Impact of heterozygous ALK1 mutations on the transcriptomic response to BMP9 and BMP10 in endothelial cells from hereditary hemorrhagic telangiectasia and pulmonary arterial hypertension donors.

Heterozygous activin receptor-like kinase 1 (ALK1) mutations are associated with two vascular diseases: hereditary hemorrhagic telangiectasia (HHT) and more rarely pulmonary arterial hypertension (PAH). Here, we aimed to understand the impact of ALK1 mutations on BMP9 and BMP10 transcriptomic responses in endothelial cells. Endothelial colony-forming cells (ECFCs) and microvascular endothelial cells (HMVECs) carrying loss of function ALK1 mutations were isolated from newborn HHT and adult PAH donors, respectively. RNA-sequencing was performed on each type of cells compared to controls following an 18 h stimulation with BMP9 or BMP10. In control ECFCs, BMP9 and BMP10 stimulations induced similar transcriptomic responses with around 800 differentially expressed genes (DEGs). ALK1-mutated ECFCs unexpectedly revealed highly similar transcriptomic profiles to controls, both at the baseline and upon stimulation, and normal activation of Smad1/5 that could not be explained by a compensation in cell-surface ALK1 level. Conversely, PAH HMVECs revealed strong transcriptional dysregulations compared to controls with > 1200 DEGs at the baseline. Consequently, because our study involved two variables, ALK1 genotype and BMP stimulation, we performed two-factor differential expression analysis and identified 44 BMP9-dysregulated genes in mutated HMVECs, but none in ECFCs. Yet, the impaired regulation of at least one hit, namely lunatic fringe (LFNG), was validated by RT-qPCR in three different ALK1-mutated endothelial models. In conclusion, ALK1 heterozygosity only modified the BMP9/BMP10 regulation of few genes, including LFNG involved in NOTCH signaling. Future studies will uncover whether dysregulations in such hits are enough to promote HHT/PAH pathogenesis, making them potential therapeutic targets, or if second hits are necessary.

Differentiation of Crohn's disease, ulcerative colitis, and intestinal tuberculosis by dual-layer spectral detector CT enterography.

AIM: To investigate the value of radiological features and energy spectrum quantitative parameters in the differential diagnosis of Crohn's disease (CD), ulcerative colitis (UC), and intestinal tuberculosis (ITB) by dual-layer spectral detector computed tomography (CT) enterography (CTE). MATERIALS AND METHODS: Clinical and CTE data were collected from 182 patients with CD, 29 with UC, and 51 with ITB. CT images were obtained at the enteric phases and portal phases. The quantitative energy spectrum parameters were iodine density (ID), normalised ID (NID), virtual non-contrast (VNC) value, and effective atomic number (Z-eff). The area under curve (AUC) of the receiver operating characteristic curve (ROC) was calculated. RESULTS: The vascular comb sign (p=0.009) and enlarged lymph nodes (p=0.001) were more common in patients with CD than UC or ITB. In the differentiation of moderate-severe active CD from UC, enteric phase NID (AUC, 0.938; p<0.001) and portal phase Z-eff (AUC, 0.925; p<0.001) had the highest accuracy, which were compared separately. In the differentiation of moderate-severe active CD from ITB, enteric phase NID (AUC, 0.906; p<0.001) and portal phase Z-eff (AUC, 0.947; p<0.001) had the highest accuracy; however, the AUC value was highest when the four parameters are combined (AUC, 0.989; p<0.001; AUC, 0.986; p<0.001; AUC, 0.936; p<0.001; and AUC, 0.986; p<0.001). CONCLUSION: The present study shows that the combined strategies of four parameters have higher sensitivity and specificity in differentiating CD, UC, and ITB, and may play a key role in guiding treatment.

[Analysis of serum CA125 and related influencing factors in silicosis patients with pulmonary heart disease].

Objective: To analyze the serum carbohydrate antigen 125 (CA125) level and its influencing factors in male silicosis patients with pulmonary heart disease. Methods: In October 2021, data of 38 male patients with simple silicosis (silicosis group), 28 cases of silicosis with pulmonary heart disease (pulmonary heart disease group), and 27 healthy controls (control group) in the same age group were collected in inpatient and outpatient of Nanjing Occupational Disease Prevention and Control Hospital from January 2017 to December 2020. The serum CA125 levels of the three groups were compared, and the correlation between disease-related indexes and serum CA125 in silicosis patients with pulmonary heart disease was analyzed, as well as the influencing factors of pulmonary heart disease and serum CA125 levels in silicosis patients. Results: The serum CA125 level[ (19.95±7.52) IU/ml] in pulmonary heart disease group was higher than that in silicosis group[ (12.98±6.35) IU/ml] and control group[ (9.17±5.32) IU/ml] (P<0.05). There was no significant difference in serum CA125 level between the silicosis group and the control group (P>0.05). Serum CA125 levels were positively correlated with blood uric acid and fasting blood glucose in silicosis patients with pulmonary heart disease (r=0.39, 0.46, P<0.05). Serum CA125 level was a risk factor for silicosis patients with pulmonary heart disease (OR=1.13, 95%CI: 1.02-1.24, P<0.05). Dust exposure time, lactate dehydrogenase and smoking history were positively correlated with serum CA125 level in silicosis patients (P<0.05) . Conclusion: The serum CA125 level of male silicosis patients with pulmonary heart disease is significantly increased, and the level of CA125 is correlated with the level of fasting blood glucose and blood uric acid.

