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.

[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.

[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.

[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.

[Effect of aspirin on breast cancer stem cells and stemness of breast cancer].

Objective: To explore the effect of aspirin on the stemness of breast cancer cells and apoptosis induction of breast cancer stem cells. Methods: The 4T1 cells cultured with stem cell culture medium were screened, and immunofluorescence technique, flow cytometry and tumor-forming experiment in vivo were applied to test stem cell characteristics of the tumor spheres. After dealt with aspirin, the apoptosis rate of 4T1 stem cells was analyzed by flow cytometry. The 4T1 cells were cultured in vitro and treated with aspirin, then flow cytometry analysis was used to detect the expression of aldehydedehy drogenase1 (ALDH1), and the expression of stemness genes was tested by Western blot . Then, after culturing the cells with medium containing basic fibroblast growth factor (bFGF), epidermal growth factor (EGF), B27 and N2, the ability of sphere-forming was observed and recorded by microscopy. In vivo BALB/c mice inoculated with 4T1 stem cells were randomly divided into the control group, 10 mg/kg, 30 mg/kg and 100 mg/kg aspirin groups. After 10 days, the mice were dealt with aspirin or NS for 15 days, then the tumor growth was observed and recorded. Results: The ratio of ALDH1 positive cells was up to 78.55%, and 4T1 tumor sphere had a postive expression of ALDH1 and sex determining region Y-box 2 (SOX2). In vivo tumorigenesis abilities of tumor sphere with 1×10(2) 4T1 stem cells could be 75%, while the ratio of normal cells was zero. The ratio of Aspirin-induced apoptosis of 4T1 stem cells at early stage and and late stage increased from 0.36% to 21.61%, and from 4.21% to 21.38%, respectively. Flow cytometry and Western blot assay results indicated that aspirin could reduce the expression of ALDH1, SOX2, octamer-binding transcription factor 4 (OCT4) and NANOG in 4T1 cells. Sphere-forming experiments results showed that aspirin could inhibit sphere forming ability of breast cancer cells. In vivo, aspirin inhibited the growth of tumors with a dose-dependent manner. Conclusion: Aspirin could induce apoptosis of cancer stem cells and reduce stemness of breast cancer, and thus play a growth-inhibiting action on breast cancer.

[Effect of low-dose ionizing radiation exposure on thyroid function in a medical occupational population].

Objective: We evaluated the effect of low-dose of ionizing radiation on thyroid function of medical occupational group with long-term exposure; furthermore; we analyzed the relationship between the thyroid hormones and the risk factors; such as exposure length; department. 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 were set up as occupational exposure; 724 medical professionals as the research object. We figured out the basic information and general condition of the groups by face-to-face questionnaire survey; By means of the thyroid hormone testing; we analyzed the thyroid hormone levels with different population; occupational exposure factors. Then; obtained the prevalence of thyroid nodules by the thyroid ultrasound. Besides; we used the logistic regression model to analyze the risk factors related to thyroid nodule. 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: 1. Based on the work experience; we divided the study population into four groups; such as 1-9; 10-19; 20-29; and>30 years. The difference of the TSH level among the four groups was statistically significant (P<0.05) . 2. The multiple logistic regression showed that sex and seniority were the independent risk factors for the abnormal rate of thyroid nodules. Conclusion: Long-term exposure to low-dose ionizing radiatiom could induce the thyroid damage of medical occupational population; which should be broader concerned.

Phenotypic classification of gastric signet ring cell carcinoma and its relationship with K-ras mutation.

