Objective To explore the possible mechanism of T cell immunoglobulin and mucin domain-containing protein-3(TIM-3)regulating M2 macrophage polarization through Galectin-3 in lung injury induced by Streptococcals pneumoniae pneumonia(SPP)in mice.Methods Thirty mice were divided into control group,SPP group and SPP+TIM-3 inhibition group,with 10 mice in each group.The macrophage cell line RAW 264.7 was cultured and divided into a control group,with the SPP group versus the SPP+TIM-3 inhibition group.Lung histopathology was detected by hematoxylin-eosin(H&E)staining.Serum and cell supernatant tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β)and IL-6 content were measured by enzyme-linked immunosorbent assay(ELISA).Western blotting the protein levels of TIM-3 and Galectin-3 were determined in lung tissues and macrophages.Cluster of differentiation(CD)206,CD86 mRNA levels in lung tissue and macrophages were determined by real time quantitative PCR(RT-qPCR).Results Compared with the control group,the SPP group had significantly abnormal lung tissue pathological structure,thickened alveolar septum,a large number of inflammatory cells infiltration,a large amount of exudate in alveolar space,and increased serum inflammatory factors TNF-α,IL-6 and IL-1β,the protein and mRNA expressions of CD86,the marker protein of M1 macrophages,and the protein and mRNA expressions of CD206,the marker protein of M2 macrophages,and the protein expressions of TIM-3 and Galectin-3 in lung tissues were significantly increased in the model group,and the differences were statistically significally(t=8.36～76.30,all P<0.05).Compared with the SPP group,the SPP+TIM-3 inhibition group had improved lung tissue pathological damage,reduced alveolar septum thickness,inflammatory cell infiltration,reduced exudate in alveolar space,decreased serum inflammatory factors TNF-α,IL-6 and IL-1β content,and decreased expression of CD86 protein and mRNA in lung tissue.The expression of CD206 protein and mRNA was increased,the expression of TIM-3 protein was decreased,and the expression of Galectin-3 protein was increased,and the differences were statistically significant(t=10.67～63.32,all P<0.05).Cell experiments were consistent with this result.Conclusion Inhibition of TIM-3 can improve lung injury in SPP mice by down-regulating the expression of Galectin-3 and promoting M2 macrophage polarization.

Objective To assess the efficacy of tandem mass spectrometry-specific indicators in diagnosing β-ketothiolase deficiency(BKD)and to elucidate the associated gene variations contributing to the corresponding pathogenic phenotype,thereby facilitating rapid and accurate diagnosis of BKD.Methods Data from tandem mass spectrometry(MS/MS)screening of dried blood spots collected from 16,071 children between January 2018 and December 2021,along with results from gas chromatography-mass spectrometry(GC-MS)analysis and high-throughput sequencing of positive cases,were analyzed retrospectively.The study aimed to evaluate the contribution of specific MS/MS indicators in the clinical diagnosis of BKD and to trace the genetic etiology of this disease.Results Among the 16 071 subjects screened by MS/MS,37 cases(2.30‰)exhibited elevated C5OH levels,41 cases(2.55‰)showed increased C5∶1 levels,and 2 cases were diagnosed with BKD based on GC-MS analysis.When diagnosing BKD using C5OH as a single indicator,the false positive rate was 0.22%,which is lower than that of C5∶1(0.24%).The positive predictive value for C5OH was 5.40%,higher than that of C5∶1(4.88%).Among the 16 071 pediatric patients,only 2 cases were diagnosed with BKD due to elevated C5OH combined with increased C5∶1 levels,resulting in a positive predictive value of 100%.Whole exome sequencing of these two BKD patients revealed that both carried acetyl-CoA acetyltransferase 1(ACAT1)gene double allele missense heterozygous mutations.The four previously unreported mutation sites were c.949G>C(p.Asp317His),c.1063G>A(p.Ala355Thr),c.146G>A(p.Arg49Lys),and c.700G>A(p.Glu234Lys).These findings provide novel insights into the genetic basis of BKD.Conclusions The MS/MS screening indicator C5OH demonstrates superior diagnostic efficacy compared to C5∶1 in diagnosing BKD,as evidenced by lower false positive rates and higher positive predictive values.When diagnosing BKD,the use of combined indicators significantly enhances the accuracy of biochemical diagnosis compared to single indicators.Exome sequencing revealed that all BKD patients carried previously unreported mutations in the ACAT1.

