Objective To investigate the mechanism by which Shenqi Xiaozheng Decoction-medicated serum regulates c-Myc/SLC1A5/GLS1 signaling axis to inhibit glutamine(Gln)metabolism and activate immunogenic cell death(ICD)in non-small cell lung cancer(NSCLC)cells.Methods A549 cells were divided into control group,model group,Shenqi Xiaozheng Decoction-medicated serum low-,medium-and high-dosage groups and positive control group.A Gln-dependent growth model was established,and cells were treated with different concentrations of Shenqi Xiaozheng Decoction-medicated serum or the SLC1A5 inhibitor V-9302.Cell proliferation was assessed by CCK-8,EdU and colony formation assays;Cell invasion and migration were evaluated using Transwell and wound-healing assays;intracellular Gln,glutathione(GSH),and α-ketoglutarate(α-KG)contents were determined by colorimetric assay;reactive oxygen species(ROS)contents were measured with fluorescent probes;Western blot was used to detect the protein expressions of E-cadherin,N-cadherin,c-Myc,SLC1A5 and GLS1;c-Myc/SLC1A5 colocalization and high mobility group box 1(HMGB1)expression were assessed by dual immunofluorescence staining;flow cytometry was used to evaluate calreticulin(CRT)exposure on the cell surface,and ATP and HMGB1 contents in cell supernatants were quantified by ELISA.Results Compared with the control group,the model group showed significantly increased A549 cell viability,EdU-positive rate and migration rate(P<0.05),as well as higher colony counts and invasion cell numbers(P<0.05);cellular Gln,GSH and α-KG contents were significantly elevated(P<0.05,P<0.01),while ROS content were not significantly different(P>0.05),E-cadherin protein expression significantly decreased,whereas the protein expressions of N-cadherin,c-Myc,SLC1A5 and GLS1 significantly increased(P<0.05,P<0.01).c-Myc and SLC1A5 colocalization was enhanced,HMGB1 expression was significantly increased(P<0.01),CRT exposure significantly increased(P<0.01),and ATP and HMGB1 contents in cell supernatant were significantly elevated(P<0.05,P<0.01).Compared with the model group,Shenqi Xiaozheng Decoction-medicated serum at different concentrations significantly inhibited Gln-stimulated A549 cell proliferation,migration and invasion in a dosage-dependent manner.Mechanistic studies indicated that Shenqi Xiaozheng Decoction could reduce Gln uptake and synthesis of its metabolic products GSH and α-KG,induce ROS accumulation,up-regulate protein expression of E-cadherin,down-regulate the protein expressions of N-cadherin,c-Myc,SLC1A5 and GLS1(P<0.05,P<0.01),and enhance CRT exposure,ATP release and HMGB1 secretion(P<0.01).Conclusion Shenqi Xiaozheng Decoction may exert a synergistic"metabolism-immunity"antitumor effect by inhibiting c-Myc/SLC1A5/GLS1 axis-mediated Gln uptake,inducing ROS accumulation,and activating ICD signaling.

Objective To evaluate the efficacy and safety of Qixian Tongluo Formula in the treatment of post-cerebral infarction paralysis with kidney deficiency and blood stasis syndrome,and to preliminarily explore the molecular mechanism of Qixian Tongluo Formula in improving impaired motor function from the perspective of cross-kingdom regulation of Chinese medicine microRNA(miRNA).Methods A pragmatic randomized controlled trial was conducted with 102 patients in the recovery period of post-cerebral infarction paralysis with kidney deficiency and blood stasis syndrome in our hospital.Patients were randomly divided into trial group and control group,with 51 cases in each group.The control group received standard Western medicine standard treatment,while the trial group received Qixian Tongluo Formula in addition to the standard treatment,with one dose per day,boiled in water,and taken warm after breakfast and dinner for a course of 2 months.The disability rate was used as the main efficacy indicator,and the incidence of adverse reactions was used as a safety indicator.miRNA from patient serum and Qixian Tongluo decoction were extracted respectively,and high-throughput sequencing was performed.The two sequences were compared to screen out the cross-kingdom gene transfer of Chinese medicine miRNA.Finally,its target genes of miRNA were predicted,and GO function and KEGG pathway enrichment analysis were carried out.Results A total of 67 patients completed the clinical trial,including 36 cases in the trial group and 31 cases in the control group;The disability rate in the trial