[Dysfunction of endothelial BMP-9 signaling in pulmonary vascular disease]. / Dysfonction de la signalisation endothéliale du BMP-9 dans les maladies vasculaires pulmonaires.

The signaling pathway of the bone morphogenetic protein (BMP)-9 binding to the endothelial receptor BMP receptor type II (BMPR-II), activin receptor-like kinase-1 (ALK1) and the coreceptor endoglin is essential to maintain the pulmonary vascular integrity. Dysregulation of this pathway is implicated in numerous vascular diseases, such as pulmonary arterial hypertension (PAH), hereditary hemorrhagic telangiectasia (HHT) and hepatopulmonary syndrome (HPS). This article aims to provide a comprehensive review of the implication of the BMP-9/BMPR-II/ALK1/endoglin pathway in the pathophysiology of these diseases.

CX3CR1 deficiency leads to impairment of immune surveillance in the epididymis.

Mononuclear phagocytes (MPs) play an active role in the immunological homeostasis of the urogenital tract. In the epididymis, a finely tuned balance between tolerance to antigenic sperm and immune activation is required to maintain epididymal function while protecting sperm against pathogens and stressors. We previously characterized a subset of resident MPs that express the CX3CR1 receptor, emphasizing their role in antigen sampling and processing during sperm maturation and storage in the murine epididymis. Bacteria-associated epididymitis is the most common cause of intrascrotal inflammation and frequently leads to reproductive complications. Here, we examined whether the lack of functional CX3CR1 in homozygous mice (CX3CR1EGFP/EGFP, KO) alters the ability of MPs to initiate immune responses during epididymitis induced by LPS intravasal-epididymal injection. Confocal microscopy revealed that CX3CR1-deficient MPs located in the initial segments of the epididymis displayed fewer luminal-reaching membrane projections and impaired antigen capture activity. Moreover, flow cytometry showed a reduction of epididymal KO MPs with a monocytic phenotype under physiological conditions. In contrast, flow cytometry revealed an increase in the abundance of MPs with a monocytic signature in the distal epididymal segments after an LPS challenge. This was accompanied by the accumulation of CD103+ cells in the interstitium, and the prevention or attenuation of epithelial damage in the KO epididymis during epididymitis. Additionally, CX3CR1 deletion induced downregulation of Gja1 (connexin 43) expression in KO MPs. Together, our study provides evidence that MPs are gatekeepers of the immunological blood-epididymis barrier and reveal the role of the CX3CR1 receptor in epididymal mucosal homeostasis by inducing MP luminal protrusions and by regulating the monocyte population in the epididymis at steady state as well as upon infection. We also uncover the interaction between MPs and CD103+ dendritic cells, presumably through connexin 43, that enhance immune responses during epididymitis. Our study may lead to new diagnostics and therapies for male infertility and epididymitis by identifying immune mechanisms in the epididymis.

Rotary table wobble error analysis and correction of a rotating accelerometer gravity gradiometer.

In a rotating accelerometer gravity gradiometer (RAGG), rotary table wobble refers to the shift in the direction of the spin axis during operation. This motion causes errors in the output of the RAGG, but the mechanism is not clear. The purpose of this paper is to analyze the relationship between rotary table wobble and RAGG errors and to propose a method for rejecting these errors. We consider the influence of attitude changes, angular velocity, and angular acceleration caused by the wobble on the specific force, and we describe the error transmission process based on the accelerometer configuration and its measurement principle. Furthermore, we show through a simulated experiment that when the angular velocity noise caused by the wobble is 1 µrad/s, this will produce errors of tens of E. We propose a post-error correction method that is based on the higher-precision RAGG model and motion measurement. The errors in the two channels of the RAGG are reduced to 3.69 E and 1.85 E after error correction. The error analysis of the effects of wobble on a RAGG and the proposed error correction method are of great significance for the development of high-precision gradiometers.

[Hepatopulmonary syndrome: Prevalence, pathophysiology and clinical implications]. / Syndrome hépatopulmonaire : prévalence, physiopathologie et implications cliniques.

