyxiating CA/CPR in rats,whose mechanism may be related to ERS and inhibition on apoptosis of nerve cells.

Objective To increase the water solubility of curcumin constituents by three kinds of food grade surfactants, and to study the effect of surfactants on the quality standard and stability and anti-hepatic fibrosis in vivo of curcumin solution. Methods The high-speed dispersion method was used to prepare the solution of curcuminoids-surfactants. The qualitative analysis of curcuminoids was confirmed by LC-MS/MS. HPLC-DAD fingerprinting was established and quantitative analysis was conducted. And the effects on anti-liver fibrosis of different concentrations curcumin ratio solution were evaluated by rat model. Results The maximum water solubility of curcumin constituents was 1.387 mg/mL. LC-MS/MS analysis showed that the structure of curcuminoids were stable; The results of fingerprinting analysis demonstrated that the similarity of 18 batches of samples were higher than 99%. The contents of curcumin, demethoxycurcumin and bisdemethoxycurcumin in different batches of samples were stable with the RSD of 1.60%, 2.24%, and 3.74%. The rat model of hepatic fibrosis showed that the contents of AST and ALT in free curcumin suspension group were (64.1 ± 20.3) and (45.1 ± 13.9) U/L, respectively, while that solution of curcuminoids-surfactants group was (19.4 ± 8.7) and (11.8 ± 4.9) U/L. Conclusion The solubility of curcuminoids was increased 517 times compared with the free state (2.677 μg/mL) after solubilization with surfactants, and repeated tests indicated that the structure and contents of curcuminoids were stable; the solubilized curcuminoids exhibited better anti-liver fibrosis effects than the free.

Objective To evaluate the effect of dexmedetomidine on retinal ischemia-reperfusion (I/R) injury in mice.Metbods Forty-eight C57BL/6 male mice,aged 8 weeks,weighing 20-24 g,were divided into 3 groups (n =16 each) using a random number table:control group (C group),I/R group and dexmedetomidine group (D group).The model of retinal I/R injury was established by elevating intraocular pressure for 60 min using anterior chamber cannulation followed by 24 h of reperfusion.At 15 min before ischemia and 5 min before reperfusion,dexmedetomidine 25 μg/kg was intraperitoneally injected in group D,and the equal volume of normal saline was intraperitoneally injected in C and I/R groups.Eight mice were sacrificed at 24 h of reperfusion,and the retina was removed for microscopic examination of pathologic changes (with light microscope) after haematoxylin and eosin staining and for determination of cell apoptosis in retinal tissues (by TUNEL).Apoptosis index (AI) was calculated.Eight mice were sacrificed at 24 h of reperfusion,and the retina was removed for determination of the expression of Bcl-2,Bax,caspase-3 and CCAAT/enhancer-binding protein homologous protein (CHOP) in retinal tissues (by Western blot).Bcl-2/Bax ratio was calculated.Results Compared with group C,AI was significantly increased,the expression of Bax,caspase-3 and CHOP was up-regulated,the expression of Bcl-2 was down-regulated,Bcl-2/Bax ratio was decreased (P＜0.05),and pathologic changes were found in retinal tissues in group I/R.Compared with group I/R,AI was significantly decreased,the expression of Bax,caspase-3 and CHOP was down-regulated,the expression of Bcl-2 was up-regulated,Bcl-2/Bax ratio was increased (P＜0.05),and pathologic changes of retinal tissues were significantly attenuated in group D (P＜0.05).Conclusion Dexmedetomidine can reduce retinal I/R injury,and the mechanism mav be related to inhibiting cell apoptosis in mice.

OBJECTIVETo investigate the relationship between apoptosis-related proteins in gastric mucosa, p53 and Bax, and Helicobacter pylori (H. pylori) infection in children.

METHODSp53 and Bax expression in gastric mucosa were measured using immunohistochemical technique in 33 children with gastric mucosal lesions. Presence/absence of H. pylori infection was detected by the rapid urease and pathological tests.

RESULTSFifteen children (88%) showed positive expression of p53 in 17 children who were confirmed with H. pylori infection, compared with 9 (56%) in 16 H. pylori negative children. Thirteen children (76%) showed positive expression of Bax in the 17 children with H. pylori infection, compared with 6 (38%) in the 16 H. pylori negative children. The expression levels of p53 and Bax in the H. pylori positive group were significantly higher than those in the H. pylori negative group (p<0.05).

CONCLUSIONSH. pylori infection is associated with the over-expression of p53 and Bax proteins in gastric mucosa in children.

Child ; Female ; Gastric Mucosa ; chemistry ; Helicobacter Infections ; metabolism ; Helicobacter pylori ; Humans ; Immunohistochemistry ; Male ; Tumor Suppressor Protein p53 ; analysis ; bcl-2-Associated X Protein ; analysis