Objective To investigate the effect and mechanism of leucine zipper/EF-hand-containing transmembrane protein 1(LETM1)on proliferation,migration,apoptosis,osteogenic differentiation,and tumorigenesis in vivo of human osteosarcoma MG63 and 143B cells.Methods The osteosarcoma MG63 and 143B cells were divided into blank control group(without adenovirus infection),negative control group(sh-NC group,infected with RNAi negative control virus),and LETM1 knockdown group(sh-LETM1 group,infected with sh-LETM1 adenovirus).Western blotting was performed to detect LETM1 expression in normal human osteoblasts hFOB1.19 and osteosarcoma cells,and to verify the knockdown effect of adenovirus;cell clone formation assays and CCK-8 method were used to detect the proliferation of MG63 and 143B cells;wound-healing assay and Transwell assay were used to test cell migration;DAPI staining and Annexin V-APC/7-AAD flow cytometry double staining were used to detect the apoptosis of MG63 and 143B cells;alkaline phosphatase(ALP)staining and Alizarin Red S staining were used to evaluate early and late osteogenic differentiation of MG63 and 143B cells.Ten nude mice were divided into sh-NC group(n=5,injected subcutaneously into nude mice with 143B cells infected with RNAi negative control virus)and sh-LETM1 group(n=5,injected subcutaneously into nude mice with 143B cells infected with sh-LETM1 adenovirus),and nude mice subcutaneous tumor formation assay was used to examine the in vivo tumor-forming ability of 143B cells in each group.Results Western blotting showed that the expression of LETM1 protein in osteosarcoma MG63 and 143B cells was significantly higher than that in human normal osteoblasts hFOB1.19(P<0.05),and that the expression of LETM1 protein was markedly reduced after injection with sh-LETM1 adenovirus in MG63 and 143B cells.The results of cell clone formation assay and CCK-8 assay indicated that in MG63 and 143B osteosarcoma cells,the clone formation ability and proliferation ability were significantly reduced in sh-LETM1 group compared with sh-NC group and blank control group(P<0.01).The results of wound-healing assay and Transwell assay demonstrated that in MG63 and 143B osteosarcoma cells,the cell migration rate in sh-LETM1 group was significantly lower than that in sh-NC group and blank control group(P<0.01).DAPI staining and flow cytometry results revealed that the apoptosis rate in sh-LETM1 group was significantly higher than those in MG63 and 143B osteosarcoma cells in sh-NC group(P<0.01).Alkaline phosphatase staining and Alizarin red S staining experiments showed more stained areas and calcium salt nodules in MG63 and 143B osteosarcoma cells in sh-LETM1 group than those in sh-NC group and blank control group.The results of the subcutaneous tumor formation assay in nude mice indicated that subcutaneous tumor formation ability was reduced in 143B sh-LETM1 group compared with 143B sh-NC group.Conclusion LETM1 promotes the proliferation,migration and in vivo tumor formation of MG63 and 143B osteosarcoma cells and the mechanism may be related to the inhibition of apoptosis and osteogenic differentiation.

