Objective To explore the effects of L-carnitine combined with citicoline on the therapeutic effect of neonatal hypoxic-ischemic encephalopathy(NHIE)and serum nitric oxide(NO)and endothelin-1(ET-1).Methods A total of 95 children with HIE admitted to the hospital from April 2020 to January 2023 were selected and were divided into observation group(47 cases)and control group(48 cases)by random number table method.The control group was given citicoline,and the observation group was given citicoline combined with L-carnitine.The therapeutic effect was evaluated on 14 days after treatment,and the recovery time of original reflex,muscle tone and consciousness was calculated.The levels of oxidative stress indexes[malondialdehyde(MDA),superoxide dismutase(SOD)]and vascular endothelial function indexes(NO,ET-1)were detected before treatment and 14 days after treatment.Neonatal neurobehavioral score(NBNA)was used to evaluate the neurological function of the children,and the safety of the treatment regimen was ob-served.Pearson correlation analysis was used to analyze the correlation between NBNA and vascular endothe-lial function indexes.Results The total effective rate in the observation group was 91.49％,which was higher than 72.92％in the control group(P＜0.05).The original reflex recovery time,muscle tone recovery time and consciousness recovery time of NHIE children in the observation group were shorter than those in the control group(P＜0.05).There was no significant difference in NBNA scores between the two groups before treatment(t=1.225,P=0.224).After treatment,the NBNA score in the observation group was higher than that in the control group(t=6.223,P＜0.001).After treatment,MDA level decreased and SOD level in-creased in two groups(P＜0.05).After treatment,the level of MDA in the observation group was lower than that in the control group,while the level of SOD was higher than that in the control group(P＜0.05).After treatment,the levels of NO and ET-1 were decreased in both groups(P＜0.05).The levels of NO and ET-1 in the observation group were lower than those in the control group after treatment(P＜0.05).The adverse drug reaction rates in observation group and control group were 17.02％and 8.33％,respectively,and there was no significant difference between two groups(P＞0.05).Pearson correlation analysis showed that serum NO,ET-1 and NBNA score in NHIE children were negatively correlated(r=-0.546,-0.608,P＜0.05).Conclusion L-carnitine combined with citicoline could improve the therapeutic effect of NHIE,shorten the re-covery time of clinical manifestations,and improve nerve function,oxidative stress and vascular endothelial function without increasing drugs and adverse reactions.In addition,vascular endothelial function indexes are negatively correlated with NBNA score,which could be used as auxiliary reference indexes for judging NHIE.

OBJECTIVE To study the fingerprint of Denghong buyang huanwu granules （DBHG）， screen the quality markers， and establish the method for content determination of active ingredients. METHODS HPLC method was adopted. The fingerprints of 10 batches of DBHG （S1-S10） were established by using the Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine （2012 edition）， and similarity evaluation was also performed. Traditional Chinese medicine pieces attribution analysis， cluster analysis （CA）， principal component analysis （PCA） and orthogonal partial least squares discriminant analysis （OPLS-DA） were conducted for common peaks， and quality biomarkers were screened based on variable importance projection （VIP） values＞1. The contents of 10 batches of samples were determined by the same HPLC method， such as salidroside， tyrosol， calycosin-7-O-beta-D-glucoside， scutellarin and calycosin. RESULTS A total of 25 common peaks were obtained in the fingerprints for 10 batches of samples and 6 common peaks were identified， i.e. salidroside， tyrosol， paeoniflorin， calycosin-7-O-beta-D-glucoside， scutellarin， calycosin. Their similarities were greater than 0.9， and 10 common peaks of them were unique components of Erigeron breviscapus. DBHG could be clustered into 2 categories by using CA and PCA； S4-S5， and S7 could be clustered into one category and other samples could be clustered into one category. The corresponding components of peaks 16 （scutellarin）， 12， 15 （calycosin-7-O-beta-D-glucoside）， 13 （paeoniflorin）， and 14 were quality markers. The average contents of salidroside， tyrosol， calycosin-7-O-beta-D-glucoside， scutellarin and calycosin were 1.64， 0.45， 0.31， 0.73， 0.15 mg/g in 10 batches of samples. CONCLUSIONS HPLC fingerprint for DBHG and a method for determining the contents of five active ingredients including salidroside are successfully established. Five quality markers have been screened. It can be used for the quality control of the preparation.

