OBJECTIVE To construct novel central nervous system(CNS)-targeted lipid nanoparti-cles for the treatment of organophosphorus-induced brain injury in mice.METHODS(1)Preparation,screening,and characterization of lipid nanoparticles.① Lipid nanoreactivators were prepared using the thin-film hydration method,with asoxime(HI-6)as the therapeutic drug and lipid carriers composed of 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-L-serine(POPS),1,2-dipalmitoyl-sn-glycero-3-phospho-choline(DPPC),and cholesterol(CHOL)(PDC)at varying molar ratios(1∶6∶3,3∶4∶3,5∶2∶3 and 7∶0∶3)(HI-6@PDC 1∶6∶3,3∶4∶3,5∶2∶3 and 7∶0∶3).② FLU-labeled lipid nanocarriers(FLU@PDC 1∶6∶3,3∶4∶3,5∶2∶3,and 7∶0∶3)were prepared and physically mixed with phospholipase A2(PLA2)solution(at the final PLA2 concentration of 10 kU·L-1)to obtain FLU@PDC+PLA2.Male KM mice were randomly divided into normal control(PBS),FLU,and FLU@PDC+PLA2(1∶6∶3,3∶4∶3,5∶2∶3,and 7∶0∶3)groups(n=7 per group).After intravenous(iv)administration(FLU dose:1 mg·kg-1,carrier dose:80 mg·kg-1),brain tissues were collected at 1 h,homogenized,centrifuged,and analyzed via fluorescence spectrophotom-etry to screen the optimal CNS-targeted lipid carrier composition.③ The morphology of HI-6@PDC 5∶2∶3 was characterized by transmission electron microscope(TEM).The particle size,polydispersity index(PDI),and zeta potential of HI-6@PDC 5∶2∶3 were measured using a Zeta potential and particle size analyzer.Encapsulation efficiency and loading efficiency of HI-6@PDC 5∶2∶3 were determined using an ultrafiltration centrifugation method combined with high-performance liquid chromatography(HPLC).In vitro release kinetics of HI-6@PDC 5∶2∶3 and HI-6@PDC+PLA2 5∶2∶3 were assessed using a dialysis bag diffusion method combined with fluorescence spectrophotometry.(2)Validation of CNS targeting.① Cyanine7(Cy7)-labeled PDC 5∶2∶3(Cy7@PDC)was prepared and mixed with PLA2 solution(Cy7@PDC+PLA2 5∶2∶3).Mice were divided into normal control,Cy7,Cy7@PDC 5∶2∶3 and Cy7@PDC+PLA2 5∶2∶3 groups(n=3 per group).After iv injection(Cy7 dose:1 mg·kg-1,carrier dose:80 mg·kg-1),brain fluorescence was visualized at 3 h using a small animal in vivo imaging(IVIS)system.② Cyanine 3(Cy3)-labeled PDC 5∶2∶3(Cy3@PDC 5∶2∶3)was prepared and mixed with PLA2 solution(Cy3@PDC+PLA2 5∶2∶3).Mice were divided into Cy3@PDC 5∶2∶3 and Cy3@PDC+PLA2 5∶2∶3 groups(n=3 per group).After iv injection(Cy3 dose:1 mg·kg-1,carrier dose:80 mg·kg-1),brain tissues were collected at 2 h for fluorescent staining and Cy3 fluorescence observation.(3)Therapeutic efficacy eval-uation.① Male KM mice were randomly divided into normal control,brain injury,HI-6 treatment,and HI-6@PDC+PLA2 5∶2∶3 treatment groups(n=6 per group).Except for the normal control,all the mice were subcutaneously(sc)injected with soman(120 μg·kg-1),followed by immediate iv treatment(HI-6 dose:22 mg·kg-1,carrier dose:80 mg·kg-1).At 10 min,orbital blood and brain tissues were collected before brain weight was recorded.Acetylcholinesterase(AChE)reactivation in blood and brain was measured using the Ellman method.② Grouping and treatment were identical to ①(n=3 per group).At 24 h,brain tissues were collected for HE staining to assess histopathological damage.③ Mice were divided into brain injury and HI-6@PDC+PLA2 5∶2∶3 treatment groups(n=10 per group)and treated as in ①(soman dose:220 ug·kg-1).Survival rates,neurotoxic symptoms(tremors,salivation),and seizure latency were recorded,and survival curves were plotted.RESULTS(1)PDC 5∶2∶3 exhibited the highest brain fluorescence,indicating optimal CNS targeting.HI-6@PDC 5∶2∶3 appeared in regular spherical shapes,and were negatively charged,with a size of(219.4±3.1)nm,PDI of 0.4±0.02,entrapment effi-ciency of 72.9％and loading efficiency of 49.7％.HI-6@PDC+PLA2 5∶2∶3 showed a cumulative release of 43.5％at 60 min,which was lower than that of rhodamine B(RB)but sufficient for CNS therapeutic timelines.(2)In vivo fluorescence and pathological fluorescence confirmed PLA2-mediated CNS delivery.(3)HI-6@PDC+PLA2 5∶2∶3 significantly enhanced AChE reactivation in the blood and brain compared to HI-6.Histopathology revealed mitigated brain injury in treated mice.HI-6@PDC+PLA2 5∶2∶3 prolonged survival,reduced convulsions,alleviated neurotoxicity,and extended seizure latency.CONCLUSION HI-6@PDC 5∶2∶3 can effectively cross the blood-brain barrier via PLA2 mediation,demonstrating strong CNS targeting.It can significantly improve AChE reactivation in peripheral and central tissues and offers potent therapeutic efficacy against organophosphate-induced brain injury.