Objective To assess the efficacy of tandem mass spectrometry-specific indicators in diagnosing β-ketothiolase deficiency(BKD)and to elucidate the associated gene variations contributing to the corresponding pathogenic phenotype,thereby facilitating rapid and accurate diagnosis of BKD.Methods Data from tandem mass spectrometry(MS/MS)screening of dried blood spots collected from 16,071 children between January 2018 and December 2021,along with results from gas chromatography-mass spectrometry(GC-MS)analysis and high-throughput sequencing of positive cases,were analyzed retrospectively.The study aimed to evaluate the contribution of specific MS/MS indicators in the clinical diagnosis of BKD and to trace the genetic etiology of this disease.Results Among the 16 071 subjects screened by MS/MS,37 cases(2.30‰)exhibited elevated C5OH levels,41 cases(2.55‰)showed increased C5∶1 levels,and 2 cases were diagnosed with BKD based on GC-MS analysis.When diagnosing BKD using C5OH as a single indicator,the false positive rate was 0.22%,which is lower than that of C5∶1(0.24%).The positive predictive value for C5OH was 5.40%,higher than that of C5∶1(4.88%).Among the 16 071 pediatric patients,only 2 cases were diagnosed with BKD due to elevated C5OH combined with increased C5∶1 levels,resulting in a positive predictive value of 100%.Whole exome sequencing of these two BKD patients revealed that both carried acetyl-CoA acetyltransferase 1(ACAT1)gene double allele missense heterozygous mutations.The four previously unreported mutation sites were c.949G>C(p.Asp317His),c.1063G>A(p.Ala355Thr),c.146G>A(p.Arg49Lys),and c.700G>A(p.Glu234Lys).These findings provide novel insights into the genetic basis of BKD.Conclusions The MS/MS screening indicator C5OH demonstrates superior diagnostic efficacy compared to C5∶1 in diagnosing BKD,as evidenced by lower false positive rates and higher positive predictive values.When diagnosing BKD,the use of combined indicators significantly enhances the accuracy of biochemical diagnosis compared to single indicators.Exome sequencing revealed that all BKD patients carried previously unreported mutations in the ACAT1.

Objective To explore the possible mechanism of T cell immunoglobulin and mucin domain-containing protein-3(TIM-3)regulating M2 macrophage polarization through Galectin-3 in lung injury induced by Streptococcals pneumoniae pneumonia(SPP)in mice.Methods Thirty mice were divided into control group,SPP group and SPP+TIM-3 inhibition group,with 10 mice in each group.The macrophage cell line RAW 264.7 was cultured and divided into a control group,with the SPP group versus the SPP+TIM-3 inhibition group.Lung histopathology was detected by hematoxylin-eosin(H&E)staining.Serum and cell supernatant tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β)and IL-6 content were measured by enzyme-linked immunosorbent assay(ELISA).Western blotting the protein levels of TIM-3 and Galectin-3 were determined in lung tissues and macrophages.Cluster of differentiation(CD)206,CD86 mRNA levels in lung tissue and macrophages were determined by real time quantitative PCR(RT-qPCR).Results Compared with the control group,the SPP group had significantly abnormal lung tissue pathological structure,thickened alveolar septum,a large number of inflammatory cells infiltration,a large amount of exudate in alveolar space,and increased serum inflammatory factors TNF-α,IL-6 and IL-1β,the protein and mRNA expressions of CD86,the marker protein of M1 macrophages,and the protein and mRNA expressions of CD206,the marker protein of M2 macrophages,and the protein expressions of TIM-3 and Galectin-3 in lung tissues were significantly increased in the model group,and the differences were statistically significally(t=8.36～76.30,all P<0.05).Compared with the SPP group,the SPP+TIM-3 inhibition group had improved lung tissue pathological damage,reduced alveolar septum thickness,inflammatory cell infiltration,reduced exudate in alveolar space,decreased serum inflammatory factors TNF-α,IL-6 and IL-1β content,and decreased expression of CD86 protein and mRNA in lung tissue.The expression of CD206 protein and mRNA was increased,the expression of TIM-3 protein was decreased,and the expression of Galectin-3 protein was increased,and the differences were statistically significant(t=10.67～63.32,all P<0.05).Cell experiments were consistent with this result.Conclusion Inhibition of TIM-3 can improve lung injury in SPP mice by down-regulating the expression of Galectin-3 and promoting M2 macrophage polarization.