Community-acquired respiratory infection is the commonest cause of sepsis presenting to emergency departments. Yet current experimental animal models simulate peritoneal sepsis with intraperitoneal (I.P.) injection of lipopolysaccharide (LPS) as the predominant route. We aimed to compare the progression of organ injury between I.P. LPS and intranasal (I.N.) LPS in order to establish a better endotoxemia murine model of respiratory sepsis. Eight weeks old male BALB/c mice received LPS-Escherichia coli doses at 0.15, 1, 10, 20, 40 and 100 mg per kg body weight (e.g. LPS-10 is a dose of 10 mg/kg body weight). Disease severity was monitored by a modified Mouse Clinical Assessment Score for Sepsis (M-CASS; range 0–21). A M-CASS score ≥ 10 or a weight reduction of ≥ 20%, was used as a criterion for euthanasia. The primary outcome was the survival rate (either no death or no need for euthanasia). The progression of disease was specified as M-CASS, body weight, blood glucose, histopathological changes to lung, liver, spleen, kidney, brain and heart tissues. Survival rate in I.P. LPS-20 mice was 0% (2/3 died; 1/3 euthanized with M-CASS > 10) at 24 h. Survival rate in all doses of I.N. LPS was 100% (20/20; 3–4 per group) at 96 h. 24 h mean M-CASS post-I.P. LPS-10 was 6.4/21 significantly higher than I.N. LPS-10 of 1.7/21 (Unpaired t test, P < 0.05). Organ injury was present at 96 h in the I.P. LPS-10 group: lung (3/3; 100%), spleen (3/3; 100%) and liver (1/3; 33%). At 24 h in the I.P. LPS-20 group, kidney injury was observed in the euthanized mouse. At 96 h in the post-I.N. LPS-20 group, only lung injury was observed in 2/3 (67%) mice (Kruskal-Wallis test with Dunn’s, P < 0.01). At 24 h in the post-I.N. LPS-100 group all (4/4) mice had evidence of lung injury. Variable doses of I.N. LPS in mice produced lung injury but did not produce sepsis. Higher doses of I.P. LPS induced multi-organ injury but not respiratory sepsis. Lethal models of respiratory virus, e.g., influenza A, might provide alternative avenues that can be explored in future research.

Objective: To evaluate the value of multi-detector CT (MDCT) upper airway imaging in the diagnosis of obstructive sleep apnea hypopnea syndrome ( OSAHS) and in determining the location of upper airway obstruction． Methods : MDCT was used to scan the upper airways of 85 clinically confirmed adult patients with different degrees of OSAHS (73 males and 12 females) in calm breathing phase and forced inhalation phase and 60 normal adults (50 males and 10 females) in calm breathing phase to obtain nasal cavity，nasopharynx，palatopharynx and oglosopharynx volumes．Parapharyngeal fat volume was measured in OSAHS patients and normal subjects．In addition，three groups of clinical data related to OSAHS patients were recorded，including sleep apnea hypopnea index (AHI) ，body mass index ( BMI) and lowest blood oxygen saturation ( LaSO2 ) ．Finally，the measured data and clinical data of each group were statistically analyzed. Results : The volume of nasopharynx and palatopharynx in the calm breathing group was significantly smaller than that in the control group，with statistical significance．Palatopharyngeal volume forced inspiratory phase was significantly smaller than calm breathing phase in the experimental group．The parapharyngeal fat volume in the experimental group was significantly higher than that in the control group．AHI was positively correlated with BMI and parapharyngeal fat volume．LaSO2 was negatively correlated with AHI and BMI，respectively． Conclusion MDCT upper airway imaging has good clinical application value in the diagnosis，treatment and postoperative evaluation of OSAHS disease due to the significant anatomical difference between OSAHS patients and normal subjects.

Objective To analyze the trend of suicide mortality among the elderly aged 60 years and above in Wuhan from 2014 to 2019, to understand the disease burden of suicide deaths among the elderly adults in Wuhan, and to provide a basis for decision making to carry out suicide interventions in the elderly population. Methods The data on suicide deaths in the elderly adults of Wuhan residents whose death age was 60 years or older were collected from 2014 to 2019 using the Wuhan City's Cause of Death Monitoring Information System. Mortality, standardized mortality, years of life lost (YLL) due to early death and average years of life lost (AYLL) were calculated separately. Statistical analysis was performed using SPSS 26.0. The χ² test was used to compare the suicide mortality rates among the elderly population by gender and region, and the annual percentage change (APC) was used for trend analysis. Results From 2014 to 2019, a total of 1010 suicide deaths were reported among elderly adults aged 60 years and older in Wuhan, with crude suicide mortality rates ranging from 7.60 to 10.77/100 000. The suicide mortality rate of elderly men was higher than that of elderly women. The suicide mortality rate of rural elderly adults was higher than that of urban elderly adults, and the suicide mortality rate of the rural elderly was decreasing. The overall suicide mortality rate of elderly people in Wuhan increased significantly with age, and the differences between the average suicide mortality rates of elderly males and elderly females in 2014-2019 were statistically significant among all age groups (P<0.01 or P<0.05). From 2014 to 2019, the YLL rate of suicide death among the elderly in Wuhan showed a trend of decreasing first and then increasing, and AYLL kept a slight fluctuation as a whole. The trends of both YLL rate and AYLL changes were not statistically significant. Conclusion The suicide mortality rate of elderly adults aged 60 years and above in Wuhan is high, especially in rural elderly men. The burden of disease caused by suicide deaths in the elderly is high, so it is necessary to take a variety of targeted measures to prevent and reduce the incidence of suicide among the elderly.