Hepatopulmonary syndrome (HPS) is a pulmonary vascular disease induced by portal hypertension that is characterized by dilation of the pulmonary capillaries and right-to-left shunting, leading to gas exchange abnormalities. While dysregulation of several endothelial cell (EC)-derived angiocrine factors and the development of HPS have been associated, the field is currently lacking in mechanistic understanding on how they contribute to disease initiation, perpetuation, and worsening. Although substantial progress has been made in the description of clinical characteristics and outcomes, no specific targeting therapy has been shown to have a long-term effect on the evolution of HPS; only liver transplantation can lead full or partial reversibility of the syndrome. This article aims to provide a comprehensive review of the clinical and epidemiological definitions of HPS and to describe its experimental models as well as recent advances in understanding the pathophysiology of the disease, with specific focus on BMP-9 and its signaling pathway.

Comparing the efficacy of endovascular treatment for iliac vein compression syndrome with or without acute deep venous thrombosis: A single-center retrospective study.

BACKGROUND: To compare the efficacy of endovascular treatment for iliac vein compression syndrome (IVCS) with or without acute deep venous thrombosis of lower extremity. METHODS: This study retrospectively analyzed the clinical data of 300 IVCS patients, who received endovascular treatment between January 2013 and December 2017. According to whether IVCS was complicated by deep venous thrombosis or not, these patients were divided into non-thrombotic iliac vein lesion group (NIVL group, n = 127) and post-thrombotic iliac vein lesion group (PIVL group, n = 173). After endovascular treatment, all patients were followed up to assess the symptoms improvement and to evaluate the patency of iliac vein. RESULTS: The technical success rate was 98% (294/300), and percutaneous transluminal angioplasty with stenting was adopted in 294 cases. The incidence of perioperative complications was 36.33% (109/300), but no severe complications occurred. During a mean follow-up of 22.3 months (range 6-30 months), 9(6.82%, 9/132) patients in PIVL group had recurrence of deep venous thrombosis, but nobody had deep venous thrombosis and varicose veins recurrence in NIVL group. The effective rate of endovascular treatment in NIVL group and PIVL group was 96.88% and 90.15% (P = 0.050), while the cumulative primary patency of iliac vein in NIVL group was significantly higher than that in PIVL group (P = 0.008). CONCLUSIONS: The endovascular treatment is an effective, feasible, safe method for treating IVCS. There is no difference in the efficacy of IVCS patients with or without deep venous thrombosis, but the medium and long-term patency of patients with deep venous thrombosis is lower than that in patients without deep venous thrombosis.

[Artificial intelligence aided measurement of cervical squamous epithelial thickness and its correlation with cervical precancerous lesions].

Objective: To study the thickness of cervical squamous epithelia and its correlation with cervical precancerous lesions. Methods: We selected 495 HE slides of 209 cervical biopsies from January 2020 to June 2020 in the Department of Pathology, the First and Seventh Medical Center of the PLA General Hospital, including 173 slides with low grade squamous intraepithelial lesion (LSIL) and 214 slides with high grade squamous intraepithelial lesion (HSIL). Artificial intelligence labeling software was used to assist in measuring the epithelial thickness of normal cervical squamous epithelium, LSIL and HSIL of each slide. The thickest, thinnest, and middle widths of epithelial thickness were measured, respectively. Average epithelial thickness was defined as the sum of the above three widths divided by 3. The correlation statistical analysis was performed by combining the data of age and pathological diagnosis. Results: The average thickness of normal cervical squamous mucosa was (245.83±91.40) µm, which was (222.42±81.22) µm and was (195.95±66.59) µm in LSIL and HISL epithelial respectively (F=27.09, P<0.01). The average cell layers of normal cervical squamous epithelium was (15.5±4.2) layers, which of LSIL was (14.8±4.8) layers, and that of HSIL was (15.8±4.8) layers. The differences among normal, LSIL and HSIL were not statistically significant (P>0.05). Further statistical analysis was stratified by age (≤30 years, 31-40 years, 41-50 years, 51-60 years, and >60 years), the results of Pearson correlation analysis showed that the thickness of normal cervical squamous epithelial gradually thinned with age (correlation coefficient r=-0.141 9, P<0.05), while LSIL and HSIL epithelial thickness had significant correlation with age (P>0.05). In the subgroup of ≤50 years old, the epithelial thickness of normal squamous epithelium was the thickest, followed by LSIL, and HSIL epithelial thickness was the thinnest. The differences were statistically significant (P<0.05). While in the subgroup of >50 years, the differences were not statistically significant (P>0.05). Conclusions: The cervical squamous epithelium gradually becomes thinner with the degree of precancerous lesions increasing among patients of ≤50 years old. However, after age of 50 years, with the onset of menopause, the normal mucosal epithelium is becoming atrophy, so that mucosal thickness is no longer correlated with the extent of the lesion. In addition, it is suggested that the cervical vinegar white test performance during colposcopy is related to the protein changes in the mucosal epithelial cells, but not directly related to the thickness of the epithelial layer.