OBJECTIVE To construct novel central nervous system(CNS)-targeted lipid nanoparti-cles for the treatment of organophosphorus-induced brain injury in mice.METHODS(1)Preparation,screening,and characterization of lipid nanoparticles.① Lipid nanoreactivators were prepared using the thin-film hydration method,with asoxime(HI-6)as the therapeutic drug and lipid carriers composed of 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-L-serine(POPS),1,2-dipalmitoyl-sn-glycero-3-phospho-choline(DPPC),and cholesterol(CHOL)(PDC)at varying molar ratios(1∶6∶3,3∶4∶3,5∶2∶3 and 7∶0∶3)(HI-6@PDC 1∶6∶3,3∶4∶3,5∶2∶3 and 7∶0∶3).② FLU-labeled lipid nanocarriers(FLU@PDC 1∶6∶3,3∶4∶3,5∶2∶3,and 7∶0∶3)were prepared and physically mixed with phospholipase A2(PLA2)solution(at the final PLA2 concentration of 10 kU·L-1)to obtain FLU@PDC+PLA2.Male KM mice were randomly divided into normal control(PBS),FLU,and FLU@PDC+PLA2(1∶6∶3,3∶4∶3,5∶2∶3,and 7∶0∶3)groups(n=7 per group).After intravenous(iv)administration(FLU dose:1 mg·kg-1,carrier dose:80 mg·kg-1),brain tissues were collected at 1 h,homogenized,centrifuged,and analyzed via fluorescence spectrophotom-etry to screen the optimal CNS-targeted lipid carrier composition.③ The morphology of HI-6@PDC 5∶2∶3 was characterized by transmission electron microscope(TEM).The particle size,polydispersity index(PDI),and zeta potential of HI-6@PDC 5∶2∶3 were measured using a Zeta potential and particle size analyzer.Encapsulation efficiency and loading efficiency of HI-6@PDC 5∶2∶3 were determined using an ultrafiltration centrifugation method combined with high-performance liquid chromatography(HPLC).In vitro release kinetics of HI-6@PDC 5∶2∶3 and HI-6@PDC+PLA2 5∶2∶3 were assessed using a dialysis bag diffusion method combined with fluorescence spectrophotometry.(2)Validation of CNS targeting.① Cyanine7(Cy7)-labeled PDC 5∶2∶3(Cy7@PDC)was prepared and mixed with PLA2 solution(Cy7@PDC+PLA2 5∶2∶3).Mice were divided into normal control,Cy7,Cy7@PDC 5∶2∶3 and Cy7@PDC+PLA2 5∶2∶3 groups(n=3 per group).After iv injection(Cy7 dose:1 mg·kg-1,carrier dose:80 mg·kg-1),brain fluorescence was visualized at 3 h using a small animal in vivo imaging(IVIS)system.② Cyanine 3(Cy3)-labeled PDC 5∶2∶3(Cy3@PDC 5∶2∶3)was prepared and mixed with PLA2 solution(Cy3@PDC+PLA2 5∶2∶3).Mice were divided into Cy3@PDC 5∶2∶3 and Cy3@PDC+PLA2 5∶2∶3 groups(n=3 per group).After iv injection(Cy3 dose:1 mg·kg-1,carrier dose:80 mg·kg-1),brain tissues were collected at 2 h for fluorescent staining and Cy3 fluorescence observation.(3)Therapeutic efficacy eval-uation.① Male KM mice were randomly divided into normal control,brain injury,HI-6 treatment,and HI-6@PDC+PLA2 5∶2∶3 treatment groups(n=6 per group).Except for the normal control,all the mice were subcutaneously(sc)injected with soman(120 μg·kg-1),followed by immediate iv treatment(HI-6 dose:22 mg·kg-1,carrier dose:80 mg·kg-1).At 10 min,orbital blood and brain tissues were collected before brain weight was recorded.Acetylcholinesterase(AChE)reactivation in blood and brain was measured using the Ellman method.② Grouping and treatment were identical to ①(n=3 per group).At 24 h,brain tissues were collected for HE staining to assess histopathological damage.③ Mice were divided into brain injury and HI-6@PDC+PLA2 5∶2∶3 treatment groups(n=10 per group)and treated as in ①(soman dose:220 ug·kg-1).Survival rates,neurotoxic symptoms(tremors,salivation),and seizure latency were recorded,and survival curves were plotted.RESULTS(1)PDC 5∶2∶3 exhibited the highest brain fluorescence,indicating optimal CNS targeting.HI-6@PDC 5∶2∶3 appeared in regular spherical shapes,and were negatively charged,with a size of(219.4±3.1)nm,PDI of 0.4±0.02,entrapment effi-ciency of 72.9％and loading efficiency of 49.7％.HI-6@PDC+PLA2 5∶2∶3 showed a cumulative release of 43.5％at 60 min,which was lower than that of rhodamine B(RB)but sufficient for CNS therapeutic timelines.(2)In vivo fluorescence and pathological fluorescence confirmed PLA2-mediated CNS delivery.(3)HI-6@PDC+PLA2 5∶2∶3 significantly enhanced AChE reactivation in the blood and brain compared to HI-6.Histopathology revealed mitigated brain injury in treated mice.HI-6@PDC+PLA2 5∶2∶3 prolonged survival,reduced convulsions,alleviated neurotoxicity,and extended seizure latency.CONCLUSION HI-6@PDC 5∶2∶3 can effectively cross the blood-brain barrier via PLA2 mediation,demonstrating strong CNS targeting.It can significantly improve AChE reactivation in peripheral and central tissues and offers potent therapeutic efficacy against organophosphate-induced brain injury.

