Objective:To analyze the infection status and epidemiological characteristics of respiratory pathogens in children with acute respiratory infections (ARI) at Xuzhou Children′s Hospital Affiliated to Xuzhou Medical University from 2023 to 2024.Methods:This study enrolled the patients (aged 0-17 years) who visited the outpatient or emergency department or were hospitalized at Xuzhou Children′s Hospital Affiliated to Xuzhou Medical University due to ARI from March 2023 to March 2024. Throat swab specimens of the patients were collected, and fluorescence quantitative PCR was used to detect influenza A virus (FluA), influenza B virus (FluB), respiratory syncytial virus (RSV), human rhinovirus (HRV), human adenovirus (HAdV), and Myocoplasima pneumonia ( Mp). These patients were divided into five groups by gender: <1, 1-2, 3-5, 6-11, 12-17 years. Chi-square test was used to perform statistical analysis on the detection rates of respiratory pathogens among patients of different genders and ages, and across distinct seasons. Results:A total of 46 379 children were enrolled and among them, 27 418 children tested positive for respiratory pathogens, with a positive rate of 59.12%. Among the positive cases, 5 177 (18.88%) were infected with more than one respiratory pathogen, with the co-infection of Mp and HRV being the most common type, followed by Mp and HAdV co-infection. The pathogens, ranked from the highest to the lowest detection rates, were Mp (20.74%, 9 620/46 379), RSV (12.76%, 5 920/46 379), HAdV (11.64%, 5 399/46 379), HRV (11.24%, 5 213/46 379), FluB (8.23%, 3 815/46 379), and FluA (6.80%, 3 154/46 379). There were statistically significant differences in the detection rates of RSV, HRV and Mp among children of different genders (χ 2=11.85, 15.23, 16.36; all P<0.001). The differences in the detection rates of the six pathogens among different age groups were statistically significant (all P<0.001), and the detection rates of FluA, FluB, HRV, HAdV and Mp in children aged 0-5 years showed an upward trend with age (all P<0.001). The highest detection rates of FluA, FluB, HRV and HAdV were in the 3-5 years group, while the highest detection rate of Mp was in the 6-11 years group, which was 40.15% (4 615/11 495). The detection rate of RSV showed a decreasing trend with age ( P<0.001), with the highest detection rate observed in the <1 year group (25.02%, 2 208/8 826). There were statistically significant differences in the detection rates of the six pathogens in different seasons (all P<0.001). Conclusions:The overall detection rate of respiratory pathogens in children with ARI in Xuzhou from 2023 to 2024 is high. Single-pathogen infection is the predominant pattern, and the most prevalent pathogen is Mp. There are gender differences in the detection rates of RSV, HRV, and Mp. The detection rate of RSV decreases with age, while the detection rates of FluA, FluB, HRV, HAdV, and Mp increase with age among children aged 0-5 years. The prevalence of FluA, FluB, RSV, HRV, HAdV, and Mp all exhibit seasonal patterns.