[Clinicopathological features and prognosis of gastrointestinal stromal tumor with PDGFRA-D842V mutation].

Objective: Platelet-derived growth factor alpha (PDGFRA) mutations are respectively rare in gastrointestinal stromal tumors (GIST). Most GIST with PDGFRA exon 18 mutations including D842V mutation are highly resistant to imatinib. The treatment of GIST harboring PDGFRA primary drug-resistant mutation is a major challenge. This article aims to investigate clinicopathologic features of GIST with PDGFRA-D842V mutation and the efficacy of comprehensive treatment, providing a reference for clinical practice. Methods: A retrospective cohort study was conducted to collect the clinicopathological and follow-up data of patients with GIST harboring PDGFRA mutation who were diagnosed and treated in the GIST Clinic of Renji Hospital from January 2005 to May 2020. According to the mutation site, the enrolled patients were divided into D842V mutation group and non-D842V mutation group. The differences of clinicopathologic characteristics between the two groups were compared. Furthermore, overall survival and prognostic factors were analyzed. Results: A total of 71 patients with PDGFRA-mutant GIST were included in this study, including 47 cases of D842V mutation (66.2%) and 24 cases of non-D842V mutation (33.8%). There were 28 male patients and 19 female patients in D842V mutation group, with a median age of 60 (36-82) years. There were 16 male patients and 8 female patients in non-D842V mutation group, with a median age of 62 (30-81) years. There were no significant differences in age, gender, primary location, surgical procedure, tumor size, mitotic count, expression of CD117 and DOG1, Ki-67 proliferation index and modified NIH grade between the two groups (all P>0.05). The positive rate of CD34 was 89.4% (42/47) and 62.5% (15/24) in the D842V mutation group and the non-D842V mutation group, respectively, with a statistically significant difference (χ(2)=5.644, P=0.018). Among all the cases, 66 cases underwent R0 resection without preoperative treatment; two cases underwent emergency operation with R1 resection because of tumor rupture; 2 cases were not operated after the pathological and mutation types were confirmed by biopsy (one case received avapritinib treatment and obtain partial remission). One case was diagnosed as wild-type GIST per needle biopsy in another institute, and underwent R0 resection after preoperative imatinib treatment for 6 months. After surgery, 5 high-risk GIST patients with D842V mutation and 5 high-risk GIST patients with non-D842V mutation were treated with imatinib for more than one year. The median follow-up time was 37 (1-153) months. As of the last follow-up among the patients who received R0 resection, 4 patients with D842V mutation had relapse, of whom 1 was in the period of imatinib administration, and the 3-year relapse-free survival rate was 94.2%; none of the patients with non-D842V mutation had relapse. There was no statistically significant difference in relapse-free surivval between two groups (P=0.233). Univariate analysis revealed that mitotic count (P=0.002), Ki-67 proliferation index (P<0.001) and modified NIH grade (P=0.025) were the factors associated with relapse-free survival of patients with D842V mutation after R0 resection (all P<0.05). However, the above factros were not testified as independant prognostic facors in multivariate Cox analysis (all P<0.05). Conclusion: Clinicopathologic features and the efficacy of radical resection in patients with PDGFRA-D842V mutation are similar to those in patients with non-D842V mutation.

[Research advances in the role of ubiquitin-proteasome pathway in hypertrophic scar].

The ubiquitin-proteasome pathway is a protein degradation pathway that relies on ATP and non-lysosomal pathway in eukaryotic cells. It participates in the regulation of multiple biological processes, including cell cycle, apoptosis, DNA repair, antigen presentation, receptor endocytosis, intracellular signal transduction. Recent studies have found that the ubiquitin-proteasome pathway can participate in the formation and development of hypertrophic scar by regulating transforming growth factor beta/Smad signal transduction and proliferation, differentiation, and apoptosis of fibroblasts. This article summarizes the effects of ubiquitin ligase enzyme, proteasome, and deubiquitinating enzyme in ubiquitin-proteasome pathway in hypertrophic scar, in order to provide new idea for the prevention and treatment of hypertrophic scar.

Diagnostic value of combined nucleic acid and antibody detection in suspected COVID-19 cases.

