This study aimed to identify,characterize,and functionally analyze the CYP450 gene fam-ily within Hyalomma asiaticum unfed nymphs(HasiUN)under regular physiological conditions,laying the groundwork for further study into the biosynthesis and metabolism of exogenous sub-stances associated with the CYP450 gene in HasiUN.Through our methods,we identified and characterized HasiUNCYP450 genes.We then annotated them using the gene ontology(GO)and kyoto encyclopedia of genes and genomes(KEGG).This was achieved with the help of bioinforma-tics,based on the transcriptome of 2-week-old HasiUN.Further on,we conducted a preliminary a-nalysis of tissue expression,distribution,and potential function with qRT-PCR and network inter-action predictions.Our analyses showed that 87 HasiUN CYP450 genes,ranging in size from 1089 to 1692 bp,encoded predominantly basic,hydrophilic,and unstable protein sequences.These se-quences had nearly identical numbers of transmembrane regions,both with and without a signal peptide.Phylogenetic and domain analyses indicated that HasiUN CYP450,having a complete CYP450 domain,were distributed among various clans,with most members found in clan 3.CYP20 clans,present in 13 species from 2 families and 7 genera,required either one-host or three-hosts to undergo their full life cycle.MEME analysis results hint at different clans having variations in the amount and combination of motifs.Expression level analysis showed that HasiUN CYP450 was expressed at different levels,with the highest distribution of normally expressed genes in clan 3 and low expression of all mito clan genes.Functional annotation revealed HasiUN CYP450 key as-sociation with biological processes,metabolism,and organismal systems according to the GO and KEGG annotations.Distribution results showed that HasiUN CYP20a1 was expressed in specific tissues,including salivary glands and ovaries.Functional analysis points to CYP20a1's role in UV-induced toxic metabolism,endocrine disruptor metabolism,and hazardous substance metabolism in the nervous system.Our research presents the first analysis of HasiUN CYP450 gene's biological characteristics and expression pattern under normal physiological conditions.Additionally,investi-gations into the distribution and function of a new category,HasiUN CYP20a1,were embarked upon.These findings establish a theoretical foundation for continued functional study of HasiUN CYP45 gene.

This study aimed to identify,characterize,and functionally analyze the CYP450 gene fam-ily within Hyalomma asiaticum unfed nymphs(HasiUN)under regular physiological conditions,laying the groundwork for further study into the biosynthesis and metabolism of exogenous sub-stances associated with the CYP450 gene in HasiUN.Through our methods,we identified and characterized HasiUNCYP450 genes.We then annotated them using the gene ontology(GO)and kyoto encyclopedia of genes and genomes(KEGG).This was achieved with the help of bioinforma-tics,based on the transcriptome of 2-week-old HasiUN.Further on,we conducted a preliminary a-nalysis of tissue expression,distribution,and potential function with qRT-PCR and network inter-action predictions.Our analyses showed that 87 HasiUN CYP450 genes,ranging in size from 1089 to 1692 bp,encoded predominantly basic,hydrophilic,and unstable protein sequences.These se-quences had nearly identical numbers of transmembrane regions,both with and without a signal peptide.Phylogenetic and domain analyses indicated that HasiUN CYP450,having a complete CYP450 domain,were distributed among various clans,with most members found in clan 3.CYP20 clans,present in 13 species from 2 families and 7 genera,required either one-host or three-hosts to undergo their full life cycle.MEME analysis results hint at different clans having variations in the amount and combination of motifs.Expression level analysis showed that HasiUN CYP450 was expressed at different levels,with the highest distribution of normally expressed genes in clan 3 and low expression of all mito clan genes.Functional annotation revealed HasiUN CYP450 key as-sociation with biological processes,metabolism,and organismal systems according to the GO and KEGG annotations.Distribution results showed that HasiUN CYP20a1 was expressed in specific tissues,including salivary glands and ovaries.Functional analysis points to CYP20a1's role in UV-induced toxic metabolism,endocrine disruptor metabolism,and hazardous substance metabolism in the nervous system.Our research presents the first analysis of HasiUN CYP450 gene's biological characteristics and expression pattern under normal physiological conditions.Additionally,investi-gations into the distribution and function of a new category,HasiUN CYP20a1,were embarked upon.These findings establish a theoretical foundation for continued functional study of HasiUN CYP45 gene.

Objective To explore the imaging features, clinical outcome, and prognosis of indeterminate pulmonary nodules (IPN) in patients with high-grade soft tissue sarcoma. Methods A retrospective study of 82 patients with high-grade soft tissue sarcoma who have IPNs. The clinical characteristics, imaging features of IPN, and survival of patients were analyzed with statistical software. Results The IPN size of 82 patients was 6.453±0.864 mm. IPN diameter, shape, density, and nodule discovery interval may be CT imaging features related to malignancy tendency. Age (HR=1.047, 95%CI: 1.007-1.088) and interval between each nodule discovery (HR=3.194, 95%CI: 1.052-9.694) are independent factors that affect the survival of patients with malignant IPN. Conclusion The imaging features of chest CT may provide important guidance for determining the nature and survival prognosis of benign and malignant nodules.