Objective: To explore the clinical efficacy of spine subtle adjusting manipulation for postpartum low back pain (PLBP). Methods: A total of 76 patients with PLBP were randomized into a control group and a treatment group, with 38 cases in each group. The control group was treated with core muscle strengthening exercises, and the treatment group was treated with spine subtle adjusting manipulation. After 3 weeks of treatment, the clinical efficacy was observed, and the visual analog scale (VAS) score and Oswestry disability index (ODI), and the changes of lumbar Cobb angle and pelvic rotation were compared between the two groups. Results: The total effective rate of the treatment group was 92.1％, and that of the control group was 78.9％. The difference between the two groups was statistically significant (P<0.05). After treatment, the VAS score and ODI in both groups decreased, and the intra-group differences were all statistically significant (P<0.05). There were no intra-group statistical differences in the lumbar Cobb angle or pelvic rotation in the two groups (P>0.05). After treatment, there were no statistical differences in the lumbar Cobb angle or pelvic rotation between the two groups (P>0.05); the VAS score and ODI in the treatment group were significantly lower than those in the control group (P<0.05). Conclusion: Spine subtle adjusting manipulation can effectively relieve the pain for patients with PLBP, and improve their daily activity function.

This article reviews the present situation, progress, challenges, and the future developments for the screening and diagnosis of inherited metabolic disease (IMD) in both domestic and abroad in clinical practice, aiming to improve the screening and diagnosis system of IMD in China and to provide a reference for clinicians to increase the diagnosis and cure rate of treatable IMD, and promote the development of this field.