OBJECTIVES: Nucleic acid testing is the gold standard method for the diagnosis of coronavirus disease 2019 (COVID-19); however, large numbers of false-negative results have been reported. In this study, nucleic acid detection and antibody detection (IgG and IgM) were combined to improve the testing accuracy of patients with suspected COVID-19. STUDY DESIGN: The positive rate of nucleic acid detection and antibody detection (IgG and IgM) were compared in suspected COVID-19 patients. METHODS: A total of 71 patients with suspected COVID-19 were selected to participate in this study, which included a retrospective analysis of clinical features, imaging examination, laboratory biochemical examination and nucleic acid detection and specific antibody (IgM and IgG) detection. RESULTS: The majority of participants with suspected COVID-19 presented with fever (67.61%) and cough (54.93%), and the imaging results showed multiple small patches and ground-glass opacity in both lungs, with less common infiltration and consolidation opacity (23.94%). Routine blood tests were mostly normal (69.01%), although only a few patients had lymphopenia (4.23%) or leucopenia (12.68%). There was no statistical difference in the double-positive rate between nucleic acid detection (46.48%) and specific antibody (IgG and IgM) detection (42.25%) (P = 0.612), both of which were also poorly consistent with each other (kappa = 0.231). The positive rate of combined nucleic acid detection and antibody detection (63.38%) was significantly increased, compared with that of nucleic acid detection (46.48%) and that of specific antibody (IgG and IgM) detection (42.25%), and the differences were statistically significant (P = 0.043 and P = 0.012, respectively). CONCLUSIONS: Nucleic acid detection and specific antibody (IgG and IgM) detection had similar positive rates, and their combination could improve the positive rate of COVID-19 detection, which is of great significance for diagnosis and epidemic control.

[Impacts of high-voltage electrical burn on serum platelet-related factors and platelet aggregation number in rats and the interventive effect of Xuebijing].

Objective: To explore the effect of high-voltage electrical burn on platelet function and rheological behavior in rats and the interventive effect of Xuebijing. Methods: A total of 280 Sprague Dawley rats of clean grade (aged 8-10 weeks, male and female unlimited) were divided into sham injury group, simple electrical burn group, electrical burn+ saline group, and electrical burn+ Xuebijing group according to the random number table, with 70 rats in each group. Rats in sham injury group were not conducted with electrical current to cause sham injury. Rats in the other three groups were given electrical current with output voltage of 2 kV and current intensity of (1.92 ± 0.24) A for 3 s, which caused high-voltage electrical burn wounds, each with an area of 1 cm×1 cm distributed in the left forelimb at the current inlet and the right hindlimb at the current outlet respectively. Rats in sham injury group and simple electrical burn group were not treated after injury. At post injury minute 2 and on post injury day (PID) 1, 2, 3, 4, 5, and 6, rats in electrical burn+ saline group and electrical burn+ Xuebijing group were intraperitoneally injected with 6 mL/kg saline and 6 mL/kg Xuebijing, respectively. Survival conditions of rats were recorded during the experiment. At 15 min before injury and at post injury hour (PIH) 1, 8, 24, 48, 72, and on PID 7, 10 rats in each group were respectively selected according to the random number table to sacrifice after collection of 5 mL blood under the direct vision of heart. Blood in the volume of 0.05 mL from each rat was taken to make blood smear, and platelet aggregation number was counted under 400 fold field of view using multiple projection microscope. The remaining blood samples were centrifuged to collect supernatant, and the content of platelet-derived growth factor (PDGF), thrombopoietin (TPO), and platelet activating factor (PAF) was detected by enzyme-linked immunosorbent assay. Data were statistically analyzed with analysis of variance for factorial design and Student-Newman-Keuls method. Results: All rats in sham injury group and simple electrical burn group survived during the experiment. One rat in electrical burn+ saline group died on PID 6, and one rat on PID 5 and one rat on PID 6 died in electrical burn+ Xuebijing group. The levels of all indexes among the 4 groups were close at 15 min before injury. The serum content of PDGF, TPO, and PAF and platelet aggregation number of rats in the three electrical burn groups at all time points after injury were higher or more than those in sham injury group, and the first three indexes reached the peak at PIH 8. The serum platelet aggregation number of rats in simple electrical burn group reached the peak at PIH 48, and that in electrical burn+ saline group and electrical burn+ Xuebijing group reached the peak at PIH 72. Among them, the serum content of PDGF of rats in electrical burn+ Xuebijing group at PIH 48, 72 and on PID 7 ((12.8±4.0), (11.6±4.4), (11.0±3.6) ng/mL, respectively) was close to that in sham injury group ((10.4±2.0), (10.4±2.5), (9.8±3.3) ng/mL, respectively, P>0.05). The serum content of TPO of rats in electrical burn+ Xuebijing group at PIH 24, 72 and on PID 7 ((200±52), (192±36), (193±32) ng/mL, respectively) was close to that in sham injury group ((182±30) , (184±41), (183±33) ng/mL, respectively, P>0.05). The serum content of PDGF, TPO, and PAF and platelet aggregation number of rats in electrical burn+ Xuebijing group at every time point after injury was generally lower or less than that in electrical burn+ saline group and simple electrical burn group. Conclusions: Application of Xuebijing treatment after high-voltage electrical burn can decrease the content of PDGF, TPO, and PAF in the serum and reduce the number of platelet aggregation, thereby inhibit platelet activation and improve platelet rheology.