Objective To analyze the temporal changing trend of postoperative pneumonia(POP)monitoring data in a tertiary first-class cancer hospital in Fujian Province from 2018 to 2023,and provide reference for the effective-ness of implementation of healthcare-associated infection(HAI)prevention and control measures.Methods The temporal changing trend of POP monitoring data of cancer patients in this hospital from 2018 to 2023 was analyzed by Joinpoint regression model,and the average annual percentage change(AAPC)was calculated.Results From 2018 to 2023,the POP incidences of all cancer patients and patients with different tumors in this hospital were as follows:3.46％in all cancer patients,4.77％,18.16％,11.50％,4.66％,0.85％,3.74％,and 0.46％in pa-tients with lung cancer,esophageal cancer,gastric cancer,intestinal cancer,gynecological tumors,hepatobiliary-pancreatic tumor,as well as head and neck tumors,respectively.From 2018 to 2023,the POP incidence of all can-cer patients in the hospital decreased from 5.47％to 1.73％,and POP incidences of patients with lung cancer,gas-tric cancer,and intestinal cancer decreased from 12.23％,14.93％,and 4.40％to 2.60％,3.73％,and 2.09％,respectively.Joinpoint regression model analysis showed that from 2018 to 2023,the AAPC of POP incidence of all cancer patients in the hospital was-19.78％,and the AAPCs of patients with lung cancer,gastric cancer,and in-testinal cancer were-23.69％,-27.30％,and-19.40％,respectively.The incidences of POP in all cancer pa-tients,as well as patients with lung cancer,gastric cancer,and intestinal cancer all showed downward trends,and the differences were all statistically significant(all P＜0.05).According to age,the AAPCs of the ≤60 and＞60 year old groups were-22.02％and-20.48％,respectively,both groups showed statistically significant difference in trends(both P＜0.05).In terms of gender,the AAPCs of the male and female groups were-16.56％and-28.35％,respectively,both groups showed statistically significant difference in trends(both P＜0.05).From 2018 to 2023,Klebsiella pneumoniae showed a significant upward trend in the constituent of POP pathogens in cancer patients,with an AAPC of 6.92％,and the difference was statistically significant(P＜0.05).Conclusion The incidences of POP in some cancer patients in the hospital present significant downward trends,indicating that HAI infection prevention and control measures are effective,but it is still necessary to strengthen the meticulous management of the whole perioperative process.