Objective:To analyze the infection status and epidemiological characteristics of respiratory pathogens in children with acute respiratory infections (ARI) at Xuzhou Children′s Hospital Affiliated to Xuzhou Medical University from 2023 to 2024.Methods:This study enrolled the patients (aged 0-17 years) who visited the outpatient or emergency department or were hospitalized at Xuzhou Children′s Hospital Affiliated to Xuzhou Medical University due to ARI from March 2023 to March 2024. Throat swab specimens of the patients were collected, and fluorescence quantitative PCR was used to detect influenza A virus (FluA), influenza B virus (FluB), respiratory syncytial virus (RSV), human rhinovirus (HRV), human adenovirus (HAdV), and Myocoplasima pneumonia ( Mp). These patients were divided into five groups by gender: <1, 1-2, 3-5, 6-11, 12-17 years. Chi-square test was used to perform statistical analysis on the detection rates of respiratory pathogens among patients of different genders and ages, and across distinct seasons. Results:A total of 46 379 children were enrolled and among them, 27 418 children tested positive for respiratory pathogens, with a positive rate of 59.12%. Among the positive cases, 5 177 (18.88%) were infected with more than one respiratory pathogen, with the co-infection of Mp and HRV being the most common type, followed by Mp and HAdV co-infection. The pathogens, ranked from the highest to the lowest detection rates, were Mp (20.74%, 9 620/46 379), RSV (12.76%, 5 920/46 379), HAdV (11.64%, 5 399/46 379), HRV (11.24%, 5 213/46 379), FluB (8.23%, 3 815/46 379), and FluA (6.80%, 3 154/46 379). There were statistically significant differences in the detection rates of RSV, HRV and Mp among children of different genders (χ 2=11.85, 15.23, 16.36; all P<0.001). The differences in the detection rates of the six pathogens among different age groups were statistically significant (all P<0.001), and the detection rates of FluA, FluB, HRV, HAdV and Mp in children aged 0-5 years showed an upward trend with age (all P<0.001). The highest detection rates of FluA, FluB, HRV and HAdV were in the 3-5 years group, while the highest detection rate of Mp was in the 6-11 years group, which was 40.15% (4 615/11 495). The detection rate of RSV showed a decreasing trend with age ( P<0.001), with the highest detection rate observed in the <1 year group (25.02%, 2 208/8 826). There were statistically significant differences in the detection rates of the six pathogens in different seasons (all P<0.001). Conclusions:The overall detection rate of respiratory pathogens in children with ARI in Xuzhou from 2023 to 2024 is high. Single-pathogen infection is the predominant pattern, and the most prevalent pathogen is Mp. There are gender differences in the detection rates of RSV, HRV, and Mp. The detection rate of RSV decreases with age, while the detection rates of FluA, FluB, HRV, HAdV, and Mp increase with age among children aged 0-5 years. The prevalence of FluA, FluB, RSV, HRV, HAdV, and Mp all exhibit seasonal patterns.

Objective:To investigate the clinical significance and possible mechanisms of elevated homocysteine(Hcy) levels in peripheral blood of children with sepsis.Methods:The clinical data of 51 children with sepsis (sepsis group) admitted to PICU at Xuzhou Children′s Hospital from January 2019 to December 2019 were analyzed, and the levels of Hcy in plasma were compared with 50 non-septic children (common infection group) and 50 healthy children (healthy control group) during the same period.The possible mechanism of metabolic disorders about Hcy was analyzed by detecting the levels of the key rate-limiting enzymes cystathionine-β-synthase(CBS) and cystathionine-γ-lyase(CSE), which were in the downstream of metabolism in septic mouse model induced by lipopolysaccharide.Results:The level of Hcy in plasma was (12.62±5.46)μmol/L in sepsis group, which was significantly higher than those in common infection group[(9.42±2.28) μmol/L] and healthy control group[(8.14±1.60) μmol/L]( P<0.05). The level of Hcy in plasma of 12 children with acute kidney injury in sepsis group was significantly higher than that of 39 children without acute kidney injury in sepsis group[(16.48±5.87)μmol/L vs.(11.62±4.74) μmol/L, P<0.05]. The level of Hcy in plasma of six children with acute liver failure in sepsis group was significant higher than that of 45 children without acute liver failure in sepsis group[(18.35±7.10) μmol/L vs.(11.84±4.78) μmol/L, P<0.05]. The level of Hcy in serum significantly increased in septic mouse models ( P<0.01). The transcription and protein expression levels of key rate-limiting Hcy transcription enzymes CBS and CSE in liver and kidney tissues of septic mouse were significantly down-regulated ( P<0.05). Conclusion:The level of Hcy in peripheral blood of children with sepsis increases, which is more obviously in children with acute kidney injury or acute liver injury.When patients developed sepsis, the expression of CBS and CSE will be restrained, leading to disorders related to transsulfuration metabolism and elevated level of Hcy in peripheral blood.