Objective To compare the differences in metabolites between newborns with intrauterine growth restriction (IUGR) and appropriate for gestational age (AGA) in order to understand the changes in metabolites of newborns with IUGR and explore the possible metabolic mechanism of tissue and organ damages in patients with IUGR,with the ultimate goal of providing the basis for clinical intervention.Methods A total of 45 newborns with IUGR and 56 AGA newborns who were hospitalized in the Neonatal Intensive Care Unit of Bayi Children's Hospital,the General Hospital of the Chinese People's Liberation Army between July 2009 and June 2015 and who underwent metabolic disease screening were enrolled in this study.The differences in of 21 amino acids and 55 carnitines in peripheral blood,as well as the changes in the ratios of free carnitine and acylcarnitine to total carnitine,were compared.Results (1)According to the comparison of birth weights (＜ 3rd percentile,3rd-＜ 5th percentile,5th-＜ 10th percentile,and 10th-90th percentile),peripheral blood of the IUGR newborns with birth weight ＜ 3rd percentile contained lower concentrations of alanine (F =2.94,P =0.03),homocysteine (F =3.83,P =0.01),methionine (F =2.88,P =0.04),ornithine(F =3.32,P =0.02),serine (F =3.09,P =0.03) and tyrosine (F =4.76,P =0.00) than those of the AGA newborns.In the peripheral blood of the IUGR newborns with birth weight of 3rd-＜ 5th percentile,the diversity of alanine concentrations showed compensatory increase,and their alanine concentrations were higher than those of the AGA newborns.(2) Metabolites also had significant differences in different gestational age groups:the concentrations of alanine (t =2.423,P =0.026),proline (t =2.470,P =0.023),and 14-carbon acylcarnitine (t =-2.870,P =0.010) in premature was higher than those in full-term newborns,but the concentration of 26-carbon acylcarnitine (t =-2.189,P =0.041) was lower than full-term ones;the concentrations of alanine (t =2.354,P =0.022),glutamine (t =2.520,P =0.015),pipecolic acid (t =2.017,P =0.049),proline (t =2.204,P =0.032) in premature AGA newborns were higher than those in full-term ones,but the concentrations of homocysteine (t =-2.624,P =0.011),seven carbon acylcarnitine(t =-2.403,P =0.020),and ten carbon acylcarnitine (t =-5.739,P =0.000) were lower than those of full-term AGA newborns;the concentrations of homocysteine (t =-2.421,P =0.020),decanogl carnitine(t =-2.181,P =0.035),methyl propylene acyl carnitine (t =-2.373,P =0.022),pentyl acyl carnitine (t =-2.165,P =0.036),decyl acyl carnitine (t =-4.148,P =0.000),hydroxyl acetyl carnitine (t =-2.097,P =0.042),hydroxyl cetyl acylcarnitine (t =-2.446,P =0.019) in premature IUGR were higher than those in fullterm IUGR newborns;but the concentrations of arginine (t =2.167,P =0.036),glutamic acid (t =2.469,P =0.018),histidine (t =2.718,P =0.009),leucine/isoleucine (t =3.938,P =0.000),ornithine (t =4.264,P =0.000),serine (t =2.647,P =0.011),threonine (t =2.311,P =0.026),tryptophan (t =4.040,P =0.000),valine (t =2.700,P =0.01),7-carbon acylcarnitine (t =-2.44 1,P =0.019),18-carbon diene carnitine (t =2.449,P =0.018),capric acylcarnitine(t =-4.148,P =0.000) and hydroxyl acetyl carnitine (t =-2.097,P =0.042) were lower than those in full-term IUGR newborns.(3) For AGA newborns,metabolites had no differences between male and female (P ＞ 0.05);however,for newborns with IUGR,metabolites significantly differed between male and female,and the concentrations of aspartic acid(t=2.521,P =0.016),glutamate(t =-2.175,P =0.035) in male IUGR were lower than those in female newborns with IUGR,but the concentration of 26-carbon carnitine (t =2.231,P =0.031) was higher than that in female group.(4) Birth weight had no significant effect on free carnitine concentration or on the ratios of free carnitine and acylcarnitine to total carnitine(all P ＞ 0.05).Conclusions IUGR infants exhibit significant abnormalities in amino acid and acylcarnitine metabolism,especially those with birth weight ＜ 3rd percentile.With the increase of birth weight,amino acids and acylcarnitines showed compensatory increases or decrease,and when birth weight reached the 10th percentile,the newborns with IUGR were close to the AGA newborns.