OBJECTIVE To investigate the potential therapeutic effect of the sporoderm-removed Ganoderma lucidum spore tablet "Xianzhi No.3" from the perspective of immunomodulation and cardioprotection. METHODS Chemical components of the sporoderm-removed Ganoderma lucidum spore tablet "Xianzhi No.3" were analyzed by UPLC-Q-TOF-MS and colorimetric methods. Examined tablet’s effects on zebrafish models of macrophage reduction, heart failure, H2O2-induced oxidative stress in myocardial and endothelial cells, and a microglial inflammation model induced by lipopolysaccharide. Immune regulation and cardioprotective effects were evaluated through multiple indicators, including macrophage formation and phagocytosis abilities, anti-neuroinflammation ability, cardiac systolic and diastolic functions, and anti-oxidative stress injury ability in myocardial and endothelial cells. RESULTS The sporoderm-removed Ganoderma lucidum spore tablet "Xianzhi No.3" improved macrophage formation and phagocytosis, cardiac systolic and diastolic functions, reduced neuroinflammation, and alleviated oxidative stress in myocardial and endothelial cells, resulting in immunomodulatory and cardioprotective effects. CONCLUSION The sporoderm-removed Ganoderma lucidum spore tablet "Xianzhi No.3" maybe a potential therapeutic agent for regulating the immune system and protecting cardiac function.

To predict the distribution and habitat of Dermacentor nuttallii in Xinjiang and to under-stand its life cycle history.In this study,Maximum Entropy Modeling(MaxEnt)was used to pre-dict the geographical distribution and adaptation area of Dermacentor nuttallii in Xinjiang,and jackknife test and response curve of environmental variables were used to evaluate the environmen-tal factors affecting the distribution of Dermacentor nuttallii.Dermacentor samples were collected randomly based on predicted sites.The species of Dermacentor was identified by the combination of morphological characteristics and molecular biology.The New Zealand white rabbit was the only blood donor,and the life cycle and biological characteristics of the tick were observed and recorded by the method of earmuffs feeding under natural light in the laboratory.The jackknife test and SPSS analysis results showed that the main environmental variables affecting the distribution of suitable areas of Dermacentor nuttallii were average temperature(Bio1),average daily tempera-ture range(Bio2),seasonal temperature change(Bio4),driest month precipitation(Bio14),precip-itation variation coefficient(Bio15).The response curves of major environmental variables showed that Dermacentor nuttallii had the highest presence probability when Bio1 was 15.58 ℃,Bio2 was 6.19 ℃,Bio4 had a coefficient of variation of 1 500,Bio14 had a coefficient of variation of 20 mm,Bio15 had a coefficient of variation of 23.799 and Bio19 was 69 mm.The prediction results showed that the suitable areas of Dermacentor nuttallii in Xinjiang were Tianshan Mountain range in Junggar Basin,Turpan basin in Altai Mountain valley and some areas south of Tianshan Mountain range,accounting for 50.99％of the total area of Xinjiang.Dermacentor were collected from the predicted area according to MaxEnt model and identified as Dermacentor nuttallii by morphologi-cal and molecular biological methods.The full blood stage was 5.31 d,the degeneration stage was 8.19 d,and the degeneration rate reached 95.5％.If the full blood stage was 8.65 d and the degener-ation stage was 12.86 d,the degeneration rate reached 98％.The full blood stage of adult ticks was 6.75 d,the early egg stage of full blood females was 5.86 d,and the spawning stage of full blood fe-males was 12.5 d.The eggs hatched into young ticks after 25.92 d,and the hatching rate reached 90％.Dermacentor nuttallii took 62-107 d to complete a life history,with an average of 86.06 d.The constructed MaxEnt model has high prediction accuracy and accuracy.According to the varia-ble analysis of the main environmental factors,precipitation and temperature are the main environ-mental factors affecting Dermacentor nuttallii.The study of the whole life cycle of Dermacentor nuttallii in Xinjiang provides the basis for establishing the method of artificial breeding of pure species of Dermacentor nuttallii in Xinjiang.