Objective To investigate the effect of short-term complications after D2 radical gastrectomy on long-term survival rate of gastric cancer patients. Methods A retrospective case-control study was conducted on 421 patients with gastric cancer who underwent D2 radical gastrectomy. According to the short-term postoperative complications, they were divided into experimental group (complication group, n=76) and control group (without complication group, n=345). In order to reduce the selection bias, the long-term survival rate of the two groups was tested by Kaplan-Meier survival analysis method after balancing the variables by propensity score matching (PSM). Log rank method was used for univariate analysis and Cox multivariate analysis was used for prognostic factors. Results There was no significant difference in long-term survival rate between the experimental group and the control group (P > 0.05). Histological type, lymph node metastasis rate and pTNM stage were independent risk factors for long-term survival. Conclusion The short-term complications after D2 radical gastrectomy have no significant impact on the long-term survival rate of gastric cancer patients, but it has certain clinical significance to actively prevent and control complications.

Objective: To investigate the role of hexokinase Ⅱ in the changes of autophagic flow in cardiomyocytes of mice with ischemia-hypoxia in vitro. Methods: The hearts of totally six male and female C57BL/6 mice aged from 1 to 2 days were isolated to culture primary cardiomyocytes which were used for the following experiments. (1) The cells were divided into 6 groups according to the random number table (the same grouping method below), i. e., normal control 3, 6, and 9 h groups and ischemia-hypoxia 3, 6, and 9 h groups, with 4 wells in each group. After being regularly cultured for 48 h with Dulbecco′s modified Eagle medium/nutrient mixture F12 (DMEM/F12) medium (the same regular culture condition below), the cells in normal control 3, 6, and 9 h groups were cultured with replaced fresh DMEM/F12 medium for 3, 6, and 9 h, respectively, and the cells in ischemia-hypoxia 3, 6, and 9 h groups were cultured with replaced sugar-free serum-free medium in the low-oxygen incubator with a volume fraction of 1% oxygen and a volume fraction of 5% carbon dioxide at 37 ℃ (the same hypoxic culture condition below) for 3, 6, and 9 h, respectively. Cell viability was measured by the cell counting kit 8 (CCK-8) method. (2) The cells were grouped and treated the same as those in experiment (1), with 1 well in each group. Western blotting was used to detect the protein expressions of microtubule-associated protein 1 light chain 3 Ⅰ (LC3Ⅰ), LC3Ⅱ, p62, and hexokinase Ⅱ. (3) The cells were divided into normal control group, simple ischemia-hypoxia 9 h group, and ischemia-hypoxia 9 h+ 2-deoxyglucose (2-DG) group, with 4 wells in each group. After a regular culture for 48 h, the cells in normal control group were cultured with replaced fresh DMEM/F12 medium for 9 h; the cells in simple ischemia-hypoxia 9 h group were replaced with sugar-free serum-free medium, and the cells in ischemia-hypoxia 9 h+ 2-DG group were replaced with sugar-free serum-free medium in which 2-DG was dissolved in a concentration of 10 mmol/L (20 μmol), and then they were cultured with hypoxia for 9 h. Cell viability was measured by CCK-8 method. (4) The cells were grouped and treated the same as those in experiment (3), with 1 well in each group. Western blotting was used to detect the protein expressions of LC3Ⅰ, LC3Ⅱ, and p62. (5) The cells were grouped and treated the same as those in experiment (3), with 2 wells in each group. Transmission electron microscope was used to observe autophagosomes/autolysosomes in cardiomyocytes. (6) The cells were divided into normal control group, simple ischemia-hypoxia 9 h group, ischemia-hypoxia 9 h+ hexosinase Ⅱ small interfering RNA1 (HK-ⅡsiRNA1) group, and ischemia-hypoxia 9 h+ HK-ⅡsiRNA2 group, with 4 wells in each group. The cells in normal control group and simple ischemia-hypoxia 9 h group were regularly cultured for 48 h, and the cells in ischemia-hypoxia 9 h+ HK-ⅡsiRNA1 group and