Objective To study the clinical distribution of Listeria monocytogenes infection and the changes in drug resistance of Listeria monocytogenes isolated from inpatients during recent 3 years,and to increase the awareness of the situation and provide data for clinical antibiotics application.Methods The clinical distribution of 22 cases of neonatal Listeria infection and drug resistance changes of Listeria were retrospectively analyzed in Bayi Children's Hospital from Jan.2011 to Dec.2013.Results Neonates began to be attacked by Listeria monocytogenes of 0.5 hours to 5 days (an average of 17.45 hours) after birth.The average birth weight was (2 331.82 ± 677.64) g.There were 7 full term cases and 15 premature infants,13 cases with low birth wcight.The average hospitalization was (21.91 ± 17.64)days.The cure rate was 45.45％ (10/22 cases).All the mothers of 15 cases had fever in the third trimester of pregnancy and the temperature was 37.5-39.5 ℃.Infection rate with Listeria monocytogenes in neonatal was 0.03％ (2/7 137 cases),0.11％ (8/7 281 cases) and 0.19％ (12/6 394 cases) in 3 years,respectively.From 2011 to 2013,the sensitive rate of antimicrobial drugs with Listeria monocytogencs to commonly used antimicrobial was 82.72％,75.40％ and 50.66％,and the rate of drug resistance was 17.28％,17.50％ and 11.01％,respectively.During 3 years,the rates of drug resistance had no significant difference (x2 =3.65,P ＞ 0.05),and the sensitive rates had a trend of declination year by year(x2 =36.87,P ＜ 0.01).The sensitive rates and the drugs resistant rates of penicillin were 33.93％ (19/56 cases)and 51.79％ (29/56 cases),respectively.In 3 years,the drugs resistant rates of penicillin was 100.00％,40.00％,and 46.43％,and the sensitive rate was 0,60.00％,25.00％,respectively.There was a high sensitivity of Listeria monocytogenes to ampicillin,aminoglycoside,sugar peptide,tetracycline,macrolides,lincosamides,quinolone,sulfa and other classes (such as rifampicin).It showed the different drug resistance rates with 33.33％-100.00％ to oxacillin,penicillin G and nitrofurans.Conclusions These children has the characteristics of early-onset infection.The pregnant women and newborns are susceptible to high-risk groups.Infection rates with Listeria of neonatal and Listeria monocytogenes isolated from inpatients showed a trend of increase year by year.The cases were very sensitive to commonly used antimicrobial for killing Listeria monocytogenes.There was a trend of the declination for drug resistance to penicillin,but it was still at a higher level.The drugs resistance rate to oxacillin,penicillin G and nitrofurans were high.

Objective To learn the incidence of the inherited metabolic diseases in Beijing. Methods Urine samples were analyzed by gas chromatography-mass spectrometry(GC-MS)for inherited metabolic diseases in high risky infants in Beijing . Results Urine samples from 411 high risky infants were analyzed by gas chromatography-mass spectrometry. 269 cases (65.5%) were detected to have metabolic abnormalities, including 19 cases (4.6%) diagnosed of inherited metabolic diseases in which there were 15 cases of methylmalonic academia and 1 case each of propionic academia, hyperphenylalaninemia, urea cycle abnormality and pyroglutamic aciduria. There were 22 suspected cases (5.4%) of inherited metabolic diseases including 13 cases of lactic acidosis, 5 cases of primary glycerol aciduria, 4 cases of fatty acid metabolic disorders including 1 case each of Citrin defects, tyrosinemia, galactosemia 3-methylcrotonoyl coenzyme A carboxylase deifciency and maple syrup urine disease. There were also 228 cases (55.5%) of metabolic abnormalities, such as increasing urine levels of lactic acid, sucrose,lactose, galactose, N-acetyl tyrosine, succinic acid, dicarboxylic acid and abnormal serine/threonine ratio. Conclusions Methylmalonic academia might be the most common inherited metabolic diseases in high risky infants in Beijing. For infants with clinical manifestations but unclear etiology, GC-MS should be performed. MS-MS and gene analysis could be combined if necessary.

mRNA display provides a new powerful tool for in vitro selecting of peptides and proteins.In the selecting process,peptides are covalently attached to their own mRNA to form mRNA-protein fusions.These mRNA-protein fusions enable in vitro selection of peptide and protein libraries of more than 1013 different sequences.The experiment conditions and protocols have been optimized in recent years.The application of mRNA display technology is mainly in the discovery of ligands for many kinds of target molecules and the analysis and mechanism elucidation on interaction between proteins.With its great potential,there will be a wide application foreground in the application of mRNA display.