Objective:To explore the incidence, clinical characteristics, imaging features, clinical outcome and prognosis of indeterminate pulmonary nodules (IPN) in patients with osteosarcoma.Methods:A total of 69 patients of osteosarcoma with IPN in lung treated in the Bone tumor Center of Eastern Theater General Hospital from January 2011 to January 2021 were collected retrospectively, there were 47 males and 22 females, with a median age of 19 years old (range 7-60 years old). The clinical characteristics including disease-free interval, the chemotherapy response, with recurrence/non-pulmonary, IPN presence before / during / after chemotherapy and imaging features of IPN including number of IPN, location of IPN, density of IPN, boundary clarity of IPN and outcome. The patients were divided into the metastasis pulmonary nodules group and the benign nodules group according to the final outcome of IPN. Further, χ 2 test was performed for comparison of the clinical and imaging characteristics between the two groups. The survival of patients was counted and the correlation between single factor and survival was compared by Kaplan-Meier test, and multivariate survival analyses were performed using Cox proportional hazards regression models. Results:Sixty-nine cases occurred IPN in 211 patients with osteosarcoma, with an incidence of 32.7%. Of the 69 patients, 45 patients (65.2%) with IPN were diagnosed as metastases, and 24 patients (34.8%) with IPN were diagnosed as benign nodules. Follow-up length ranged from 1 to 124 months, with the median follow up time 43 months. To the end of follow-up, 41 patients (59.4%) remained alive and 28 patients (40.6%) had died. The median survival time was 41.0 (20.0, 65.0) months and the median survival time after diagnosis of IPN was 25.0 (10.0, 43.0) months. There were significant differences in lung nodule density ( P<0.001), boundary ( P=0.002), history of recurrence/extra-pulmonary metastasis ( P=0.023) and chemotherapeutic effect ( P<0.001) between the metastasis pulmonary nodules group and the benign nodules group. Multivariate survival analysis showed that chemotherapeutic effect was an independent factor affecting the overall survival of patients [ HR=0.048, 95% CI (0.01, 0.26)]. Boundary definition [ HR=0.12, 95% CI (0.02, 0.93)] and chemotherapeutic effect [ HR=0.06, 95% CI (0.01, 0.29)] were independent factors influencing survival after diagnosis of IPN. Conclusion:Osteosarcoma patients with IPN have a poor prognosis. The poor effect of chemotherapy is an independent risk factor for the overall survival time of those patients and the survival time after diagnosis of IPN. The boundary definition of IPN is an independent risk factor for the survival time after diagnosis of IPN.

Objective:To analyze the value of peritumoral vascular invasion (PVI) on the prognosis of patients with osteosarcoma.Methods:A total of 232 patients with primary osteosarcoma from 2007 to 2016 were retrospectively analyzed, including 142 males and 90 females. The average age was 17.9±8.2 years (range, 3-39 years). There were 22 positive and 210 negative cases of PVI, 94 deaths and 138 survivals. Univariate survival analysis (Log-rank test and univariate Cox regression) was used to evaluate the effects of age, gender, PVI status, tumor location, surgical method, sensitivity to chemotherapy, and chemotherapy regimen on the prognosis of osteosarcoma. The indicators with statistically significant differences were included in the multivariate Cox regression model to finally determine the risk factors affecting the prognosis of osteosarcoma. The relationship between PVI status and 5-year survival and the incidence of recurrence or metastasis was evaluated using the Kaplan-Meier method.Results:All patients were followed up for 7.6±4.5 years (range, 0.1-15 years). The differences in sensitivity to chemotherapy (χ 2=9.52, P=0.002), choice of chemotherapy regimen (χ 2=8.87, P=0.012), choice of surgical modality (χ 2=13.50, P<0.001), tumor metastasis rate (χ 2=8.51, P=0.004) and mortality rate (χ 2=5.39, P= 0.020) of PVI positive group and PVI negative group had statistically significant differences. Univariate survival analysis was performed on 232 patients with osteosarcoma (gender, age, PVI status, site of tumor development, surgical modality, sensitivity to chemotherapy, and chemotherapy regimen). Indicators with statistically significant differences were included in a multifactorial Cox regression model. The results showed PVI positive [5-year survival rate: HR=2.02, 95% CI (1.61, 2.79), P=0.010; 5-year recurrence or metastasis rate: HR=2.25, 95% CI (1.55, 3.14), P<0.001], surgical procedure as amputation [5-year survival rate: HR=1.22, 95% CI (0.94, 1.78), P=0.037; 5-year recurrence or metastasis rate: HR=1.58, 95% CI (1.11, 2.23), P=0.026] and poor sensitivity to chemotherapy [5-year survival rate: HR=2.71, 95% CI (1.84, 3.98), P=0.001; 5-year recurrence or metastasis rate: HR=2.52, 95% CI (1.88, 3.45), P<0.001] was associated with poor prognosis. Kaplan-Meier curve showed that the 5-year survival rate of PVI positive group was 34%, which was lower than 68% of PVI negative group. The 5-year recurrence or metastasis rate was 72% in the PVI negative group, which was significantly higher than 38% in the PVI negative group ( P<0.05). Conclusion:The 5-year survival rate of PVI positive group was lower than that of PVI negative group, and the 5-year recurrence or metastasis rate was higher than that of PVI negative group. The presence of microvascular angiosarcoma plugs infiltrating the peritumoral tissue in surgical specimens of osteosarcoma after neoadjuvant chemotherapy is a useful indicator to assess the prognosis of patients with osteosarcoma.