ischemia-hypoxia 9 h+ HK-ⅡsiRNA2 group were respectively transfected with 200 nmol/L HK-ⅡsiRNA1 and HK-ⅡsiRNA2 and then also cultured for 48 h. The cells in normal control group were cultured with replaced fresh DMEM/F12 medium for 9 h, and the cells in simple ischemia-hypoxia 9 h group, ischemia-hypoxia 9 h+ HK-ⅡsiRNA1 group, and ischemia-hypoxia 9 h+ HK-ⅡsiRNA2 group were cultured with replaced sugar-free serum-free medium and hypoxia for 9 h. Cell viability was measured by CCK-8 method. (7) The cells were grouped and treated the same as those in experiment (6), with 1 well in each group. Western blotting was used to detect the protein expressions of LC3Ⅰ, LC3Ⅱ, p62, and hexokinase Ⅱ. Except for experiment (5), each experiment was repeated 3 times. Data were processed with one-way analysis of variance and lest significant difference t test, and Bonferroni correction. Results: (1) The viabilities of cardiomyocytes in ischemia-hypoxia 3, 6, and 9 h groups were 0.450±0.022, 0.385±0.010, and 0.335±0.015, respectively, which were significantly lower than 0.662±0.026, 0.656±0.028, and 0.661±0.021 of the corresponding normal control 3, 6, and 9 h groups, respectively (t=6.21, 9.12, 12.48, P<0.01). (2) Compared with those of corresponding normal control 3, 6, and 9 h groups, the LC3Ⅱ/Ⅰ ratio and protein expressions of p62 and hexokinase Ⅱ in cardiomyocytes of ischemia-hypoxia 3, 6, and 9 h groups were significantly increased (t_{3 h}=16.15, 10.99, 5.30, t_{6 h}=6.79, 10.42, 9.42, t_{9 h}=15.76, 16.51, 7.20, P<0.05 or P<0.01). (3) The viability of cardiomyocytes in simple ischemia-hypoxia 9 h group was 0.353±0.022, which was significantly lower than 0.673±0.027 of normal control group (t=9.29, P<0.01). The viability of cardiomyocytes in ischemia-hypoxia 9 h+ 2-DG group was 0.472±0.025, which was significantly higher than that of simple ischemia-hypoxia 9 h group (t=3.60, P<0.05). (4) Compared with those of normal control group, the LC3Ⅱ/Ⅰ ratio and protein expression of p62 in cardiomyocytes of simple ischemia-hypoxia 9 h group were significantly increased (t=9.45, 8.40, P<0.01). Compared with those of simple ischemia-hypoxia 9 h group, the LC3Ⅱ/Ⅰratio and protein expression of p62 in cardiomyocytes of ischemia-hypoxia 9 h+ 2-DG group were significantly decreased (t=4.39, 4.74, P<0.05). (5) In cardiomyocytes of normal control group, only single autophagosome/autolysosome with bilayer membrane structure was observed. Compared with that of normal control group, the number of autophagosome/autolysosome with bilayer membrane structure in cardiomyocytes of simple ischemia-hypoxia 9 h group was increased significantly. Compared with that of simple ischemia-hypoxia 9 h group, the number of autophagosome/autolysosome with bilayer membrane structure in cardiomyocytes of ischemia-hypoxia 9 h+ 2-DG group was significantly decreased. (6) The viability of cardiomyocytes in simple ischemia-hypoxia 9 h group was 0.358±0.023, which was significantly lower than 0.673±0.026 in normal control group (t=9.12, P<0.01). The viabilities of cardiomyocytes in ischemia-hypoxia 9 h+ HK-ⅡsiRNA1 group and ischemia-hypoxia 9 h+ HK-ⅡsiRNA2 group were 0.487±0.027 and 0.493±0.022, respectively, which were significantly higher than the viability in simple ischemia-hypoxia 9 h group (t=3.63, 4.28, P<0.05). (7) Compared with those of normal control group, the LC3Ⅱ/Ⅰratio and protein expressions of p62 and hexokinase Ⅱ in cardiomyocytes of simple ischemia-hypoxia 9 h group were significantly increased (t=6.08, 6.31, 4.83, P<0.05 or P<0.01). Compared with those of simple ischemia-hypoxia 9 h group, the LC3Ⅱ/Ⅰ ratio and protein expressions of p62 and hexokinase Ⅱ in cardiomyocytes of ischemia-hypoxia 9 h+ HK-ⅡsiRNA1 group and ischemia-hypoxia 9 h+ HK-ⅡsiRNA2 group were significantly decreased (t=5.10, 7.76, 15.33, 4.17, 8.42, 12.11, P<0.05 or P<0.01). Conclusions Ischemia-hypoxia upregulates the expression level of hexokinase Ⅱ protein in mouse cardiomyocytes cultured in vitro, which decreases the viability of cardiomyocytes by impairing autophagic flow. To inhibit the activity of hexokinase Ⅱ or its expression can alleviate the ischemia-hypoxia damage of cardiomyocytes.