Humans ; Parkinson Disease/diagnosis* ; Machine Learning

Objective: To analyze the clinical epidemiological characteristics including composition of pathogens , clinical characteristics, and disease prognosis acute bacterial meningitis (ABM) in Chinese children. Methods: A retrospective analysis was performed on the clinical and laboratory data of 1 610 children <15 years of age with ABM in 33 tertiary hospitals in China from January 2019 to December 2020. Patients were divided into different groups according to age,<28 days group, 28 days to <3 months group, 3 months to <1 year group, 1-<5 years of age group, 5-<15 years of age group; etiology confirmed group and clinically diagnosed group according to etiology diagnosis. Non-numeric variables were analyzed with the Chi-square test or Fisher's exact test, while non-normal distrituction numeric variables were compared with nonparametric test. Results: Among 1 610 children with ABM, 955 were male and 650 were female (5 cases were not provided with gender information), and the age of onset was 1.5 (0.5, 5.5) months. There were 588 cases age from <28 days, 462 cases age from 28 days to <3 months, 302 cases age from 3 months to <1 year of age group, 156 cases in the 1-<5 years of age and 101 cases in the 5-<15 years of age. The detection rates were 38.8% (95/245) and 31.5% (70/222) of Escherichia coli and 27.8% (68/245) and 35.1% (78/222) of Streptococcus agalactiae in infants younger than 28 days of age and 28 days to 3 months of age; the detection rates of Streptococcus pneumonia, Escherichia coli, and Streptococcus agalactiae were 34.3% (61/178), 14.0% (25/178) and 13.5% (24/178) in the 3 months of age to <1 year of age group; the dominant pathogens were Streptococcus pneumoniae and the detection rate were 67.9% (74/109) and 44.4% (16/36) in the 1-<5 years of age and 5-<15 years of age . There were 9.7% (19/195) strains of Escherichia coli producing ultra-broad-spectrum β-lactamases. The positive rates of cerebrospinal fluid (CSF) culture and blood culture were 32.2% (515/1 598) and 25.0% (400/1 598), while 38.2% (126/330)and 25.3% (21/83) in CSF metagenomics next generation sequencing and Streptococcus pneumoniae antigen detection. There were 4.3% (32/790) cases of which CSF white blood cell counts were normal in etiology confirmed group. Among 1 610 children with ABM, main intracranial imaging complications were subdural effusion and (or) empyema in 349 cases (21.7%), hydrocephalus in 233 cases (14.5%), brain abscess in 178 cases (11.1%), and other cerebrovascular diseases, including encephalomalacia, cerebral infarction, and encephalatrophy, in 174 cases (10.8%). Among the 166 cases (10.3%) with unfavorable outcome, 32 cases (2.0%) died among whom 24 cases died before 1 year of age, and 37 cases (2.3%) had recurrence among whom 25 cases had recurrence within 3 weeks. The incidences of subdural effusion and (or) empyema, brain abscess and ependymitis in the etiology confirmed group were significantly higher than those in the clinically diagnosed group (26.2% (207/790) vs. 17.3% (142/820), 13.0% (103/790) vs. 9.1% (75/820), 4.6% (36/790) vs. 2.7% (22/820), χ²=18.71, 6.20, 4.07, all P<0.05), but there was no significant difference in the unfavorable outcomes, mortility, and recurrence between these 2 groups (all P>0.05). Conclusions: The onset age of ABM in children is usually within 1 year of age, especially <3 months. The common pathogens in infants <3 months of age are Escherichia coli and Streptococcus agalactiae, and the dominant pathogen in infant ≥3 months is Streptococcus pneumoniae. Subdural effusion and (or) empyema and hydrocephalus are common complications. ABM should not be excluded even if CSF white blood cell counts is within normal range. Standardized bacteriological examination should be paid more attention to increase the pathogenic detection rate. Non-culture CSF detection methods may facilitate the pathogenic diagnosis.
Adolescent ; Brain Abscess ; Child ; Child, Preschool ; Escherichia coli ; Female ; Humans ; Hydrocephalus ; Infant ; Infant, Newborn ; Male ; Meningitis, Bacterial/epidemiology* ; Retrospective Studies ; Streptococcus agalactiae ; Streptococcus pneumoniae ; Subdural Effusion ; beta-Lactamases

【Objective】 To compare the ability of body mass index （BMI）， body fat percentage （BFP）， waist circumference （WC）， waist-to-height ratio （WHtR）， visceral fat index （VFI） and the combinations of two kinds of obesity indices to predict the risk of hypertension. 【Methods】 Data collected in the baseline survey of “Gansu Province’s Urban and Rural Natural Population Cohort Establishment and Tumor Follow-up Study” were analyzed. Area under the curve （AUC） of ROC curve with covariates was used to analyze and compare the effects of individual obesity evaluation index and the combination of two kinds of obesity indices in predicting the risk of hypertension. 【Results】 Analyses of data of 20，079 adults showed that the AUC of BMI， WC， WHtR， BFP and VFI was 0.636， 0.604， 0.615， 0.614 and 0.619， respectively. AUC of the combination of BMI and WC （0.643） was higher than that of BMI （0.636）； however， the change rate of AUC was only 1.09%. AUC of the combinations of WC， WHtR and VFI， the three central obesity evaluation indices， and BFP， a general obesity evaluation index， were lower than that of BMI. The optimal cutoff value for BMI was 24.2 kg/m². 【Conclusion】 The effect of BMI in predicting the risk of hypertension is better than that of BFP， WC， WHtR and VFI. The effects of the combinations of the two kinds of obesity evaluation indices are not better than that of BMI. To prevent and control hypertension， adults should keep their BMI under overweight.