group(13.9%)was lower than that in the control group(35.5%)(P<0.05);The incidence of adverse reactions was similar between the trial group(7.69%)and the control group(6.06%)(P>0.05);A total of 9530 Qixian Tongluo decoction miRNA sequences were screened,with 150 potentially involved in cross-kingdom gene transfer,including families such as miR-15 and miR-17;According to the target gene prediction of the top 10 miRNAs in cross-kingdom gene transfer of Chinese medicine,345 overlapping target genes were obtained;GO functional enrichment analysis revealed 16 biological processes,7 cellular components,and 2 molecular functions among the top 25 enriched functions,while KEGG pathway analysis mainly focused on the transforming growth factor-βsignaling pathway,neurotrophin signaling pathway,which are closely related to neural repair and functional recovery processes such as glial scar formation and synaptic plasticity after cerebral ischemia.Conclusion Qixian Tongluo Formula can significantly improve the functional independence level of patients with kidney deficiency and blood stasis syndrome in the recovery period of paralysis after cerebral infarction,offering a safe and effective treatment option for these patients;There were a large number of miRNAs in Qixian Tongluo decoction,some of which could cross-kingdom transferred into the human blood circulation,and promote the recovery of motor function in patients with cerebral infarction through multi-target,multi link and multi pathway gene network regulation.This study provides a new idea for subsequent clinical and basic research.

Objective To investigate the mechanism by which Shenqi Xiaozheng Decoction-medicated serum regulates c-Myc/SLC1A5/GLS1 signaling axis to inhibit glutamine(Gln)metabolism and activate immunogenic cell death(ICD)in non-small cell lung cancer(NSCLC)cells.Methods A549 cells were divided into control group,model group,Shenqi Xiaozheng Decoction-medicated serum low-,medium-and high-dosage groups and positive control group.A Gln-dependent growth model was established,and cells were treated with different concentrations of Shenqi Xiaozheng Decoction-medicated serum or the SLC1A5 inhibitor V-9302.Cell proliferation was assessed by CCK-8,EdU and colony formation assays;Cell invasion and migration were evaluated using Transwell and wound-healing assays;intracellular Gln,glutathione(GSH),and α-ketoglutarate(α-KG)contents were determined by colorimetric assay;reactive oxygen species(ROS)contents were measured with fluorescent probes;Western blot was used to detect the protein expressions of E-cadherin,N-cadherin,c-Myc,SLC1A5 and GLS1;c-Myc/SLC1A5 colocalization and high mobility group box 1(HMGB1)expression were assessed by dual immunofluorescence staining;flow cytometry was used to evaluate calreticulin(CRT)exposure on the cell surface,and ATP and HMGB1 contents in cell supernatants were quantified by ELISA.Results Compared with the control group,the model group showed significantly increased A549 cell viability,EdU-positive rate and migration rate(P<0.05),as well as higher colony counts and invasion cell numbers(P<0.05);cellular Gln,GSH and α-KG contents were significantly elevated(P<0.05,P<0.01),while ROS content were not significantly different(P>0.05),E-cadherin protein expression significantly decreased,whereas the protein expressions of N-cadherin,c-Myc,SLC1A5 and GLS1 significantly increased(P<0.05,P<0.01).c-Myc and SLC1A5 colocalization was enhanced,HMGB1 expression was significantly increased(P<0.01),CRT exposure significantly increased(P<0.01),and ATP and HMGB1 contents in cell supernatant were significantly elevated(P<0.05,P<0.01).Compared with the model group,Shenqi Xiaozheng Decoction-medicated serum at different concentrations significantly inhibited Gln-stimulated A549 cell proliferation,migration and invasion in a dosage-dependent manner.Mechanistic studies indicated that Shenqi Xiaozheng Decoction could reduce Gln uptake and synthesis of its metabolic products GSH and α-KG,induce ROS accumulation,up-regulate protein expression of E-cadherin,down-regulate the protein expressions of N-cadherin,c-Myc,SLC1A5 and GLS1(P<0.05,P<0.01),and enhance CRT exposure,ATP release and HMGB1 secretion(P<0.01).Conclusion Shenqi Xiaozheng Decoction may exert a synergistic"metabolism-immunity"antitumor effect by inhibiting c-Myc/SLC1A5/GLS1 axis-mediated Gln uptake,inducing ROS accumulation,and activating ICD signaling.

