ObjectiveTo investigate the effects of ethoxysanguinarine （ETH） on angiotensin Ⅱ （Ang Ⅱ）-mediated cardiomyocyte apoptosis and its regulatory effects of inositol-requiring enzyme 1 （IRE1）/regulated IRE1-dependent decay （RIDD） signaling pathway and endoplasmic reticulum stress. MethodsWestern blot was used to detect the establishment of the H9c2 model via Ang Ⅱ stimulation， which was identified as a cardiomyocyte apoptosis model. Subsequently， the inhibitory effect of ETH on cell proliferation was assessed using the cell counting Kit-8 （CCK-8） to determine the optimal effective dose of ETH. H9c2 cardiomyocytes were divided into a blank group， a model group （Ang Ⅱ， 1 mmol·L^-1）， and low-， medium-， and high-dose ETH groups （1.25， 2.5， and 5 mmol·L^-1）. Morphological changes in cardiomyocytes induced by Ang Ⅱ were detected using phalloidin staining. Cardiomyocyte apoptosis was assessed using terminal deoxynucleotidyl transferase dUTP nick and labeling （TUNEL） staining. The apoptosis cycle was detected by Annexin V/PI flow cytometry. Western blot was used to detect the expression levels of apoptosis-related proteins， endoplasmic reticulum stress， and IRE1/RIDD pathway-related proteins. ResultsWestern blot results showed that 1 mmol/mL Ang Ⅱ stimulation significantly increased the protein expression levels of Bip， p-IRE1， and Bid in H9c2 cells （P<0.05， P<0.01）， indicating the induction of endoplasmic reticulum stress， activation of the IRE1/RIDD signaling pathway， and initiation of the apoptosis process. Compared with the blank group， the model group showed a significant increase in the surface area of H9c2 cells and the apoptosis rate of cardiomyocytes， as well as in both early and late apoptosis rates （P<0.01）. The expression levels of Bid， Bax， cleaved-Caspase-3， and cleaved-Caspase-8 proteins were significantly increased， while the expression level of Bcl-2 protein was significantly decreased （P<0.01）. The expression levels of Bip， p-IRE1， and p-RIDD proteins were significantly increased （P<0.05， P<0.01）. Compared with those in the model group， the surface area of cardiomyocytes and the apoptosis rate of cardiomyocytes in all ETH groups were significantly decreased after drug intervention. Both early and late apoptosis rates were significantly decreased. The expression level of cleaved-Caspase-8 was significantly decreased in the low-dose ETH group （P<0.05）. The expression levels of Bid， Bax， and cleaved-Caspase-8 were significantly decreased in the medium-dose ETH group （P<0.05， P<0.01）. The high-dose ETH group significantly reduced the expression levels of Bid， Bax， cleaved-Caspase-3， and cleaved-Caspase-8 （P<0.05， P<0.01） and significantly increased the expression level of Bcl-2 （P<0.05）. The level of p-IRE1 protein in the medium-dose ETH group was significantly decreased （P<0.01）. The expression levels of Bip， p-IRE1， and p-RIDD proteins in the high-dose ETH group were significantly decreased （P<0.05， P<0.01）. ConclusionETH can alleviate Ang Ⅱ-mediated cardiomyocyte apoptosis by inhibiting the IRE1/RIDD signaling pathway and further alleviate the cardiac injury caused by hypertension.