Objective: To investigate the effect of human antigen R on lysosomal acidification during autophagy in mouse cardiomyocytes cultured in vitro. Methods: The hearts of 20 C57BL/6 mice aged 1-2 days no matter male or female were isolated to culture primary cardiomyocytes which were used in the following experiments. (1) The cells were divided into 5 groups according to the random number table (the same grouping method below), i. e., normal control group and sugar-free serum-free 0.5, 1.0, 3.0, and 6.0 h groups. The cells in normal control group were routinely cultured for 54.0 h with Dulbecco′s modified Eagle medium/nutrient mixture F12 (DMEM/F12) medium (the same regular culture condition below), and the cells in sugar-free serum-free 0.5, 1.0, 3.0, and 6.0 h groups were firstly regularly cultured for 53.5, 53.0, 51.0, 48.0 h and then cultured with replaced sugar-free serum-free medium for 0.5, 1.0, 3.0, and 6.0 h, respectively. The protein expressions of microtubule-associated protein 1 light chain 3 Ⅱ (LC3Ⅱ), autophagy-related protein 5, and adenosine triphosphatase V1 region E1 subunit (ATP6V1E1) were detected by Western blotting. (2) The cells were divided into normal control group and sugar-free serum-free 3.0 h group. The cells in corresponding groups were treated the same as those in experiment (1), and the cell lysosomal acidification level was observed and detected under a laser scanning confocal microscope. (3) Two batches of cells were grouped and treated the same as those in experiment (1). The protein expression of human antigen R in the whole protein of cells of one batch and its protein expression in the cytoplasm and nucleus protein of cells of the other batch were detected by Western blotting. (4) The cells were divided into normal control group, simple control small interfering RNA (siRNA) group, simple human antigen R-siRNA1 (HuR-siRNA1) group, simple HuR-siRNA2 group, sugar-free serum-free 3.0 h group, sugar-free serum-free+ control siRNA group, sugar-free serum-free+ HuR-siRNA1 group, and sugar-free serum-free+ HuR-siRNA2 group. After 48 hours of regular culture, the cells in simple control siRNA group and sugar-free serum-free+ control siRNA group were transfected with negative control siRNA for 6 h, the cells in simple HuR-siRNA1 group and sugar-free serum-free+ HuR-siRNA1 group were transfected with HuR-siRNA1 for 6 h, and the cells in simple HuR-siRNA2 group and sugar-free serum-free+ HuR-siRNA2 group were transfected with HuR-siRNA2 for 6 h. Hereafter, the cells in these 8 groups were continuously cultured for 48 h with regular conditon, and then the cells in normal control group and each simple siRNA-treated group were replaced with DMEM/F12 medium, the cells in the other groups were replaced with sugar-free serum-free medium, and they were cultured for 3 h. The protein expression of human antigen R in the whole protein of cells was detected by Western blotting. (5) Two batches of cells were divided into sugar-free serum-free+ control siRNA group and sugar-free serum-free+ HuR-siRNA1 group, and the cells in corresponding groups were treated the same as those in experiment (4). The distribution and expression of human antigen R in the cells of one batch were observed and detected by immunofluorescence method, and the lysosomal acidification level in the cells of the other batch was observed and detected under a laser scanning confocal microscope. (6) Three batches of cells were divided into sugar-free serum-free 3.0 h group, sugar-free serum-free+ control siRNA group, sugar-free serum-free+ HuR-siRNA1 group, and sugar-free serum-free+ HuR-siRNA2 group, and the cells in corresponding groups were treated the same as those in experiment (4). The protein expressions of cathepsin D in the whole protein of cells of one batch, human antigen R in the cytoplasm protein of cells of one batch, and ATP6V1E1 in the whole protein of cells of the other batch were detected by Western blotting. (7) The cells were divided into normal