Objective To evaluate the efficacy and safety of Qixian Tongluo Formula in the treatment of post-cerebral infarction paralysis with kidney deficiency and blood stasis syndrome,and to preliminarily explore the molecular mechanism of Qixian Tongluo Formula in improving impaired motor function from the perspective of cross-kingdom regulation of Chinese medicine microRNA(miRNA).Methods A pragmatic randomized controlled trial was conducted with 102 patients in the recovery period of post-cerebral infarction paralysis with kidney deficiency and blood stasis syndrome in our hospital.Patients were randomly divided into trial group and control group,with 51 cases in each group.The control group received standard Western medicine standard treatment,while the trial group received Qixian Tongluo Formula in addition to the standard treatment,with one dose per day,boiled in water,and taken warm after breakfast and dinner for a course of 2 months.The disability rate was used as the main efficacy indicator,and the incidence of adverse reactions was used as a safety indicator.miRNA from patient serum and Qixian Tongluo decoction were extracted respectively,and high-throughput sequencing was performed.The two sequences were compared to screen out the cross-kingdom gene transfer of Chinese medicine miRNA.Finally,its target genes of miRNA were predicted,and GO function and KEGG pathway enrichment analysis were carried out.Results A total of 67 patients completed the clinical trial,including 36 cases in the trial group and 31 cases in the control group;The disability rate in the trial group(13.9%)was lower than that in the control group(35.5%)(P<0.05);The incidence of adverse reactions was similar between the trial group(7.69%)and the control group(6.06%)(P>0.05);A total of 9530 Qixian Tongluo decoction miRNA sequences were screened,with 150 potentially involved in cross-kingdom gene transfer,including families such as miR-15 and miR-17;According to the target gene prediction of the top 10 miRNAs in cross-kingdom gene transfer of Chinese medicine,345 overlapping target genes were obtained;GO functional enrichment analysis revealed 16 biological processes,7 cellular components,and 2 molecular functions among the top 25 enriched functions,while KEGG pathway analysis mainly focused on the transforming growth factor-βsignaling pathway,neurotrophin signaling pathway,which are closely related to neural repair and functional recovery processes such as glial scar formation and synaptic plasticity after cerebral ischemia.Conclusion Qixian Tongluo Formula can significantly improve the functional independence level of patients with kidney deficiency and blood stasis syndrome in the recovery period of paralysis after cerebral infarction,offering a safe and effective treatment option for these patients;There were a large number of miRNAs in Qixian Tongluo decoction,some of which could cross-kingdom transferred into the human blood circulation,and promote the recovery of motor function in patients with cerebral infarction through multi-target,multi link and multi pathway gene network regulation.This study provides a new idea for subsequent clinical and basic research.

Objective To observe the effect of Qixian Tongluo Formula on contralateral corticospinal tract(CST)remodeling and motor functional recovery in rats with cerebral infarction,and to explore its potential molecular mechanism from the perspective of regulating factors related to never remodeling.Methods The rat middle cerebral artery occlusion(MCAO)model was established by silk thread ligation.Fifty model rats were randomly divided into model group,citicoline group(0.054 g·kg-1),Qixian Tongluo Formula low-,medium-and high-dose(7.83,15.66,31.32 g·kg-1)groups,and sham operation group,with 10 rats in each group.The intervention administration was started on the 3rd day after operation once a day for 26 consecutive days.On the 3rd,14th and 28th day after operation,the gross motor function was evaluated by Longa score,and the fine motor function was evaluated by beam-walking test(BWT)score.The contralateral motor cortex was injected with the nerve tracer biotin dextran amine(BDA)on the 14 th day after operation to anterogradely trace the CST.On the 28th day after operation,the expression of axonal growth associated protein-43(GAP-43)and BDA positive fibers in the contralateral motor cortex and cervical spinal cord were detected by immunohistochemistry.The co-localization areas of BDA positive fibers and presynaptic marker protein vesicular glutamate transporter 1(VGLUT1)in the cervical spinal cord gray matter were detected by immunofluorescence.The expressions of brain-derived neurotrophic factor(BDNF),glial cell-derived neurotrophic factor(GDNF),nerve growth factor(NGF)and nerve remodeling-associated inhibitory factor[Nogo-A,oligodendrocyte myelin glycoprotein(OMgp)and