Objective To analyze the temporal changing trend of postoperative pneumonia(POP)monitoring data in a tertiary first-class cancer hospital in Fujian Province from 2018 to 2023,and provide reference for the effective-ness of implementation of healthcare-associated infection(HAI)prevention and control measures.Methods The temporal changing trend of POP monitoring data of cancer patients in this hospital from 2018 to 2023 was analyzed by Joinpoint regression model,and the average annual percentage change(AAPC)was calculated.Results From 2018 to 2023,the POP incidences of all cancer patients and patients with different tumors in this hospital were as follows:3.46％in all cancer patients,4.77％,18.16％,11.50％,4.66％,0.85％,3.74％,and 0.46％in pa-tients with lung cancer,esophageal cancer,gastric cancer,intestinal cancer,gynecological tumors,hepatobiliary-pancreatic tumor,as well as head and neck tumors,respectively.From 2018 to 2023,the POP incidence of all can-cer patients in the hospital decreased from 5.47％to 1.73％,and POP incidences of patients with lung cancer,gas-tric cancer,and intestinal cancer decreased from 12.23％,14.93％,and 4.40％to 2.60％,3.73％,and 2.09％,respectively.Joinpoint regression model analysis showed that from 2018 to 2023,the AAPC of POP incidence of all cancer patients in the hospital was-19.78％,and the AAPCs of patients with lung cancer,gastric cancer,and in-testinal cancer were-23.69％,-27.30％,and-19.40％,respectively.The incidences of POP in all cancer pa-tients,as well as patients with lung cancer,gastric cancer,and intestinal cancer all showed downward trends,and the differences were all statistically significant(all P＜0.05).According to age,the AAPCs of the ≤60 and＞60 year old groups were-22.02％and-20.48％,respectively,both groups showed statistically significant difference in trends(both P＜0.05).In terms of gender,the AAPCs of the male and female groups were-16.56％and-28.35％,respectively,both groups showed statistically significant difference in trends(both P＜0.05).From 2018 to 2023,Klebsiella pneumoniae showed a significant upward trend in the constituent of POP pathogens in cancer patients,with an AAPC of 6.92％,and the difference was statistically significant(P＜0.05).Conclusion The incidences of POP in some cancer patients in the hospital present significant downward trends,indicating that HAI infection prevention and control measures are effective,but it is still necessary to strengthen the meticulous management of the whole perioperative process.