ObjectiveTo investigate the effects of ethoxysanguinarine （ETH） on angiotensin Ⅱ （Ang Ⅱ）-mediated cardiomyocyte apoptosis and its regulatory effects of inositol-requiring enzyme 1 （IRE1）/regulated IRE1-dependent decay （RIDD） signaling pathway and endoplasmic reticulum stress. MethodsWestern blot was used to detect the establishment of the H9c2 model via Ang Ⅱ stimulation， which was identified as a cardiomyocyte apoptosis model. Subsequently， the inhibitory effect of ETH on cell proliferation was assessed using the cell counting Kit-8 （CCK-8） to determine the optimal effective dose of ETH. H9c2 cardiomyocytes were divided into a blank group， a model group （Ang Ⅱ， 1 mmol·L^-1）， and low-， medium-， and high-dose ETH groups （1.25， 2.5， and 5 mmol·L^-1）. Morphological changes in cardiomyocytes induced by Ang Ⅱ were detected using phalloidin staining. Cardiomyocyte apoptosis was assessed using terminal deoxynucleotidyl transferase dUTP nick and labeling （TUNEL） staining. The apoptosis cycle was detected by Annexin V/PI flow cytometry. Western blot was used to detect the expression levels of apoptosis-related proteins， endoplasmic reticulum stress， and IRE1/RIDD pathway-related proteins. ResultsWestern blot results showed that 1 mmol/mL Ang Ⅱ stimulation significantly increased the protein expression levels of Bip， p-IRE1， and Bid in H9c2 cells （P<0.05， P<0.01）， indicating the induction of endoplasmic reticulum stress， activation of the IRE1/RIDD signaling pathway， and initiation of the apoptosis process. Compared with the blank group， the model group showed a significant increase in the surface area of H9c2 cells and the apoptosis rate of cardiomyocytes， as well as in both early and late apoptosis rates （P<0.01）. The expression levels of Bid， Bax， cleaved-Caspase-3， and cleaved-Caspase-8 proteins were significantly increased， while the expression level of Bcl-2 protein was significantly decreased （P<0.01）. The expression levels of Bip， p-IRE1， and p-RIDD proteins were significantly increased （P<0.05， P<0.01）. Compared with those in the model group， the surface area of cardiomyocytes and the apoptosis rate of cardiomyocytes in all ETH groups were significantly decreased after drug intervention. Both early and late apoptosis rates were significantly decreased. The expression level of cleaved-Caspase-8 was significantly decreased in the low-dose ETH group （P<0.05）. The expression levels of Bid， Bax， and cleaved-Caspase-8 were significantly decreased in the medium-dose ETH group （P<0.05， P<0.01）. The high-dose ETH group significantly reduced the expression levels of Bid， Bax， cleaved-Caspase-3， and cleaved-Caspase-8 （P<0.05， P<0.01） and significantly increased the expression level of Bcl-2 （P<0.05）. The level of p-IRE1 protein in the medium-dose ETH group was significantly decreased （P<0.01）. The expression levels of Bip， p-IRE1， and p-RIDD proteins in the high-dose ETH group were significantly decreased （P<0.05， P<0.01）. ConclusionETH can alleviate Ang Ⅱ-mediated cardiomyocyte apoptosis by inhibiting the IRE1/RIDD signaling pathway and further alleviate the cardiac injury caused by hypertension.

Objective:To investigate the potential mechanism of miR-1298-5p in non-small cell lung cancer(NSCLC)to regu-late the MSH2 gene and its effect on the biological behavior of tumor cells and the tumor immune microenvironment.Methods:Bioin-formatics was used to identify the key genes and miRNA involved in NSCLC.Cell proliferation was detected by CCK-8 assay,and cell invasion and migration were detected by Transwell assay.Levels of inflammatory factors were detected by ELISA assay.Western blot was used to measure the expression of MSH2 in cells,and fluorescence quantitative polymerase chain reaction(RT-qPCR)was used to detect the expressions of miR-1298-5p and MSH2 gene in NSCLC cells.Dual luciferase reporter gene assay was performed to verify the targeting relationship between miR-1298-5p and MSH2.Spearman correlation analysis was performed to correlate miR-1298-5p with immune cells and immune factors in the tumor immune microenvironment.Results:The level of miR-1298-5p was down-regulated in NSCLC cells compared with normal lung tissue cells.miR-1298-5p overexpression inhibited the proliferation,migration and inva-sion of NSCLC cells.MSH2 was confirmed to be a target gene of miR-1298-5p using luciferase reporter gene assay.Furthermore,down-regulation of miR-1298-5p in NSCLC cells could be reversed by silencing MSH2.miR-1298-5p expression levels were negatively corre-lated with the levels of Treg,IL-10,and TGF-β,and positively correlated with the levels of CD3+T,CD4+T,CD8+T,NK cells,IL-2,and IFN-γ.Conclusion:miR-1298-5p negatively regulates MSH2 to inhibit the proliferation,invasion and migration of NSCLC cells and improve the tumor immune microenvironment.