myelin-associated glycoprotein(MAG)]in the contralateral motor cortex were detected by Western Blot.Pearson correlation analysis was used to analyze the correlation between Longa score or BWT score and BDA/VGLUT1 co-localization area,respectively.Results Compared with the sham operation group,rats in the model group had obvious symptoms of motor function deficits,and the Longa scores were significantly increased(P＜0.01)and the BWT scores were significantly decreased(P＜0.05,P＜0.01)at each time point.The expression of GAP-43 in the contralateral motor cortex and cervical spinal cord was up-regulated(P＜0.05),the number of edge-crossing fibers from the posterior funiculus in cervical cord was increased(P＜0.05),the co-localization area of BDA/VGLUT1 in the gray matter of the cervical spinal cord was increased(P＜0.05),the expressions of BDNF,GDNF and NGF in the contralateral motor cortex were up-regulated(P＜0.05),while the expressions of Nogo-A,OMgp and MAG were down-regulated(P＜0.05).Compared with the model group,the Longa scores in each administration group on the 14th and 28th day after MCAO operation were significantly decreased(P＜0.01),the BWT scores were significantly increased(P＜0.01),the expression of GAP-43 in the contralateral motor cortex and cervical spinal cord was significantly up-regulated(P＜0.01).The number of edge-crossing fibers from the posterior funiculus in cervical cord was significantly increased(P＜0.01),the co-localization area of BDA/VGLUT1 in the gray matter of the cervical spinal cord was significantly increased(P＜0.01).The expressions of BDNF,GDNF and NGF in the contralateral motor cortex were significantly up-regulated(P＜0.01,P＜0.05),while the expressions of Nogo-A,OMgp and MAG were significantly down-regulated(P＜0.05),and the most significant effect was observed in the high dose group.The Longa score was negatively correlated with the co-localization area of BDA/VGLUT1(r=-0.89,P＜0.01),and the BWT score was positively correlated with the co-localization area of BDA/VGLUT1(r=0.84,P＜0.01).Conclusion Qixian Tongluo Formula can improve motor function through promoting contralateral CST remodeling in MCAO rats after cerebral infarction,and the molecular mechanism may be related to the regulation of the expression of nerve remodeling-associated factor in the contralateral motor cortex.

Objective:To investigate the effect of lanatoside C on autophagy in hepatocellular carcinoma Huh-7 cells,and the possible mechanism of action.Methods:After Huh-7 cells were treated with gradient concentration of lanatoside C for 24 h,the IC50 value of lanatoside C on Huh-7 cells was detected by CCK-8 assay,and the role of intervention of gradient concentration of lanatoside C in the proliferation and clone formation of Huh-7 cells was analyzed by CCK-8 assay and clone formation assay.Western blotting was used to detect the expression levels of autophagy markers in Huh-7 cells,such as microtubule-associated protein light chain 3(LC3)-II,LC3-Ⅰ,p62,Beclin1,vacuolar protein sorting 34(VPS30),VPS15 and autophagy-related protein 3(ATG3)in Huh-7 cells,and the changes of LC3-II/LC3-Ⅰ ratio were analyzed.The effects of lanatoside C on the formation of autophagic vesicles in Huh-7 cells were observed by laser confocal microscopy,and the effects of lanatoside C on the formation of autophagic vesicles in Huh-7 cells were confirmed by artificial intelligence image analysis.Finally,the effects of lanatoside C on the interaction between Beclin1,VPS34 and VPS15 on the formation of autophagic vesicles in Huh-7 cells were detected by co-immunoprecipitation.Results:The IC50 value of lanatoside C on Huh-7 cells at 24 h was(0.49±0.07)μg/mL.The results of the CCK-8 assay and clone formation assay suggested that lanatoside C had a significant inhibitory effect on the growth of Huh-7 cells in a concentration-dependent manner(both P<0.01).The results of Western blotting assay showed that the LC3-II/LC3-Ⅰ ratio was significantly increased with the increase in the concentration of lanatoside C(P<0.01),while the expression level of Beclin1 protein was up-regulated(P<0.05),and the expression level of p62 protein was significantly down-regulated(P<0.01).The results of laser confocal microscopy showed that lanatoside C could significantly induce autophagic vesicle formation in Huh-7 cells(P<0.01).The results of artificial intelligence image analysis confirmed this effect of lanatoside C(all P<0.01).The results of co-immunoprecipitation confirmed that lanatoside C was able to promote the binding of Beclin1 to VPS34 and VPS15 with each other(both P<0.01).Conclusion:Lanatoside C significantly inhibits the proliferation of Huh-7 cells and and induces Huh-7 cells autophagy,the mechanism of this effect may be related to up-regulating Beclin1 expression and promoting the combination of Beclin1 protein with VPS34 and VPS15 proteins.

