OBJECTIVE To analyze the influential factors on trough concentration （cmin） and area under the drug concentration time curve （AUC） of voriconazole （VRZ） in pediatric patients with thalassemia undergoing hematopoietic stem cell transplantation （HSCT）. METHODS A total of 60 pediatric patients with thalassemia undergoing HSCT who used VRZ for prevention or treatment of invasive fungal infection were collected in our hospital from January 2021 to January 2024. The plasma concentration of VRZ was measured by high-performance liquid chromatography and the AUC was calculated. The factors affecting cmin and AUC of VRZ were analyzed using multiple linear regression. RESULTS A total of 120 cases of VRZ cmin in 60 pediatric patients was obtained and 27 cases of VRZ AUC in 26 pediatric patients were obtained. The median concentration of VRZ cmin was 0.31 mg/L； 46 cases had a cmin in 0.5-5 mg/L（ 38.33%）， 2 cases had a cmin＞5 mg/L（ 1.67%）， and 72 cases had a cmin＜0.5 mg/L. The median AUC of VRZ was 11.68 mg·h/L. The patient’s body weight， HSCT postoperative days， lymphocyte count， and combined use of phenytoin sodium， tacrolimus or cyclosporine had significant effects on VRZ cmin （P＜0.05）. Lymphocyte count and combined use of phenytoin sodium had significant effects on VRZ AUC （P＜0.05）. CONCLUSIONS The body weight， HSCT postoperative days， lymphocyte count， and combined use of phenytoin sodium， tacrolimus or cyclosporine are independent factors affecting VRZ cmin. Lymphocyte count and combined use of phenytoin sodium are independent factors affecting VRZ AUC.

Objective:To investigate the role of protein phosphatase 4 catalytic subunit (PP4C) in regulating hepatitis B virus X protein (HBx) levels and its effects on the biological functions of HBx, thus to provide a potential therapeutic targets for hepatitis B virus (HBV)-related hepatocellular carcinoma.Methods:In vivo and in vitro interactions between HBx and PP4C were analyzed by co-immunoprecipitation (Co-IP) and GST pull-down assay. Recombinant plasmids of PP4C and HBx were co-transfected with Lipofectamine 3000 reagents into hepatoma cells to detect the protein levels of HBx by Western blot. The half-life of HBx in the transfected cells treated with cycloheximide (CHX) were detected. The phosphorylation assay was used to evaluate the effects of PP4C on HBx phosphorylation. CCK8 assay, wound healing assay and Matrigel invasion chamber assay were used to analyze the effects of PP4C on the biological functions of HBx. Results:PP4C interacted with HBx in vivo and in vitro. PP4C overexpression significantly increased the protein level and stability of HBx and the phosphorylation assay confirmed that PP4C overexpression decreased the serine phosphorylation of HBx in hepatoma cells. PP4C overexpression enhanced the migration and invasion of hepatoma cells, but had no significant effects on the proliferation. Conclusions:The interactions between HBx and PP4C promoted the stability of HBx and ultimately enhanced the migration and invasion of hepatoma cells, and the mechanisms might be related to the decrease of HBx serine phosphorylation by PP4C. This study provided a theoretical basis for further investigation of the pathogenic mechanisms of HBx, and targeting PP4C and HBx interaction might provide insights for developing novel treatment for HBV-related hepatocellular carcinoma.

Objective:To investigate the expression level of miR-21 in patients with septic acute respiratory distress syndrome, and analyze its correlation with inflammatory factors and its value in evaluating clinical prognosis.Methods:Patients with sepsis admitted to the intensive care unit (ICU) of the First Affiliated Hospital of Hebei North University from October 2020 to March 2022 were prospectively recruited. The patients were divided into an ARDS group and a non-ARDS group according to whether ARDS occurred within 3 days after admission. The ARDS group was divided into a low-risk group and a high-risk group according to the oxygenation index, and the ARDS group was divided into a survival group and a death group according to their 28-day survival status. In addition, 50 healthy volunteers who underwent physical examination during the same period were included in the control group. The expression levels of serum miR-21 and inflammatory factors [tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6)] were compared in each group at admission. To analyze the relationship between miR-21 expression and inflammatory factors and explore the value of miR-21 in predicting the prognosis of sepsis patients with ARDS.Results:A total of 106 patients with sepsis were included. Compared with the healthy control group, the levels of serum miR-21, TNF-α and IL-6 in the ARDS group and non-ARDS group were significantly increased (all P<0.05). Compared with the low-risk group, the levels of serum miR-21, TNF-α and IL-6 in the high-risk group were significantly increased (all P<0.05). Compared with the survival group, the serum miR-21 expression level and TNF-α and IL-6 levels in the death group were significantly increased (all P<0.05). Pearson correlation analysis showed that the serum miR-21 expression level in the ARDS group was positively correlated with IL-6 and TNF-α levels ( r=0.842, 0.697, P<0.001). ROC curve analysis showed that the AUC of serum miR-21 in predicting death from sepsis complicated with ARDS was 0.876 (95% CI: 0.761-0.942), the best cut-off value was 2.25, and its sensitivity and specificity were 92.9% and 71.7%, respectively. Conclusions:The expression level of serum miR-21 is significantly increased in patients with sepsis ARDS, is positively correlated with the level of inflammatory factors, is closely correlated with the severity and prognosis of the disease, and has great value in evaluating the prognosis of patients with sepsis ARDS.