As the pipelines of nutrient and oxygen in the brain, cerebral vessels play an important role in the development and normal function of the brain. The development of the cerebrovascular system is divided into two processes, primitive vasculogenesis and angiogenesis. In this review, we summarize the roles, regulation mechanisms, as well as the interactions of the essential signaling molecules, such as the family members of vascular endothelial growth factor（VEGF）, neuropillin, Wnt, Notch, Hedgehog, in the vessel development, aiming to provide a reference for the basic and clinical investigation and the prevention and treatment of vascular diseases. Reset.

Traumatic brain injury (TBI) triggers the activation of the endogenous coagulation mechanism, and a large amount of thrombin is released to curb uncontrollable bleeding through thrombin receptors, also known as protease-activated receptors (PARs). However, thrombin is one of the most critical factors in secondary brain injury. Thus, the PARs may be effective targets against hemorrhagic brain injury. Since the PAR1 antagonist has an increased bleeding risk in clinical practice, PAR4 blockade has been suggested as a more promising treatment. Here, we explored the expression pattern of PAR4 in the brain of mice after TBI, and explored the effect and possible mechanism of BMS-986120 (BMS), a novel selective and reversible PAR4 antagonist on secondary brain injury. Treatment with BMS protected against TBI in mice. mRNA-seq analysis, Western blot, and qRT-PCR verification in vitro showed that BMS significantly inhibited thrombin-induced inflammation in astrocytes, and suggested that the Tab2/ERK/NF-κB signaling pathway plays a key role in this process. Our findings provide reliable evidence that blocking PAR4 is a safe and effective intervention for TBI, and suggest that BMS has a potential clinical application in the management of TBI.

Objective: To investigate the clinical value of independently-designed three-demensional printed navigation templates for hepatectomy in hepatic surgical oncology. Methods: A retrospective study was conducted on 45 patients (including 36 males and 9 females, aged from 40 to 70 years) with hepatocelluar carcinoma treated by hepatoctomy from Department of Hepatobiliary and Intestinal Surgery of Hunan Cancer Hospital from June 2016 to June 2018. These patients were divided into observation group (patients conducted with the 3D navigation-template-aided operation, n=25) and control group (patients conducted with regular operation, n=20) according to operation approach. Virtual liver resection volume and actual liver resection volume were calculated in the observation group, and the correlationship between the volume of virtual liver resection and actual liver resection was analyzed by Pearson correlation analysis. The operative time, intraoperative blood loss, intraoperative intrahepatic duct injury and postoperative hospital stay were compared between the two groups. Results: In the observation group, 3D modeling was successfully completed and intrahepatic and extrahepatic duct reconstruction was successful, surgical simulation. There were no statistically significant difference between the virtual and actual resected liver volume in the observation group (P>0.05). The virtual resected liver volume was positively correlated with the actual resected liver volume (r=0.991, P<0.05). Volume of intraoperative blood loss in the observation group was lower than that in the control group [(245.1±22.3) ml vs. (308.6±19.9) ml], and the length of hospital stay was shorter than that in the control group [(7.4±0.2) d vs. (8.6±0.4) d], the difference was statistically significant (P<0.05). There was no accidental injury of intrahepatic duct in the observation group, 1 case of accidental injury of middle hepatic vein and 1 case of accidental injury of secondary branch of right hepatic vein in the control group. The AST of the observation group was significantly reduced compared with that of the control group on the 4 th and 7th days after the operation, and the difference was statistically significant (P<0.05). Conclusions 3D reconstruction of liver based on high quality MR images, can facilitate restoring the adjoint relationship between hepatic duct system and liver tumor. 3D printed navigation templates effectively guide certain kinds of hepatectomy, ensure that the cutting edge at the same time protect the rest of the liver tissue, reduce bleeding and accidental damage, realize accurate tumor resection.