OBJECTIVES: To establish a rapid method for the analysis of bucinnazine in blood by UPLC-MS/MS and to apply the method to the practical case. METHODS: After the internal standard was added to blood, the protein was precipitated with 900 μL mixed solution (V_acetonitrile∶V_water=8∶2). After vortex and centrifugation, the protein was measured through 0.22 μm filter membrane. The separation was performed on C₁₈ chromatography column, with acetonitrile and 5 mmol/L ammonium acetate containing 0.1% formic acid aqueous as mobile phase gradient elution at the flow rate of 0.4 mL/min. Multiple reaction monitoring scan was performed in electrospray positive ion mode, quantitative measurement was performed by internal standard method, and methodological verification was carried out. RESULTS: The linear relationship of bucinnazine in blood was good in the range of 0.5-200 μg/L, the correlation coefficient (r) was 0.999 7, the limit of detection was 0.1 μg/L, the limit of quantitation was 0.5 μg/L, and the recovery was 78.3%-83.8% at 1, 10 and 100 μg/L mass concentration levels. The matrix effect was 69.4%-73.8%, the intra-day precision was 1.9%-2.8%, and the inter-day precision was 2.8%-3.2%, the accuracy was 3.1%-3.5%. The stability test results of 1 and 100 μg/L mass concentrations at -25 ℃ showed that the accuracy (bias) of 10 d was less than 4.5%. CONCLUSIONS This method has the advantages of simple pre-treatment process, fast sample processing speed, high sensitivity of instrument analysis, good stability of content determination and reliable identification results, and can meet the needs of case identification.
Tandem Mass Spectrometry/methods* ; Chromatography, Liquid ; Chromatography, High Pressure Liquid/methods* ; Acetonitriles

Objective:To evaluate catheter directed thrombolysis in the treatment of early acute superior mesenteric artery embolization.Methods:The clinical data of 19 cases of acute superior mesenteric artery embolization treated with catheter directed thrombolysis therapy in the First Affiliated Hospital of Shandong First Medical University from Aug 2015 to Jul 2020 was retrospectively analyzed.Results:Fifteen cases were treated successfully with catheter directed thrombolysis , the blood flow and intestinal function restored significantly. The condition of 4 patients worsened during catheter directed thrombolysis therapy. Among them, three cases of intestinal necrosis were later confirmed by exploratory laparotomy and they recovered well after enterectomy. One elderly patient unsuitable for laparotomy died.Conclusion:Catheter directed thrombolysis is effective in the treatment of acute superior mesenteric artery embolization in early stage without intestinal necrosis.

Objective To investigate the outcome and affecting factors of patients with osteosarcoma after recurrence.Methods Retrospective analysis was used to analyze the achieved data and prognosis-related factors of 72 patients with extremity osteosarcoma after recurrence from June 2000 to September 2012 through multicenter joint treatment.37 males and 35 females were included.The overall age was 7 to 61 years old (average age22.2 years old).Classification based on the tumor recurrent parts:42 cases (58.3％) on femur,18 cases (25％) on tibia,7 cases (9.7％) on humerus and 5 cases (6.9％) on fibula.Classification based on metastases:14 cases of simple recurrence (19.4％),58 cases (80.6％) of recurrence with metastasis:29 cases (40.3％) of recurrence occurred first,9 cases (12.5％) of metastasis occurred firstly,20 cases (27.8％) of simultaneous recurrence with metastasis;12/14 patients (16.7％) of simple recurrence had long-term survival,but 2 died (2.8％).Six of 58 patients (8.3％) of recurrence with metastasis had long-term survival but 52 died (72.2％).Results The overall 10-year postrelapse survival rate of the 72 patients was 25％.Kaplan-Meier univariate analysis revealed that postrelapse survival was significantly influenced by the continued treatment after recurrence and the number of postoperative chemotherapy,but it was not significantlyinfluenced by age,gender and the number of preoperative chemotherapy.Multivariate analysis revealed that the continued treatment after recurrence and the number of postoperative chemotherapy were independent prognostic factors (P=0.002,P=0.007).At the same time,according to the survival curve grouped by different indicators,the number of chemotherapy after recurrence has no significant effect on TFS.However,the continued treatment after recurrence has obvious statistical significance on improving TFS and OS of patients (P=0.026,P=0.002).Conclusion Resection and standard postoperative chemotherapy had significant effects on the postrelapse survival in patients with osteosarcoma.There are good prospects for further multimodal therapy and multicenter cooperation for osteosarcoma after recurrence.