Objective To construct a diagnostic model based on the information of the TCM four diagnoses in hyperactive liver yang syndrome in patients with essential hypertension.Methods Syndromic investigation was carried out in patients with essential hypertension in some hospitals in Fujian and Beijing,and diagnostic information such as age,gender,symptoms,tongue and pulse were collected.On the basis of statistical analysis,this study adopted C5.0,CRT,CHAID and QUEST decision tree models respectively.After evaluating the stability and performance consistency of the models,the accuracy of the models was measured by diagnostic rate,and the optimal model of hyperactivity of liver yang in essential hypertension was selected.Results Totally 533 patients with essential hypertension were included,including 198 patients with hyperactive liver yang syndrome and 335 patients without hyperactive liver yang syndrome.The diagnostic rates of the four models were 75.2%,66.2%,67.7%and 65.0%,respectively.The diagnostic efficiency of C5.0 was better than that of CRT,CHAID and QUEST models."Aggravation after emotional excitement,poor complexion,string-like pulse,irritability,head swelling pain,bitter mouth,heavy pulse,fatigue,dark tongue,irritability,dizziness,thin pulse,yellow fur"could be regarded as the specific items of the syndrome model of hyperactive liver yang in essential hypertension.Conclusion The C5.0 decision tree model can clearly and intuitively identify the hyperactive liver yang syndrome in essential hypertension patients based on clinical TCM four diagnostic information data,and summarize diagnostic rules.

Objective:To validate the feasibility of the gamma analysis method in the study of prescription dose conversion between logistic nanodosimetry model (LNDM) and microdosimetric kinetic model (MKM) basing on the Chinese self-developed model LNDM by applying clinical experiences of National Institute of Radiological Science (NIRS).Methods:Physical dose distributions derived from the MKM- and LNDM-based carbon ion treatment plans were compared via the method of gamma analysis under the open-source treatment planning platform matRad. In this way, the prescribed dose conversion factor between the MKM- and LNDM-based treatment plans was obtained. Using water phantoms, the influence of geometric shape, size, depth of target volume (TV), prescribed dose and field setting on the conversion factor was investigated comprehensively. Moreover, preliminary verification of the acquired conversion factor was conducted on the C-shape model and a case of liver cancer patient.Results:The conversion factor depended on the field setting rather than the TV shape. Under the condition of single field, the conversion factor was positively correlated with the size and depth of TV, and the prescribed dose. Moreover, the conversion factor was successfully verified using the C-shape model and the patient with liver cancer, where the gamma passing rates (2%/2 mm) of the physical dose distribution generated by the MKM and LNDM treatment plans were 92.79% and 91.19%, respectively.Conclusions:The conversion factors (f=D LNDM/D MKM) obtained in this study might provide guidance for the prescribed dose setting during the carbon ion treatment planning based on the LNDM. Besides, the gamma analysis method could be used for the study of the prescribed dose conversion between different models.

Objective:To explore the effects and mechanism of annexin A1 ( ANXA1)-overexpressing human adipose-derived mesenchymal stem cells (AMSCs) in the treatment of mice with acute respiratory distress syndrome (ARDS). Methods:The experimental study method was adopted. After the adult AMSCs were identified by flow cytometry, the 3 rd passage cells were selected for the follow-up experiments. According to the random number table (the same grouping method below), the cells were divided into ANXA1-overexpressing group transfected with plasmid containing RNA sequences of ANXA1 gene and no-load control group transfected with the corresponding no-load plasmid. The other cells were divided into ANXA1-knockdown group transfected with plasmid containing small interfering RNA sequences of ANXA1 gene and no-load control group transfected with the corresponding no-load plasmid. At post transfection hour (PTH) 72, the fluorescence expression was observed under a fluorescence microscope imaging system, and the protein and mRNA expressions of ANXA1 were detected by Western blotting and real-time fluorescence quantitative reverse transcription polymerase chain reaction respectively (with the sample numbers being 3). Fifty male C57BL/6J mice aged 6-8 weeks were divided into sham injury group, ARDS alone group, normal cell group, ANXA1-overexpressing group, and ANXA1-knockdown group, with 10 mice in each group. Mice in the last 4 groups were treated with endotoxin/lipopolysaccharide to