Objective:To investigate the effect of lanatoside C on autophagy in hepatocellular carcinoma Huh-7 cells,and the possible mechanism of action.Methods:After Huh-7 cells were treated with gradient concentration of lanatoside C for 24 h,the IC50 value of lanatoside C on Huh-7 cells was detected by CCK-8 assay,and the role of intervention of gradient concentration of lanatoside C in the proliferation and clone formation of Huh-7 cells was analyzed by CCK-8 assay and clone formation assay.Western blotting was used to detect the expression levels of autophagy markers in Huh-7 cells,such as microtubule-associated protein light chain 3(LC3)-II,LC3-Ⅰ,p62,Beclin1,vacuolar protein sorting 34(VPS30),VPS15 and autophagy-related protein 3(ATG3)in Huh-7 cells,and the changes of LC3-II/LC3-Ⅰ ratio were analyzed.The effects of lanatoside C on the formation of autophagic vesicles in Huh-7 cells were observed by laser confocal microscopy,and the effects of lanatoside C on the formation of autophagic vesicles in Huh-7 cells were confirmed by artificial intelligence image analysis.Finally,the effects of lanatoside C on the interaction between Beclin1,VPS34 and VPS15 on the formation of autophagic vesicles in Huh-7 cells were detected by co-immunoprecipitation.Results:The IC50 value of lanatoside C on Huh-7 cells at 24 h was(0.49±0.07)μg/mL.The results of the CCK-8 assay and clone formation assay suggested that lanatoside C had a significant inhibitory effect on the growth of Huh-7 cells in a concentration-dependent manner(both P<0.01).The results of Western blotting assay showed that the LC3-II/LC3-Ⅰ ratio was significantly increased with the increase in the concentration of lanatoside C(P<0.01),while the expression level of Beclin1 protein was up-regulated(P<0.05),and the expression level of p62 protein was significantly down-regulated(P<0.01).The results of laser confocal microscopy showed that lanatoside C could significantly induce autophagic vesicle formation in Huh-7 cells(P<0.01).The results of artificial intelligence image analysis confirmed this effect of lanatoside C(all P<0.01).The results of co-immunoprecipitation confirmed that lanatoside C was able to promote the binding of Beclin1 to VPS34 and VPS15 with each other(both P<0.01).Conclusion:Lanatoside C significantly inhibits the proliferation of Huh-7 cells and and induces Huh-7 cells autophagy,the mechanism of this effect may be related to up-regulating Beclin1 expression and promoting the combination of Beclin1 protein with VPS34 and VPS15 proteins.

Objective:To investigate the application value of P-loop digestive tract recons-truction in pancreaticoduodenectomy (PD).Methods:The retrospective and descriptive study was conducted. The clinicopathological data of 21 ampullary disease patients undergoing PD in the Liuzhou People′s Hospital Affiliated to Guangxi Medical University from April to December 2020 were collected. There were 13 males and 8 females, aged from 35 to 76 years, with a median age of 60 years. All the 21 patients underwent PD and digestive tract reconstruction using P-loop method based on the Child reconstruction. Observation indicators: (1) surgical situations; (2) postoperative situations; (3) follow-up. Follow-up was conducted using outpatient examination or telephone interview to detect survival and discomfort symptoms of patients up to December 2020. Measurement data with normal distribution were represented as Mean± SD, and measurement data with skewed distribution were represented as M(range). Count data were described as absolute numbers or persentages. Results:(1) Surgical situations: all the 21 patients underwent PD successfully. The operation time, time of P-loop anastomosis and volume of intraoperative blood loss of 21 patients were (317±74)minutes, (14±3)minutes and 375 mL(range, 100-800 mL), respectively. Of the 21 patients, 17 cases had pancreatic texture as soft, 4 cases had pancreatic texture as hard, 3 cases had diameter of pancreas ≤3 mm, 18 cases had diameter of pancreas >3 mm, 14 cases were placed pancreatic duct stent, 7 cases were not placed pancreatic duct stent. (2) Postoperative situations: 2 of the 21 patients had grade A pancreatic fistula, and none of patient had grade B or grade C pancreatic fistula. One case had hepaticojejunal anastomotic fistula, 2 cases without pancreatic fistula had delayed gastric emptying and none of patient had abdominal infection or bleeding. The duration of postoperative hospital stay of 21 patients was (16±5)days, and none of patient died during postoperative 30 days. Results of postoperative histopathological examination showed there were 10 cases with duodenal papillary carcinoma, 4 cases with lower bile duct carcinoma, 3 cases with pancreatic head ductal adenocarcinoma, 1 case with duodenum stromal tumors, 1 case with gastric antrum carcinoma, 1 case with mass in the head of the pancreas of IgG4 and 1 case with choledochal cyst of type 3. (3) Follow-up: all 21 patients were followed up for 1.0 to 7.0 months, with a median follow-up time of 4.3 months. None of patient died. There was no abdominal pain, distension or dyspepsia during follow-up. One case was diagnosed as tumor liver metastasis at postoperative 5 months.Conclusion:P-loop digestive tract reconstruction in PD is safe and effective, with good short-term effect.