Objective To evaluate the value of serum aminoterminal pro-brain natriuretic peptide (NT-proBNP) and interleukin(IL)-6 levels in diagnosis and severity assessment of the preterm infants with respiratory distress syndrome(RDS).Methods Totally 150 preterm infants with RDS who were hospitalized in our center from August 2016 to March 2018 were enrolled in this study as the RDS group. These infants were further divided into grades 1,2,3,and 4 according to chest radiography. In addition,158 preterm infants without RDS hospitalized in our center during the same period were included as the controls (control group). Serum NT-proBNP and IL-6 levels were measured by ELISA on days 1,3,and 7 after birth,and their pulmonary arterial pressure (PAP) was monitored as well.Results Serum NT-proBNP and IL-6 levels in RDS group were significantly higher than those in control group on day 1 (t=-91.04,P=0.000;t=-11.03,P=0.000),day 3 (t=-89.10,P=0.000;t=-9.909,P=0.000),and day 7 (t=-87.91,P=0.000;t=-8.548,P=0.000). There were significant differences in NT-proBNP levels among grades 1,2,3,and 4 on day 1 (F=50.89,P=0.000),day 3 (F=49.16,P=0.000),and day 7 (F=45.45,P=0.000),showing an increasing trend. Serum IL-6 levels showed no significant difference among grades 1,2,3,and 4 on day 1 (F=0.89,P=0.448),day 3 (F=0.76,P=0.518),and day 7 (F=0.85,P=0.469). The PAP of the RDS group on days 1,3,and 7 was (49.3±3.7),(40.1±5.4),and (39.0±2.6)mmHg (1 mmHg=0.133 kPa),which were significantly higher than those of the control group (35.0±2.7)mmHg (t=-90.01,P=0.000),(30.0±3.1)mmHg (t=-81.90,P=0.000),(26.0±3.0)mmHg (t=-88.89,P=0.000). Thus,there was a positive correlation between NT-proBNP and IL-6 levels (r=0.876,P=0.000) and a positive correlation between NT-proBNP and PAP (r=0.916,P=0.000) in preterm infants with RDS.Conclusion Monitoring serum NT-proBN contributes to early diagnosis and disease severity assessment in preterm infants with RDS.
Biomarkers ; Early Diagnosis ; Humans ; Infant ; Infant, Newborn ; Infant, Premature ; Interleukin-6 ; Natriuretic Peptide, Brain ; Peptide Fragments ; Respiratory Distress Syndrome, Adult

Objective To discuss the application value of diatom examination in lung tissue for the forensic diagnosis of drowning. Methods The experimental animals were divided randomly into drowning, postmortem submergence and dying on land group. Diatoms in lung tissue and drowning fluid were analyzed quantitatively by microwave digestion-vacuum filtration-automated scanning electron microscopy diatom examination method. The ratios of content of diatoms in lung tissue and drowning fluid （C_L/C_D ratio） were recorded. Results The C_L/C_D ratios of experimental rabbits in the drowning group （5.82±3.50） were much higher than that of postmortem submergence group （0.47±0.35）; the C_L/C_D ratios of different parts of the lung lobes of experimental pigs in the drowning group were higher than that of postmortem submergence group （P<0.05）; in seawater, brackish water, river fresh water and lake fresh water, the C_L/C_D ratios of experimental pigs in the drowning group were higher than that of postmortem submergence group （P<0.05）. In animal experiments, all the cases with C_L/C_D ratio >1.6 were from drowning group. Conclusion C_L/C_D ratio is an indicator with good application prospects in the diagnosis of drowning.
Animals ; Autopsy ; Diatoms/cytology* ; Drowning/diagnosis* ; Lung ; Microscopy, Electron, Scanning ; Rabbits ; Random Allocation ; Swine