We aimed to analyze gastric signet ring cell (SRC) carcinoma subtypes by investigating gastric and intestinal phenotypic marker expression, and explore the relationship between phenotype and K-ras mutation. Immunohistochemistry was performed on 163 SRC carcinoma patient specimens to detect gastric (MUC1, MUC5AC, and MUC6) and intestinal (MUC2 and CDX2) phenotypic markers, and tumors were classified into gastric (G), intestinal (I), and gastrointestinal (GI) phenotypes. DNA was extracted from the formalin-fixed, paraffin-embedded tumor samples, and K-ras mutations in codons 12, 13, and 61 were identified using polymerase chain reaction-based direct DNA sequencing. G, GI, and I phenotypes were observed in 63 (38.6%), 71 (43.5%), and 29 cases (17.8%), respectively. Expression of MUC2 was significantly associated with invasion depth and lymph node metastasis (P = 0.001 and 0.002, respectively), whereas that of CDX2 significantly corresponded to tumor size and submucosal invasion (P = 0.004 and 0.001, respectively). MUC5AC expression was inversely associated with gastric wall invasion (P = 0.001). Intestinal phenotypic marker expression was positively associated with gastric wall invasion and lymph node metastasis. K-ras mutations, all of which were in codon 12, were detected in 20 (12.27%) tumors, were significantly associated with the I phenotype, and exhibited an inverse relationship with MUC5AC and MUC6 expression. I-phenotype SRC carcinomas should be distinguished from those of the G phenotype because of their increased malignancy regarding invasion and metastasis, and higher K-ras aberration rate. The different K-ras mutation frequencies observed imply distinct genetic mechanisms in the carcinogenesis of I- and G-phenotype gastric SRC carcinomas.

Investigating the role of polymorphisms in miR-146a, -149, and -196a2 in the development of gastric cancer.

Here, we performed a case-control study to investigate the role of miR-146a, miR-149, and miR-196a2 polymorphisms in the development of gastric cancer using a hospital-based case-control design. A total of 186 gastric cancer patients and 186 control subjects were enrolled from Ren Ji Hospital between January 2012 and October 2014. MicroRNAs miR-146a, miR-149, and miR-196a2 were genotyped by polymerase chain reaction coupled with restriction fragment length polymorphism. Univariate logistic regression analysis revealed that patients with gastric cancer were more likely to be infected with Helicobacter pylori [odds ratio (OR) = 1.68, 95% confidence interval (CI) = 1.07-1.96]. Conditional multiple logistic regression analysis revealed that the TT genotype of miR-196a2 was associated with an increased risk of gastric cancer compared to the CC genotype (OR = 2.40; 95%CI = 1.26-4.61). Moreover, patients carrying both the TC and TT genotypes of miR-196a2 were correlated with an elevated risk of gastric cancer compared to those expressing the CC genotype alone (OR = 1.67, 95%CI = 1.01-2.75; P = 0.03). In conclusion, the results of our study indicated that the miR-196a2 polymorphism was associated with gastric cancer development.

MiR-335 functions as a tumor suppressor and regulates survivin expression in osteosarcoma.

OBJECTIVE: Previous studies have shown that miR-335 plays an anti-tumor role in several types of cancer. However, whether it is able to regulate the tumorigenesis of osteosarcoma (OS) has not been fully investigated. The present study was designed to study its potential role in regulating apoptosis of OS cells. MATERIALS AND METHODS: The expression of miR-335 in a total of 18 paired OS tumor tissues and adjacent non-cancerous tissues was measured by Real-time PCR, and its different expression in OS cell lines was also measured. The effect of miR-335 on apoptosis was measured by MTT assay, caspase-3 activity assay and TUNEL assay. The effect of survivin inhibition on apoptosis of OS cells was determined by MTT assay and western blot. Luciferase reporter assay and western blot were conducted to confirm the relationship between miR-335 and the 3'UTR of survivin mRNA. RESULTS: MiR-335 expression was found to be significantly downregulated in OS tumor tissues and OS cell lines. Overexpression of miR-335 led to decreased cell viability and increased apoptosis. MiR-335 directly targeted the 3'UTR of survivin mRNA and suppressed survivin gene expression, and inhibition of survivin exhibited similar effects to miR-335 overexpression. CONCLUSIONS: MiR-335 might function as a tumor suppressor in OS, and downregulation of miR-335 in OS cells contributes to the decreased apoptotic potential of OS cells through derepression of survivin.