[Effects and mechanism of rat epidermal stem cells treated with exogenous vascular endothelial growth factor on healing of deep partial-thickness burn wounds in rats].

Objective: To explore the effects and mechanism of rat epidermal stem cells (ESCs) that were treated with exogenous vascular endothelial growth factor (VEGF) on the healing of deep partial-thickness burn wounds in rats. Methods: ESCs were isolated and cultured from the trunk skin of a 3-month-old female Sprague-Dawley (SD) rat. The third passage of cultured cells in the logarithmic growth phase was used in experiments (1)-(3). (1) The cells were routinely cultured in keratinocytes-specified serum-free medium (K-SFM) (the same routine culture condition below). The morphology of cells cultured for 3 and 5 days was observed under the inverted optical microscope. (2) After 24 hours in routine culture, the expression of cell surface markers CD44, CD45, CD11b, and CD11c was detected by flow cytometer, with triplicate samples. (3) Four batches of cells were collected, and each batch was divided into VEGF group or blank control group according to the random number table. The cells in blank control group were routinely cultured, while the cells in VEGF group were cultured in K-SFM containing VEGF in the final mass concentration of 10 ng/mL. The protein expressions of cytokeratin 19 (CK19) and CK10 in cells cultured for 10 days were detected by Western blotting. The Nanog mRNA expression in cells cultured for 0 (immediately), 2, 4, 6, 8, and 10 day (s) was detected by real-time fluorescent quantitative reverse transcription polymerase chain reaction. The absorbance value was detected with cell counting kit-8 in cells cultured for 2, 4, 6, 8, and 10 days. The clone number of more than 50 cells was observed and counted under the optical microscope in cells cultured for 10 days, and the cell colony formation rate was calculated. Three samples at each time point was analysed. (4) Thirty-six 3-month-old SD rats (either male or female) were used for the study, and two deep partial-thickness burn wounds (10 mm in diameter) were created in each rat by pressing a 100 ℃ electric iron plate on symmetric dorsal side. According to the random number table, the injured rats were divided into VEGF+ ESCs group, ESCs alone group, and blank control group, with 12 rats and 24 wounds in each group. From 0 (immediately) to 2 day (s) after injury, 20 µL phosphate buffer solution (PBS) was injected into each wound in the three groups in one time, once a day, with the solution in VEGF+ ESCs group containing 0.8×10(6) cells/mL of ESCs treated by 10 ng/mL VEGF for 10 days, the solution in ESCs alone group containing 0.8×10(6) cells/mL of ESCs without any treatment, and the solution in blank control group being PBS only. On post first injection day (PFID) 0 (immediately), 3, 7, and 14, three rats from each group were taken respectively according to the random number table for wound healing assessment, and the wound healing rates on PFID 3, 7, and 14 were calculated. The mice at each time point were sacrificed with wound tissue harvested for histology, and the skin structure was observed by hematoxylin-eosin staining. Data were statistically analyzed with independent sample t test, analysis of variance for factorial design, least significant difference test, and Bonferroni correction. Results: (1) By day 3 in culture, cells distributed in slowly-growing clusters. By day 5, the clusters were large and round, in which the cells mainly with large and round nuclei and little cytoplasm were observed. The above results were consistent with the morphological characteristics of ESCs. (2) The positive expression rate of CD44 was (94.3±1.2) %, and the expressions of CD45, CD11b, and CD11c were negative. The cells were confirmed as ESCs. (3) Compared with those of blank control group, the protein expression of CK19 in the cells of VEGF group was significantly increased after 10 days in culture (t=3.756, P<0.05), while the protein expression of CK10 was significantly decreased (t=3.149, P<0.05). Compared with those of blank control group, the Nanog mRNA expression in the cells cultured for 0 and 2 day (s) and absorbance values of the cells cultured for 2 and 4 day (s) were not significantly changed in VEGF group (t=0.58, 0.77, 0.53, 3.02, P>0.05), while the Nanog mRNA expression in the cells cultured for 4, 6, 8, and 10 days and absorbance values of the cells cultured for 6, 8, and 10 days were significantly increased in VEGF group (t=6.34, 5.00, 5.58, 4.61, 5.65, 10.78, 15.51, P<0.01). After 10 days in culture, the cell colony-forming rate in VEGF group was (56.4±1.3) %, significantly higher than (31.5±1.3) % of blank control group (t=13.96, P<0.01). (4) The burn wounds of rats in the three groups were confined to the superficial dermis of the skin on PFID 0. On PFID 3, normal skin tissue at wound margins slightly contracted in the rats of VEGF+ ESCs group, which was earlier than that in the other two groups. On PFID 7, the newly generated epidermis covered most parts of the rat wounds in VEGF+ ESCs group, and some of the epithelium crawled around the rat wounds in ESCs alone group, but no obvious epithelialization was observed in the rat wounds in blank control group. On PFID 14, the rat wounds in VEGF+ ESCs group were basically healed, while some parts of the rat wounds were unhealed in ESCs alone group, and most parts of the rat wounds were unhealed in blank control group. On PFID 3, the wound healing rates of rats in the three groups were similar (P>0.05). On PFID 7 and 14, the wound healing rates of rats in ESCs alone group, respectively (26.0±2.0) % and (64.4±4.7) %, were obviously higher than (12.4±1.1) % and (29.1±3.3) % of blank control group (P<0.01), all of which were obviously lower than (41.0±2.4) % and (91.3±3.5) % of VEGF+ ESCs group (P<0.01). On PFID 3, infiltration of a large number of inflammatory cells were observed in the rat wounds in VEGF+ ESCs group, which was earlier than those in the other two groups. On PFID 7, a large number of endothelial cells were observed in the rat wounds in VEGF+ ESCs group, while proliferation of a few endothelial cells were observed in the rat wounds in ESCs alone group, and a large number of inflammatory cells infiltrated the rat wounds in blank control group. On PFID 14, the newly generated epidermal cells covered nearly all the rat wounds in VEGF+ ESCs group and most parts of the rat wounds in ESCs alone group, while a large number of endothelial cells were observed and the newly generated epidermal cells covered some parts of the rat wounds in blank control group. Conclusions: ESCs of rats treated with exogenous VEGF can promote the healing of deep partial-thickness burn wounds in rats, which may be related to VEGF's roles in promoting the proliferation of ESCs and reducing its differentiation level, so as to maintain the potency of stem cells.