control group, sugar-free serum-free 3.0 h group, sugar-free serum-free+ control siRNA group, and sugar-free serum-free+ HuR-siRNA1 group, and the cells in corresponding groups were treated the same as those in experiment (4). The mRNA expression of ATP6V1E1 in cells was detected by real-time fluorescent quantitative reverse transcription polymerase chain reaction. The sample number of each experiment was 3. Data were processed with independent data t test, one-way analysis of variance, least significant difference t test, and Bonferroni correction. Results: (1) Compared with those of normal control group, the protein expressions of LC3Ⅱ and ATP6V1E1 in the whole protein of cells of sugar-free serum-free 1.0, 3.0, and 6.0 h groups were significantly increased (t=12.16, 4.05, 4.82, 11.64, 3.29, 8.37, P<0.05 or P<0.01). Compared with that of normal control group, the protein expression of autophagy-related protein 5 in the whole protein of cells of sugar-free serum-free 0.5, 1.0, 3.0, and 6.0 h groups was significantly increased (t=6.88, 10.56, 5.76, 9.91, P<0.05 or P<0.01). (2) Compared with 1.03±0.08 of normal control group, the lysosomal acidification level in the cells of sugar-free serum-free 3.0 group (2.92±0.30) was significantly increased (t=6.01, P<0.01). (3) There was no statistically significant difference in the overall comparison of protein expression of human antigen R in the whole protein of cells among the 5 groups (F=1.09, P>0.05). Compared with that of normal control group, the protein expression of human antigen R in the cytoplasm protein of cells was significantly increased in sugar-free serum-free 1.0, 3.0, and 6.0 h groups (t=43.05, 11.07, 5.39, P<0.05 or P<0.01), while the protein expression of human antigen R in the nucleus protein of cells was significantly decreased in sugar-free serum-free 3.0 and 6.0 h groups (t=11.18, 12.71, P<0.01). (4) Compared with that of simple control siRNA group, the protein expression of human antigen R in the whole protein of cells of simple HuR-siRNA1 group and simple HuR-siRNA2 group was significantly decreased (t=4.82, 4.44, P<0.05). Compared with that of sugar-free serum-free+ control siRNA group, the protein expression of human antigen R in the whole protein of cells of sugar-free serum-free+ HuR-siRNA1 group and sugar-free serum-free+ HuR-siRNA2 group was significantly decreased (t=4.39, 6.27, P<0.05). (5) Compared with those of sugar-free serum-free+ control siRNA group, the distribution of human antigen R in the cytoplasm of cells and its expression level were significantly decreased in sugar-free serum-free+ HuR-siRNA1 group (t=10.13, P<0.01). Compared with 1.00±0.06 of sugar-free serum-free+ control siRNA group, the lysosomal acidification level (0.73±0.06) in the cells of sugar-free serum-free+ HuR-siRNA1 group was significantly decreased (t=3.28, P<0.01). (6) Compared with those of sugar-free serum-free+ control siRNA group, the protein expressions of cathepsin D in the whole protein of cells, human antigen R in the cytoplasm protein of cells, and ATP6V1E1 in the whole protein of cells were significantly decreased in sugar-free serum-free+ HuR-siRNA1 group and sugar-free serum-free+ HuR-siRNA2 group (t=4.16, 3.99, 4.81, 5.07, 11.68, 12.97, P<0.05 or P<0.01). (7) Compared with that of normal control group, the mRNA expression of ATP6V1E1 in the cells of sugar-free serum-free 3.0 h group was significantly increased (t=5.51, P<0.05). Compared with that of sugar-free serum-free+ control siRNA group, the mRNA expression of ATP6V1E1 in the cells of sugar-free serum-free+ HuR-siRNA1 group was significantly decreased (t=5.97, P<0.05). Conclusions After sugar-free serum-free treatment in vitro, the autophagy in mouse primary cardiomyocytes is activated, the lysosomal acidification is enhanced, and the expression of human antigen R in cytoplasm is increased. Human antigen R function is activated and involved in maintaining lysosomal acidification during autophagy in mouse cardiomyocytes.