Maternal molecular hydrogen treatment attenuates lipopolysaccharide-induced rat fetal lung injury.

Maternal inflammation is associated with spontaneous preterm birth and respiratory impairment among premature infants. Recently, molecular hydrogen (H2) has been reported to have a suppressive effect on oxidative stress and inflammation. The aim of this study was to evaluate the effects of H2 on fetal lung injury caused by maternal inflammation. Cell viability and the production of interleukin-6 (IL-6) and reactive oxygen species (ROS) were examined by treatment with lipopolysaccharide (LPS) contained in ordinal or H2-rich medium (HM) using a human lung epithelial cell line, A549. Pregnant Sprague Dawley rats were divided into three groups: Control, LPS, and HW + LPS groups. Rats were injected with phosphate-buffered saline (Control) or LPS intraperitoneally (LPS) on gestational day 19 and provided H2 water (HW) ad libitum for 24 h before LPS injection (HW + LPS). Fetal lung samples were collected on day 20, and the levels of apoptosis, oxidative damage, IL-6, and vascular endothelial growth factor (VEGF) were evaluated using immunohistochemistry. The number of apoptotic cells, and levels of ROS and IL-6 were significantly increased by LPS treatment, and repressed following cultured with HM in A549 cells. In the rat models, the population positive for cleaved caspase-3, 8-hydroxy-2'-deoxyguanosine, IL-6, and VEGF was significantly increased in the LPS group compared with that observed in the Control group and significantly decreased in the HW + LPS group. In this study, LPS administration induced apoptosis and oxidative damage in fetal lung cells that was ameliorated by maternal H2 intake. Antenatal H2 administration may decrease the pulmonary mobility associated with inflammation in premature infants.

A systematic review of the therapeutic interventions for heterotopic ossification after spinal cord injury.

STUDY DESIGN: Systematic review. OBJECTIVE: To conduct a systematic review of the effectiveness of interventions used to prevent and treat heterotopic ossification (HO) after spinal cord injury (SCI). SETTING: St Joseph's Parkwood Hospital, London, Ontario, Canada. METHODS: MEDLINE, CINAHL, EMBASE and PsycINFO databases were searched for articles addressing the treatment of HO after SCI. Studies were selected by two reviewers and were only included for analysis if at least 50% of the subjects had an SCI, there were at least three SCI subjects and if the study subjects participated in a treatment or intervention. Study quality was assessed by two independent reviewers using the Downs and Black evaluation tool for all studies, as well as the PEDro assessment scale for randomized control trials only. Levels of evidence were assigned using a modified Sackett scale. RESULTS: A total of 13 studies met the inclusion criteria. The selected articles were divided into prevention or treatment of post-SCI HO. Nonsteroidal anti-inflammatory drugs (NSAIDs), warfarin, and pulse low-intensity electrogmagnetic field (PLIMF) therapy were reviewed as prophylactic measures. Bisphosphonates, radiotherapy and excision were reviewed as treatments of post-SCI HO. CONCLUSIONS: Pharmacological treatments of HO after SCI had the highest level of research evidence supporting their use. Of these, NSAIDs showed greatest efficacy in the prevention of HO when administered early after an SCI, whereas bisphosphonates were the intervention with strongest supportive evidence once HO had developed. Of the non-pharmacological interventions, PLIMF was supported by the highest level of evidence; however, more research is needed to fully understand its role.

A multicenter, prospective, randomized clinical trial comparing tension-free vaginal tape surgery and no treatment for the management of stress urinary incontinence in elderly women.