OBJECTIVE: To determine the carrier rate for common recessive genetic diseases in Chenzhou region in order to provide a reference for carrier screening in this region. METHODS: Targeted capture and high-throughput sequencing were carried out to detect potential variants of 79 genes associated with 88 recessive genetic diseases. Couples at risk were provided with prenatal diagnosis upon their subsequent pregnancies. RESULTS: A total of 1314 individuals were enrolled, among whom 355 (27.02%) were found to be carrier for at least one disease. The carrier rates for 8 diseases have exceeded 1%, with the most common two including thalassemia (11.72%, 154/1314) and autosomal recessive deafness (5.48%, 72/1314). Ten couples were found to be at risk for producing affected offspring. Among these, five females were carriers for X-linked recessive genetic diseases. Following genetic counseling, seven couples had accepted prenatal diagnosis, and 3 affected fetuses were diagnosed. CONCLUSION The disease types and pathogenic variants of Chenzhou region have differed from previously reported. Further research is required to validate the above finding with a larger populations.
Female ; Pregnancy ; Humans ; China ; Prenatal Diagnosis ; Fetus ; Genetic Counseling ; Genetic Diseases, X-Linked

Objective:To explore the genetic basis for a child featuring tetrahydrobiopterin deficiency and global developmental delay.Methods:Clinical and laboratory examinations were carried out for the child. Genomic DNA of the patient was subjected to high-throughput sequencing to identify genetic variants associated with hyperphenylalaninemia. Candidate variants were verified by Sanger sequencing.Results:The result of blood tandem mass spectrometry showed that the Phenylalanine in the blood was 642.7 μmol/l, and the ratio of Phenylalanine/Tyrosine was 5.42. Analysis of urinary pterin: neopterin 0.09 mmol/mol Cr, biopterin 0.04 mmol/mol Cr, biopterin% 77%, which suggested tetrahydrobiopterin deficiency. The parents of the proband were first cousins. DNA sequencing revealed that the proband has harbored homozygous c. 353A>T variants in exon 2 of the GCH1 gene, for which his great grandmother, grandfather, mother, uncle, father and elder brother were heterozygous carriers with normal phenotype and no clinical symptoms associated with dopa responsive dystonia. Conclusion:The homozygous c. 353A>T variant of the GCH1 gene probably underlay the tetrahydrobiopterin deficiency in this pedigree of consanguineous marriage.

Protein self-assemblies at the micro- and nano-scale are of great interest because of their morphological diversity and good biocompatibility. High-throughput screening of protein self-assembly at different scales and morphologies using protein crystallization screening conditions is an emerging method. When using this method to screen protein self-assembly conditions, some apparently transparent droplets are often observed, in which it is not clear whether self-assembly occurs. We explored the interaction between β-lactoglobulin and the protein crystallization kit Index™ C10 and observed the presence of micro- and nano-scale protein self-assemblies in the transparent droplets. The diverse morphology of the micro- and nano-scale self-assemblies in the transparent droplets formed by mixing different initial concentrations of β-lactoglobulin and Index™ C10 was further investigated by scanning electron microscope. Self-assembly process of fluorescence-labelled β-lactoglobulin was monitored continuously by laser confocal microscope, allowing real-time observation of the liquid-liquid phase separation phenomenon and the morphology of the final self-assemblies. The internal structure of the self-assemblies was gradually ordered over time by in-situ X-ray diffraction. This indicates that the self-assembly phenomenon within transparent droplets, observed in protein self-assembly condition screening experiments, is worthy of further in-depth exploration.
Crystallization ; Lactoglobulins

Objective:To investigate the changes and to establish a reference interval of bile acid profile of healthy pregnant women in the second and third trimesters of pregnancy in our hospital.Methods:A total of 298 healthy singleton pregnant women who underwent prenatal examination in the Department of Obstetrics of Zhejiang Provincial People′s Hospital from July 2019 to August 2020 were enrolled in this study. The overnight fasting serum samples were collected from all subjects during their second and third trimesters of pregnancy. The concentrations of 15 bile acids(cholic acid, chenodeoxycholic acid, deoxycholic acid, ursodeoxycholic acid, lithocholic acid and their glycine-and taurine-conjugated types)were analyzed by liquid chromatography tandem mass spectrometry (LC-MS/MS). The characteristics of changes were analyzed and the reference intervals were determined for the second and third trimesters. The concentrations of 15 bile acids and total bile acids were skewed-distributed, and 99 percentiles (P 99) were used to represent the unilateral upper limit of the reference interval. Results:There was significant difference in the serum levels of glycine cholic acid (GCA), taurocholic acid (TCA), glycine ursodeoxycholic acid (GUDCA) and lithocholic acid (LCA) between the second and third trimesters healthy pregnant females ( P<0.05). For other 11 bile acids, there was no significant difference. The levels of total bile acids, primary or secondary bile acids, free or conjugated bile acids (glycine-bound and taurine-bound bile acids) were stable with gestation. Conclusion:Primary, secondary or free, and conjugated bile acids in healthy pregnant women remained stable at T 2 and T 3, with significant differences in only a few subtypes of bile acids. While the correlation between glycine-bound and taurine-bound bile acids showed a weakening trend at T 3 ( P<0.05). It is necessary to establish reference intervals of bile acids for healthy pregnant women in this area. This study provided data support for future research on related diseases during pregnancy.