Objective: To investigate the application value of cerebrospinal fluid circulating cell-free DNA (cfDNA) in the diagnosis and treatment of leptomeningeal metastases in non-small-cell lung cancer (NSCLC). Methods: Twenty-five patients with leptomeningeal metastases of NCSLC from Fudan University Huashan Hospital North during the period from September 2017 to November 2018 were enrolled. All 25 patients were confirmed leptomeningeal metastases by cerebrospinal fluid cytology and immunocytochemical staining of cytokeratin(CK7), carcinoembryonic antigen(CEA), thyroid transcription factor-1(TTF-1) and Ki67. The cerebrospinal fluid cfDNA was extracted and genetic variation of 12 genes including epidermal growth factor receptor(EGFR), TP53 and anaplastic lymphoma kinase(ALK) was detected by next-generation sequencing [PlasAim TM gene non-invasive detection of lung cancer (12 gene) kit, Singlera Genomics].The application value of cerebrospinal fluid cfDNA in the diagnosis and treatment of leptomeningeal metastases of NSCLC was analyzed with the cfDNA mutation data and the clinical follow-ups. Results: Morphologically typical lung cancer tumor cells with tumor immunochemistry markerCK, CK7 and CEA were found in the cerebrospinal fluid of all 25 patients. Next generation sequencing of cerebrospinal fluid showed that 96% (24/25) patients had at least one single nucleotide variation (SNV) or copy number variation (CNV). The EGFR and TP53 mutations were identified in 80% (20/25) and 48%(12/25) of the patients, respectively. In addition, patients with bone metastases had a higher rate of EGFR mutations than those without bone metastases (100% vs 64%, P<0.05). Changes in the mutant allele frequency of EGFR and TP53 in cerebrospinal fluid were consistent with patients′ disease progression parameters including neurological symptoms, imaging, and tumor biomarkers. The results indicate that genetic alteration of EGFR in cerebrospinal fluid cfDNA is an actionable biomarker for targeted therapy of leptomeningeal metastases of lung cancer. Conclusion Cerebrospinal fluid cfDNA accurately reveals the unique genetic background of leptomeningeal metastasis in NSCLC, showing great application value in the diagnosis and treatment of the leptomeningeal metastasis of NSCLC.

BACKGROUND:It is the key point to choose the right size of the prosthesis, and grasp the direction and thickness for osteotomy during total knee arthroplasty. In order to achieve the goal, accurate preoperative planning is very important.
OBJECTIVE:To compare the accuracy of preoperative templating in total knee arthroplasty using conventional two-dimensional (2D) and computed tomography (CT)-based three-dimensional (3D) procedures (templating on 3D image&surgical rehearsing on rapid prototype technology-models), and to confirm the necessity of 3D evaluation for preoperative planning.
METHODS:A total of 25 patients undergoing primary total knee arthroplasty were randomly selected, including 10 males and 15 females, at the age of 58 and 79 years old. 2D and 3D images were col ected from al patients. Preoperative templating was performed for each total knee arthroplasty using both conventional 2D radiographs and a CT-based 3D image model. Accuracies with regard to the predicted and actual implant sizes were determined for each procedure.
RESULTS AND CONCLUSION:The 3D procedure was found to be more accurate in predicting implant size of 80%femoral and 72%tibial components than those of the 2D procedure (4%femoral and 12%tibial components). Significant differences in the consistent rate of femoral and tibial prosthesis models were detected significantly (P<0.05). Kappa coefficient statistics demonstrated that goodness of fit of prosthesis model was good in 3D preoperative templating. Results confirmed that the superiority of 3D preoperative templating over 2D conventional evaluation is in predicting implant size, and provides more comprehensive information on skeletal anatomy.