A simple and sensitive assay for rapid detection of human coronavirus NL63 (HCoV-NL63) was developed by colorimetic reverse transcription loop-mediated isothermal amplification (RT-LAMP). The method employed six specially designed primers that recognized eight distinct regions of the HCoV-NL63 nucleocapsid protein gene for amplification of target sequences under isothermal conditions at 63 degrees C for 1 h Amplification of RT-LAMP was monitored by addition of calcein before amplification. A positive reaction was confirmed by change from light-brown to yellow-green under visual detection. Specificity of the RT-LAMP assay was validated by cross-reaction with different human coronaviruses, norovirus, influenza A virus, and influenza B virus. Sensitivity was evaluated by serial dilution of HCoV-NL63 RNA from 1.6 x 10(9) to 1.6 x 10(1) per reaction. The RT-LAMP assay could achieve 1,600 RNA copies per reaction with high specificity. Hence, our colorimetric RT-LAMP assay could be used for rapid detection of human coronavirus NL63.
Colorimetry ; methods ; Coronavirus Infections ; diagnosis ; virology ; Coronavirus NL63, Human ; genetics ; isolation & purification ; DNA Primers ; genetics ; Humans ; Nucleic Acid Amplification Techniques ; methods ; Reverse Transcription ; Sensitivity and Specificity

Objective To study the influence of protein intake in learning and memory capabilities and mammalian target of rapamycin (mTOR) expression in premature rats.Methods Premature rars were randomly divided into three groups after weaning:preterm standard protein group,preterm low protein group and preterm high protein group.After weaning,three groups were respectively fed with standard protein diets (18％ protein),low protein diets (8％ protein) and high protein diets (30％ protein) until the end of experiment.When rats were at the 6th and 8th weeks,they were performed Morris water maze test to assess the learning and memory abilities,and irnmunohistochemistry and Westem blotting were used to observe the mTOR distribution and mTOR protein expression in the hippocampus.Results At the age of 6 weeks,the escape latencies of the premature rats did not change significantly on the 3rd and 4th day of the experiment; the frequency of crossing the platform and the mTOR-positive cells in hippocampal CA1 and CA3 regions of each group showed no significant differences (P＞0.05).At the age of 8 weeks,the escape latencies in finding the platform were shortened with increased training times for premature rats; the frequency of crossing the platform of high-protein premature rats was significantly higher than that in the other two groups (P＜0.05); and the mTOR expression in the hippocampal CA1 and CA3 regions of high-protein premature rats was also significantly increased as compared with the other two groups (P＜0.05).High-protein premature rats at the age of 8 weeks had significantly higher frequency of crossing the platform and increased mTOR-positive cells in the hippocampal CA1 and CA3 regions than those at the age of 6 weeks (P＜0.05).Western blotting also indicated that the hippocampal mTOR protein expression was not significantly different in each group at 6 week-old; the mTOR expression of high-protein premature rats was significantly higher than that in the other two groups at the age of 8 weeks.Conclusions In the early growth and development process of the brain,mTOR activity might be inhibited in the premature birth rats,which can lead to impaired learning and memory abilities.After a long time of high protein nutritional intake,impaired learning and cognitive abilities of premature rats can get recovery,which may be associated with up-regulation ofmTOR expression.