AIMS: The aim of our study was to test the hypothesis that elderly women undergoing tension-free vaginal tape surgery (TVT) will have a better quality of life (QOL) and satisfaction compared to non-treated women despite age- and technique-related potential morbidity. METHODS: This multicenter, prospective, randomized, controlled trial enrolled a total of 69 women aged over 70 years who initially consented to be randomized to either undergo immediate TVT surgery or to wait for 6 months before submitting to the same surgery (control group). The main outcomes measured at every visit (pre-randomization, 8-12 weeks and 6 months) consisted of the Incontinence-Quality of Life (I-QOL) Questionnaire, the Patient Satisfaction Questionnaire and the Urinary Problems Self-assessment Questionnaire, among others. RESULTS: The analysis included 31 patients in the immediate surgery group and 27 subjects in the control group. Peri-operative complications in the immediate surgery group were bladder perforation (22.6%), urinary retention (12.9%), urinary tract infection (3.2%) and de novo urgency (3.2%). At 6 months, the mean I-QOL scores for the TVT and control groups were respectively 96.5 +/- 15.5 and 61.6 +/- 19.8 (P < 0.0001); mean Patient Satisfaction scores were respectively 8.0 +/- 2.7 and 2.0 +/- 2.4 (P < 0.0001); and mean Urinary Problems scores were respectively 4.5 +/- 4.3 and 11.6 +/- 3.5 (P < 0.0001). CONCLUSION: At 6 months post-randomization, the group of elderly women who underwent immediate TVT surgery showed a significant improvement in QOL, patient satisfaction and less urinary problems compared to the group of women waiting for the same surgery.

A rich lymphatic network exists in the inferior surface of the vocal cord.

Conservation laryngeal surgery is an increasingly available alternative for the treatment of laryngeal cancer. Understanding anatomy of laryngeal lymph drainage is essential for clinicians to diagnose, grade and surgically manage the laryngeal cancer. Although the lymphatic drainage of the larynx has been extensively studied, few studies revealed the relationship of the lymphatic drainage between various parts of the larynx. The distribution of lymphatic vessels in the inferior surface of the vocal cord also remains unclear. The aim of this study was to investigate the communication of the lymphatic networks between the vocal cord, the supraglottic and subglottic parts of the larynx, paying special attention to the lymphatic drainage of the inferior surface of the vocal cord. Eighteen larynx specimens from 18 fresh fetal cadavers were manually injected with prassion blue solution into the mucosal or submucosal layer of the larynx in order to reveal the lymphatic vessels in the inner larynx. We found that a rich lymphatic network is present in the inferior surface of the vocal cord, and the lymphatic networks in the superior and inferior surfaces of the vocal cord appear as two different patterns. These findings provide an anatomical basis for the design of a partial or conservation laryngeal surgery, particularly when considering the precise resection margin.

Structural organization of the human MS4A gene cluster on Chromosome 11q12.

CD20, the high-affinity IgE receptor beta chain (FcepsilonRIbeta), and HTm4 are structurally related cell surface proteins expressed by hematopoietic cells. Recently, 16 novel human and mouse genes were identified that encode new members of this nascent protein family that we have named the membrane-spanning 4A gene family, with at least 12 subgroups (MS4A1-MS4A12). In the current study, we identified three additional human MS4A genes: MS4A4E, MS4A6E, and MS4A10. All family members have at least four potential transmembrane domains and N- and C-terminal cytoplasmic domains encoded by distinct exons, except MS4A6E which contains two transmembrane domains. Otherwise, the 12 currently identified MS4A genes share common structural features and similar intron/exon splice boundaries, and are clustered along an approximately 600-kb region of Chromosome 11q12. In contrast to other MS4A genes, MS4A4E, MS4A6E, and MS4A10 transcripts were rare and not detected among hematopoietic cells and most nonlymphoid tissues. Sequence polymorphisms were identified in the MS4A6E gene and common splice variants were observed for the MS4A4A, MS4A5, MS4A6A, and MS4A7 genes. Thus, the MS4A family currently includes 24 distinct human and mouse genes. Like CD20 and FcepsilonRIbeta, the 10 other human MS4A family members are likely to be components of oligomeric cell surface complexes involved in signal transduction in diverse cell lineages.