Objective:To observe the effect of exogenous hydrogen sulfide(H 2S) on the proliferation of rabbit vascular smooth muscle cell(VSMC) under high static pressure. Methods:Rabbit thoracic aorta VSMC were isolated and cultured under high static pressure(100mmHg) and divided into control group [cultured with 0.2%fetal bovine serum(FBS) and no NaHS]、10%FBS group(10%FBS and no NaHS) and NaHS group(10%FBS and 50mmol/L NaHS). VSMC proliferation was analyzed with BUDR. CSE, Calmodulin(p-CaM)and CyclinD1 protein levels of VSMC were measured by Western blotting.Results:Compared with the 10%FBS group, NaHS inhibit the proliferation of rabbit VSMC significantly (0.50±0.03 vs. 0.26±0.03, P<0.05). Compared with control group, CSE protein in the 10%FBS group decreased significantly(1.21±0.10 vs. 0.33±0.04, P<0.05) and p-CaM and CyclinD1 protein increased significantly(0.23±0.04 vs. 0.86±0.04 and 0.22±0.03 vs. 1.19±0.06, P<0.05). Compared with the 10%FBS group, CSE protein in NaHS group increased significantly(0.33±0.04 vs. 1.11±0.11, P<0.05), and the expression of p-CaM and CyclinD1 protein decreased significantly(0.86±0.04 vs. 0.26±0.05 and 1.19±0.06 vs. 0.51±0.03, P<0.05). Conclusion:Exogenous hydrogen sulfide inhibits the proliferation of VSMC under high static pressure by the CSE/H2S pathway which related to the reduction of the expression of p-CaM and CyclinD1.

Objective To report the clinical management of the first case of human infection with influenza A (H7N4) in the world ,and to explore the clinical characteristics and therapeutic regimen of H7N4 infection in humans .Methods A case of confirmed human infection with influenza A (H7N4) virus was hospitalized in the Third People′s Hospital of Changzhou in Jiangsu Province on January 1st ,2018 . The clinical characteristics and treatment were analyzed by reviewing the clinical data and management of the patient ,and compared with the clinical characteristics of human infection with H 7N9 cases .Results The patient was an elderly woman with hypertension ,who had an exposure history of live poultry before onset .The patient had flu-like symptoms including fever , cough and expectoration . The symptoms worsened in the fifth day and presented as respiratory failure and circulatory disturbance .The count of white blood cells ,lymphocytes and platelets counts declined ,while the levels of aspartate aminotransferase and c-reactive protein slightly increased .The levels of creatine kinase and lactate dehydrogenase deformity increased .The pulmonary lesions manifested as exudation and consolidation of both side in a short period of time after the onset . Treatment was initiated according to the epidemiology history , clinical manifestations ,laboratory and imaging characteristics . The primary diagnosis was human infection of avian influenza virus .The patient was treated with oxygen therapy ,antivirus ,anti-infection and short-duration corticosteroids treatment . Early and timely throat swab specimens were tested . The patient recovered and discharged from hospital after active treatment .It was confirmed by the Chinese Center for Disease Control and prevention that the patient was infected with avian influenza A (H7N4) virus ,and no close contact was infected .Conclusions There are many similarities of clinical features between case of human infection with H7N4 and that of H7N9 . The early treatment of neuraminidase inhibitors is recommended according to the diagnosis and treatment of human infection with H 7N9 .In the clinical practice ,the avian influenza nucleic acid should be tested in time to determine the cause of the disease in those patients with suspected viral pneumonia ,especially in those who has an avian contact history .

In the development of cancer, the DNA fragments of tumor cells enter into the blood circulation to form circulating tumor DNA (ctDNA), which is an indicator for detecting and quantifying tumor mutations. Epithelial ovarian cancer accounts for most of the histological types in ovarian cancer. This article reviews the clinical application of ctDNA in epithelial ovarian cancer, including the early diagnosis, therapeutic effect evaluation after operation and adjuvant therapy, tumor load assessment, dynamic monitoring during the follow-up and the prognosis of patients.