This study presents prenatal diagnosis and genetic analysis of a pedigree with X-linked intellectual disability. A gravida at approximately 20 gestational weeks underwent prenatal diagnosis following non-invasive prenatal testing suggested sex chromosome abnormalities. Copy number variation (CNV) sequencing identified a 5.7 Mb duplication at Xp22.2p22.11 in the fetus, which initially classified as a variant of uncertain clinical significance. This duplication was inherited from the phenotypically normal mother, while paternal CNV results were normal. Genetic testing of four intellectually disabled family members revealed the identical 5.7 Mb duplication. Through expanded pedigree analysis, the pathogenicity classification of the Xp22.2p22.11 microduplication was upgraded to likely pathogenic. After comprehensive genetic counseling, the family elected pregnancy termination with informed consent.

This study presents prenatal diagnosis and genetic analysis of a pedigree with X-linked intellectual disability. A gravida at approximately 20 gestational weeks underwent prenatal diagnosis following non-invasive prenatal testing suggested sex chromosome abnormalities. Copy number variation (CNV) sequencing identified a 5.7 Mb duplication at Xp22.2p22.11 in the fetus, which initially classified as a variant of uncertain clinical significance. This duplication was inherited from the phenotypically normal mother, while paternal CNV results were normal. Genetic testing of four intellectually disabled family members revealed the identical 5.7 Mb duplication. Through expanded pedigree analysis, the pathogenicity classification of the Xp22.2p22.11 microduplication was upgraded to likely pathogenic. After comprehensive genetic counseling, the family elected pregnancy termination with informed consent.

Objective To investigate the age-related changes of biomechanical properties for humerus, femur and tibia in male rats and their application values in age estimation. Methods According to different weeks of age, 90 healthy male SD rats were divided into 2, 4, 6, 8, 17, 26, 52, 78 and 104-week groups with 10 rats in eachgroup. After the rats were executed by excessive anesthesia, humerus, femur, and tibia were separated and the attached soft tissues were removed. The length of the above-mentioned bones and the diameter of the middle section (compression site) were measured with vernier caliper, and the three-point bending test was conducted with electronic universal material testing machine to detect the ultimate load and displacement under ultimate load. Results There were significant differences in the ultimate load of humerus, femur and tibia among male rats in different age groups (P<0. 05). With the increase of week age, the ultimate loads of the humerus, femur and tibia increased first and then decreased, and reached the peak value in 52-week age group, showing a strong positive correlation with week age before 52 weeks ( r = 0. 884,0. 933,0. 929, P<0. 05). There was no significant difference in humerus and tibia. The displacement of femur under ultimate load was weakly positively correlated with week age (R= 0. 406,P<0. 05). The age prediction accuracy for automatic linear modeling of ultimate load for humerus, femur, tibia and three above-mentioned bones in rats before 52-week age was 78. 2% , 86. 8% , 84. 1% and 88. 3% , respectively. There was a strong positive correlation between the length of humerus, femur and tibia and the ultimate load (R= 0. 904, 0. 897, 0. 814, P<0. 05). The diameters of humerus, femur and tibia were strongly positively correlated with the ultimate load (R = 0. 759, 0. 814 and 0. 745, P<0. 05). Conclusions The ultimate loads of humerus, femur and tibia in male rats increased first and then decreased with age, and were positively correlated with age before 52 weeks, which could be used for age inference. The highest accuracy of age estimation was ultimate loads of three bones, followed by femur. The length/ middle diameter of humerus, femur and tibia were strongly positively correlated with the ultimate load.