[Application of imatinib plasma concentration monitoring in the whole process management of gastrointestinal stromal tumor patients].

Objective: To investigate the significance of monitoring imatinib mesylate (IM) plasma concentrations in patients with gastrointestinal stromal tumor (GIST). Methods: A retrospective descriptive study was carried out. Inclusion criteria: (1) patients with GIST confirmed by postoperative pathology or puncture pathology receiving maintenance therapy of IM; (2) administration of same dose of IM for at least 4 weeks (achieving steady - state plasma concentration). Patients who had severe organ dysfunction, received IM generics, or received IM simultaneously with other drugs significantly affecting IM pharmacokinetic were excluded. A total of 185 patients at the GIST Clinic of Renji Hospital, Shanghai Jiaotong University School of Medicine from August 2018 to May 2019 were enrolled, including 114 males (61.6%) and 71 females (38.4%) with a median age of 60 years old (range, 30-89 years), and 63 advanced cases. Patients receiving preoperative or postoperative adjuvant therapy were given IM 400 mg QD; patients with KIT exon 9 mutation or with disease progression during IM 400 mg QD treatment were given IM 600 mg QD. If the patient had adverse reactions such as myelosuppression during the medication, IM would be reduced or given BID per day. The peripheral venous blood was collected (22 to 24 hours after the last dose for patients who took IM QD and 2 hours before the first dose per day for those who took IM BID). IM plasma concentration was measured through high performance liquid chromatography coupled with tandem mass spectrometry (HPLC-MS/MS). Correlation analysis between IM plasma concentration results and clinical data was performed using linear regression analysis. Results: A total of 241 stable blood samples of IM plasma concentration from 185 patients were finally collected. The IM plasma concentrations were significantly different between the doses of 300 mg/d and 400 mg/d [(942.4±433.5) µg/L vs. (1340.0±500.1) µg/L, t=6.317, P<0.001], and between 400 mg/d and 600 mg/d [(1340.0±500.1) µg/L vs. (2188.0±875.5) µg/L, t=3.557, P=0.004]. Among the blood samples of 57 patients receiving IM 300 mg/d, the IM plasma concentration of the advanced patients was significantly lower than that of the non-advanced patients [(795.6±225.8) µg/L vs. (992.2±484.4) µg/L, t=2.088, P=0.042]. Among the 137 blood samples of patients receiving IM 400 mg/d, the IM plasma concentration was higher in patients aged >60 years than those aged ≤60 years [(1461.0±595.3) µg/L vs. (1240.0±380.9) µg/L, t=2.528, P=0.013] and the IM plasma concentration of cases with diarrhea was significantly lower than that of those without diarrhea [(745.8±249.6) µg/L vs. (1382.0±486.9) µg/L, t=6.794, P<0.001]. Gender, primary location, surgical procedure, mutated gene, mutation type, or time of administration was associated with IM plasma concentration no matter in patients taking IM doses of 400 mg/d or 300 mg/d (all P>0.05). Regression analysis showed that body mass (P=0.004 and P=0.019), body mass index (P=0.016 and P=0.042), and body surface area (P=0.007 and P=0.028) were all negatively correlated with IM plasma concentrations in patients taking IM doses of 300 mg/d and 400 mg/d. Within the 137 patients who received a fixed oral dose of 400 mg/d IM, 17 patients received oral 200 mg BID, whose IM plasma drug concentration was not significantly different compared with that of 120 patients who received 400 mg IM QD [(1488.0±408.3) µg/L vs. (1319.0±509.7) µg/L, t=1.307, P=0.193]. Conclusions: Monitoring IM plasma concentration is significant throughout the whole process of management of GIST patients receiving IM treatment. In particular, regular monitoring IM plasma concentration and developing appropriate treatment strategies can bring better therapeutic benefits for patients with low doses, diarrhea, advanced condition and older age.