BACKGROUND:Posterior stabilized femoral knee prosthesis needs additional condyle osteotomy to accommodate the tibial post and femur fossa structures. Intercondylar fossa on both sides connected at the femoral body with concentrated stress is a place easily affecting fractures. Differences in bone mass between different models of different brands did not have specific data, which was not convenient to select prosthesis for clinicians.
OBJECTIVE: To compare the difference of intercondylar osteotomy data among clinical commonly used posterior stabilized knee prostheses (six imported and domestic brands), and to provide basis for the selection and application of the prostheses.
METHODS:The current commonly used posterior stabilized knee prostheses (six imported and domestic brands) were used, including Zimmer NexGen LPS, Stryker Scrorpio NRG Knee-Flexed, Depuy PFC Sigma, Smith & nephew Genesis-2 PS, United-U1 and Wego GKPS. According to the osteotomy template, the osteotomy-surfaces consisting of femoral condyle starting section and cross section, distal section of femoral condyle, and back-oblique section were identified. The corresponding femoral prosthesis diameter lines included condylar ambilateral and anteroposterior diameters, width and depth of femoral intercondylar fossa. The above data were compared and measured.
RESULTS AND CONCLUSION:The six kinds of knee femoral prostheses were different in ratio of ambilateral diameter and anteroposterior diameter, bone resection of intercondylar fossa, and geometry. Imported prostheses carry shorter diameters in femoral starting and cross sections, so it can catch more posterior condylar osteotomy. With increasing prosthesis sizes, the ratio of bone loss causing by width of intercondylar osteotomy is decreased among six brands. In al sizes, Stryker Scrorpio NRG Knee-Flexed catches shorter width of intercondylar osteotomy. Knee prosthesis osteotomy among six brands is different. The result of this study is not sufficient to evaluate the pros and cons between different prostheses, but as reserving bone is concerned, the design of less intercondylar osteoomy catches more advantages.

Objective To investigate the perioperative risk of deep vein thrombosis (DVT) in patients with hepatic cirrhosis that underwent total hip arthroplasty (THA), and to evaluate the safety and feasibility of individualized anti-coagulation treatment.Methods There were 25 patients complicating hepatic cirrhosis that underwent THA (from Jan.to Dec.2014), including 17 males and 8 females, aged 57.9t9.2 years.The primary causes of THA were avascular necrosis of the femoral head (eighteen cases) and osteoarthritis of the hip (seven cases).Low molecular weight heparin (LMWH) was applied for anti-coagulation treatment.Parameters of hepatic function and coagulation function of THA cases (randomized thirty cases, from Jan.2008 to Dec.2008) without hepatic cirrhosis were used as reference for monitoring.For the cases of massive blood loss or upper gastrointestinal hemorrhage, a LMWH administration pause and an administration of fresh frozen plasma and clotting factors were performed in order to maintain a hemorrage/coagulation balance.The clinical outcome of the hip joint was evaluated and complications were treated.A subsequent follow-up was also carried out after perioperative period.Results All cases received successful surgeries and followed up.The follow-up duration was 34± 15.7 months.The preoperative Harris hip score was 32.4± 10.2 points, while the most recent follow-up score was 82.9±6.1 points, which was statistically significant.Dislocation, periprosthetic fracture and periprosthetic infection were absent.All cases received individualized anti-coagulation treatments during peripoerative period.A hemorrage/coagulation balance was achieved.The dynamic parameter curves did not present excessive deviation from reference.One case encountered intermuscular hematoma of the lower limbs 48 hours postoperatively, which was solved by a LMWH pause and administration of fresh frozen plasma and clotting factors.One case suffered upper gastrointestinal hemorrhage five days postoperatively, which was controlled by a LMWH pause and the administration of somatostatin and proton pump inhibitor.Jaundic got worse in one case three days postoperatively but got relieved after treatment.Overt blood loss was 686t141.8 ml.Perioperative death, hepatic failure, hepatic encephalopath, hepatorenal syndrome were absent.No DVT was observed.Conclusion There are risks of DVT in patients of hepatic cirrhosis.Individualized anti-coagulation treatment is needed during perioperative period of THA.