Objective To investigate the effects of oral care using hydrogen peroxide and sodium bicarbonate to prevent neonatal ventilator associated pneumonia(VAP). Methods Totally 209 neonates were recruited and divided into the experimental group with 104 cases and the control group with 105 cases by using random number table method. Based on conventional mechanical ventilation nursing,the experimental group received oral care using 1.5%hydrogen peroxide combined with 2.5% sodium bicarbonate,Q8H,while the control group received oral care using only 2.5% sodium bicarbonate,Q8H. Positive results of bacteria detection in tracheal sputum culture,the incidence rate of VAP,mechanical ventilation time,hospitalization time and hospitalization costs were compared between two groups. Results After 48 hours of mechanical ventilation,the difference in positive results of bacteria detection in tracheal sputum culture between two groups was statistically significant(P<0.05). The difference of incidence rate of VAP between two groups showed no statistical significance(P>0.05) when the duration of the mechanical ventilation was 48 hours. While after 48 hours of the mechanical ventilation,the difference of the incidence rate of VAP between two groups was statistically significant(P<0.05). The differences in mechanical ventilation time and hospitalization time between two groups were statistically significant(P<0.05). The hospitalization costs of the experimental group was higher than that of the control group,while the difference showed no statistical significance(P>0.05). Conclusion The combined usage of hydrogen peroxide and sodium bicarbonate for oral care can effectively eliminate neonatal oral bacteria colonization and prevent neonatal VAP,so as to reduce the time of mechanical ventilation and hospitaliza-tion, and decrease hospitalization costs.

Purpose To investigate the feasibility to diagnose and evaluate the vulnerability of the atherosclerotic plaque through contrast-enhanced ultrasonography. Materials and Methods Thirty-four patients with carotid atherosclerosis were enrolled in the study to take contrast-enhanced ultrasonography and were observed on whether the plaque was enhanced and the features of its enhancement. The peak intensity (PI), time to peak (TTP), and density echo (DE) were calculated according to the time-intensity curve with QLAB software. Results The ratio of enhanced malacoplakia and the mixed plaque showed no difference (χ2=0.847, P>0.05). The percentages of enhanced plaque distributed on the base, tail and the shoulder were 70.0%, 23.3% and 6.7%, respectively with significant difference (χ2=29.100, P<0.001); the distribution ratio of enhanced plaque from high to low were as follows: the plaque on the shoulder > the plaque on the base> the plaque on the tail. It was positively correlated between enhanced plaque and its distribution (r=0.404, P<0.01). The TTP of the ROI between the malacoplakias and the mixed plaques showed no difference (t=0.479, P>0.05). The PI and DE of the ROI in the malacoplakia and the mixed plaques were analyzed by the time-intensity curve and the differences proved to be statistically significant (t=7.497 and 12.224, P<0.05). Conclusion Contrast-enhanced ultrasonography could present the neovessels in the atherosclerotic plaque, which is helpful in evaluating the vulnerability of atherosclerotic plaque.

OBJECTIVETo investigate the feature of phenylalanine hydroxylase (PAH) gene mutations and provide guidance for genetic and prenatal diagnosis of patients with phenylketonuria from Shaanxi.

METHODSFor 55 patients whose blood Phe concentration was over 2.0 mg/dL, potential mutations in 13 exons and flanking sequences of the PAH gene were detected by PCR and DNA sequencing.

RESULTSA total of 98 mutations were detected in 110 PAH alleles, with the detection rate being 89.10%. Nine mutations have been identified in exon 7, which accounted for 33.67% of all. Exon 12 (14.29%) and exon 3 (12.24%) have followed. Thirty eight mutations, locating in exon2-exon12 and the flanking sequence, were detected in the 55 PKU patients. p.R243Q (24.49%) was the commonest mutation, whilstp.A47E, p.I65S and p.A259T were first discovered in China. After querying international databases including PAHdb and HGMD, the p.C334X was verified as the novel PAH gene mutation.

CONCLUSIONThe mutation spectrum of the PAH gene in Shaanxi has been identified. And a novel mutation has been identified. This may facilitate the diagnosis of PKU in the future.

Alleles ; Base Sequence ; Child ; Child, Preschool ; China ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Mutation ; Phenylalanine Hydroxylase ; blood ; genetics ; Phenylketonurias ; enzymology ; genetics