[Epidemiological characteristics and antibiotic resistance of Campylobacter spp. among diarrhea outpatients in Shanghai, 2013-2016].

Objective: To understand the epidemiological characteristics and antibiotic resistance profiles of Campylobacter spp. in Shanghai from 2013 to 2016. Methods: Stool samples collected from diarrhea outpatients were cultured for Campylobacter spp., using the membrane filter method in 23 hospitals under the sentinel programs, from 2013 to 2016. All the strains were identified by biochemical tests and PCR. Broth microdilution method was used to investigate the antibiotic resistance of 179 Campylobacter spp. strains that including azithromycin, ciprofloxacin, erythromycin, gentamicin, tetracycline, nalidixic acid, telycin, klinthromycin and flurbenicol. Results: A total of 179 Campylobacter spp. strains were isolated from 10 444 stool samples (1.7%). Campylobacter jejuni and Campylobacter coli appeared as the predominant ones (94.4% and 5.6%). The incidence rate was higher in children than that in adults, with peaks of infections mainly from April to June and October to December. Campylobacter jejuni strains seemed highly resistant to ciprofloxacin (96.4%), tetracycline (83.4%) and nalidixic acid (81.7%). The resistant rates appeared higher on Campylobacter coli strains that isolated from patients. Some strains were resistant to multi-drugs. Conclusions: Campylobacter spp. seemed one of the important pathogens that isolated from outpatients with diarrhea, in Shanghai. Both age and season related characteristics of Campylobacter spp. were seen. Campylobacter spp. isolated from patients was highly resistant to ciprofloxacin, tetracycline and nalidixic acid.

[Investigation and analysis of thyroid function of medical occupational population in a tertiary-A hospital in Nanjing].

Objective: To evaluate the effect of low-dose of ionizing radiation on thyroid function of medical occupational group with long-term exposure, furthermore, to analyze the relationship between the thyroid hormones and the risk factors, such as exposure length, department and comulative radiation dose. Ultimately, providing the scientific basis for setting the ionizing radiation protection standards. Methods: The population who engaged radiodiagnosis and radiotherapy in a tertiary-A hospital was set up as occupational exposure, and the administrative staffs in a company were considered as control. According to the inclusion and exclusion criteria, 161 medical professionals and 159 administrative staffs as the research object.We figured out the basic information and general condition of the groups by face-to-face questionnaire survey, calculated the annual comulative radiation dose through local center for disease control and prevention, By means of the thyroid hormone testing, we analyzed the thyroid hormone levels with different population, occupational exposure factors. Applying Epidata、Excel in data management. All the data was analyzed by statistical software package Stata12.0. Descriptive statistics, single factor analysis of variance and other statistical methods were used for data analysis. Test standard: α=0.05、P<0.05 statistical significant. Results: Age, sex and seniority were proportionality between exposure and control groups. The dosages of occupational population exposure to ionizing radiation were about 1/10 of national permit value, belonging to low-dose exposure. The T(3), FT(3) level of the exposure group was decreased than the control group (P<0.001). especially the FT(3) level has statitical discrepancy among groups with different exposure length (P<0.05). Conclusion: Long-term exposure to low-dose ionizing radiatiom can induce the thyroid damage of medical occupational population, which should be broader concerned.

[New insights in the pathogenesis of pulmonary arterial hypertension]. / Nouveau regard sur la physiopathologie de l'hypertension artérielle pulmonaire.

Pulmonary arterial hypertension (PAH) is a severe and incurable cardiopulmonary disorder. Research from the past 10 years illustrates the complex and multifactorial aspects of PAH pathophysiology. Furthermore, latest advances in the field have led to a better understanding of the key components underlying this inadequate accumulation of pulmonary vascular cells within the pulmonary arterial walls, leading to pulmonary vascular remodelling. Among the underlying molecular and cellular mechanisms, pulmonary endothelial dysfunction, alterations of the inter-cell communications within the pulmonary arterial walls as well as defects of the inflammatory component and the loss of BMPRII activity play critical roles in the pathogenesis of the disease.