Objective To analyze the serum samples of young and middle-aged patients with hypertension of different syndromes and healthy patients by means of metabonomics; To confirm its biomarker; To reveal the nature of phlegm dampness retention syndrome in essential hypertension. Methods The syndrome types of 34 hypertension patients were differentiated into 14 cases of liver-fire hyperactivity syndrome group, 14 cases of phlegm dampness retention syndrome group and 6 cases of yin deficiency and yang hyperactivity syndrome group. 15 healthy volunteers were selected as control group. 1H-NMR technique combined with partial least squares-discriminate analysis (PLS-DA) method was used to look for the biomarkers. And the levels of UA, TC and TG were recorded. Results The level of UA in phlegm dampness retention syndrome group was significantly higher than the other three groups (P<0.05). The level of TC in yin deficiency and yang hyperactivity syndrome group was higher and had significantly difference with the control group (P<0.05). The level of TG in phlegm dampness retention syndrome group was higher and had significantly difference with control group (P<0.05). The metabonomics study based on the 1H-NMR method could distinguish the phlegm dampness retention syndrome group from the normal control group, liver-fire hyperactivity syndrome group and yin deficiency and yang hyperactivity syndrome group. Compared with the control group, the levels of acetone, VLDL, and LDL were significantly higher and the levels of lactate, serine, glucose, methionine, alanine were significantly lower in phlegm dampness retention syndrome group; compared with the liver-fire hyperactivity syndrome group, levels of citrate, alanine, VLDL, and LDL were significantly higher and the levels of glucose, lysine, glutamate, proline lactate were significantly lower; compared with yin deficiency and yang hyperactivity syndrome group, the level of creatinine was significantly higher. Conclusion There are lipoprotein metabolism, amino acid metabolism and glucose metabolism disorder in the young and middle-age hypertensive patients with phlegm dampness retention syndrome.

Clostridium cellulolyticum, as one of obligate anaerobic bacteria capable of secreting cellulosome, has not been efficiently cultured due to its strict requirement of growing conditions. In this study, culture conditions of C. cellulolyticum were optimized using response surface methodology. Plackett-Burman design was first used to screen the dominant impact factors for the growth of C. cellulolyticum, which were determined as yeast extract concentration, cellobiose concentration and culture temperature. The steepest ascent path design was then applied to gain the suitable range close to the optimal culture conditions for obtaining high cell density. The central composite design and the response surface analysis were finally used to determine the optimal levels of the influential factors, which were 3 g/L for yeast extract concentration, 7 g/L cellobiose concentration and 34 degrees C for culture temperature. The optimized medium was used for flask culture, and OD600 of C. cellulolyticum was increased from 0.303 to 0.586. With a pH-controlled fermentor at batch mode, OD600 reached 3.432, which was 2.8 times higher than elsewhere reported. These results support further study on the high-density culture of C. cellulolyticum and its application.
Bacteriological Techniques ; methods ; Clostridium cellulolyticum ; growth & development ; Culture Media ; Population Density

Objective To observe the effect of Ruanmailing Oral Liquid on angiogenesis in atherosclerosis plaque of Apolipoprotein E (ApoE) gene knock-out mice, and explore the mechanisms of plaque stabilizing. Methods Totally 30 mice 6-8 weeks old ApoE knockout mice were fed a high fat diet for 12 weeks until the formation of a mature atherosclerotic plaque. They were randomly divided into three groups-model group, Ruanmailing group, simvastatin group, with another 6 normal C57BL/6J mice as the control group, and were administered for 12 weeks. Blood was extracted from orbital venous to measure the lipid (TC, TG, LDL-C, HDL-C) variation. HE-stain was used to observe aortic pathomorphological changes, meanwhile, immunohistochemical method was adopted to determine the microvessel density of plagues which is marked by CD105, as well as the expression of CD105 in aorta. Results Compared with the model group, TC, LDL-C, TG of Ruanmailing group and simvastatin group decreased significantly, while HDL-C increased significantly (P<0.01), degree of pathological damage was reduced, microvessel density and the expression of CD105 was significantly lower (P<0.01). There was no significant difference between Ruanmailing group and simvastatin group in above-mentioned indicators (P>0.05). Conclusion Ruanmailing Oral Liquid may reduce the expression of CD105 and inhibit the angiogenesis within the plaque, which is the possible mechanism of stabilizing the atherosclerotic plaque.

Objective To investigate the effect of Kangxin Capsule on the stability of atherosclerotic plaque in rabbits. Methods Forty purebred male New Zealand white rabbits were randomly divided into model group, atorvastatin group, Kangxin Capsule high and low dose groups, and normal control group, 8 rabbit for each group, and were given corresponding administration. Atherosclerosis model was made by intimal injury and high cholesterol diet except for normal control group. After 6 W, aortic vascular pathomorphology were observed, and immunohistochemical method was used to detect the expression of MMP-2 and VCAM-1 in atherosclerotic plaques. Results Compared with model group, the ratio of the thickness of wall to the area of lumina (W/L), average optical density, and the percentage of positive area of Kangxin Capsule groups were less, also, the expression of MMP-2 and VCAM-1 were reduced. The differences were significant (P<0.05 or P<0.01), especially in the high-dose group. Conclusion Kangxin Capsule can reduce the expression of MMP-2 and VCAM-1, which is the possible mechanism of stabilizing the atherosclerotic plaque.