make ARDS lung injury model, and mice in sham injury group were simulated to cause false injury. Immediately after injury, mice in sham injury group and ARDS alone group were injected with normal saline through the tail vein, while mice in normal cell group, ANXA1-overexpressing group, and ANXA1-knockdown group were injected with normal AMSCs, ANXA1-overexpressing AMSCs, and ANXA1-knockdown AMSCs, correspondingly. At post injection hour (PIH) 24, 5 mice in each group were selected, the Evans blue staining was performed to observe the gross staining of the right lung tissue, and the absorbance value of bronchoalveolar lavage fluid (BALF) supernatant of left lung was detected by microplate reader to evaluate the pulmonary vascular permeability. Three days after injection, the remaining 5 mice in each group were taken, the right lung tissue was collected for hematoxylin-eosin staining to observe the pathological changes and immunohistochemical staining to observe the CD11b and F4/80 positive macrophages, and the levels of tumor necrosis factor α (TNF-α), interleukin-6 (IL-6), and IL-1β in BALF supernatant of left lung were determined by enzyme-linked immunosorbent assay. Data were statistically analyzed with paired sample t test, one-way analysis of variance, and least significant difference test. Results:At PTH 72, AMSCs in both ANXA1-overexpressing group and ANXA1-knockdown group expressed higher fluorescence intensity than AMSCs in corresponding no-load control group, respectively. At PTH 72, compared with those in corresponding no-load control group, the protein and mRNA expressions of ANXA1 in ANXA1-overexpressing group were significantly increased (wth t values of 249.80 and 6.56, respectively, P<0.05), while the protein and mRNA expressions of ANXA1 in ANXA1-knockdown group were significantly decreased (wth t values of 176.50 and 18.18, respectively, P<0.05). At PIH 24, compared with those in sham injury group (with the absorbance value of BALF supernatant being 0.041±0.009), the lung tissue of mice in ARDS alone group was obviously blue-stained and the absorbance value of BALF supernatant (0.126±0.022) was significantly increased ( P<0.05). Compared with those in ARDS alone group, the degree of blue-staining in lung tissue of mice was significantly reduced in normal cell group or ANXA1-overexpressing group, and the absorbance values of BALF supernatant (0.095±0.020 and 0.069±0.015) were significantly decreased ( P<0.05), but the degree of blue-staining in lung tissue and the absorbance value of BALF supernatant (0.109±0.016, P>0.05) of mice in ANXA1-knockdown group had no significant change. Compared with that in normal cell group, the absorbance value of BALF supernatant of mice in ANXA1-overexpressing group was significantly decreased ( P<0.05). Three days after injection, the lung tissue structure of mice in ARDS alone group was significantly damaged compared with that in sham injury group. Compared with those in ARDS alone group, hemorrhage, infiltration of inflammatory cells, alveolar collapse, and interstitial widening in the lung tissue of mice were significantly alleviated in normal cell group and ANXA1-overexpressing group, while no significant improvement of above-mentioned lung tissue manifestation was observed in ANXA1-knockdown group. Three days after injection, the numbers of CD11b and F4/80 positive macrophages in the lung tissue of mice in ARDS alone group were significantly increased compared with those in sham injury group. Compared with those in ARDS alone group, the numbers of CD11b and F4/80 positive macrophages in lung tissue of mice in normal cell group, ANXA1-overexpressing group, and ANXA1-knockdown group reduced, with the most significant reduction in ANXA1-overexpressing group. Three days after injection, compared with those in sham injury group, the levels of TNF-α, IL-6, and IL-1β in BALF supernatant of mice in ARDS alone group were significantly increased ( P<0.05). Compared with those in ARDS alone group, the levels of TNF-α, IL-6, and IL-1β in BALF supernatant of mice in normal cell group and ANXA1-overexpressing group, as well as the level of IL-1β in BALF supernatant of mice in ANXA1-knockdown group were significantly decreased ( P<0.05). Compared with that in normal cell group, the level of TNF-α in BALF supernatant of mice was significantly decreased in ANXA1-overexpressing group ( P<0.05) but significantly increased in ANXA1-knockdown group ( P<0.05). Conclusions:Overexpression of ANXA1 can optimize the efficacy of AMSCs in treating ARDS and enhance the effects of these cells in inhibiting inflammatory response and improving pulmonary vascular permeability, thereby alleviating lung injury of mice with ARDS.

Vitamin D deficiency can be a trigger factor for recurrent oral ulcers. This article combining two cases of recurrent oral ulcers in patients with vitamin D deficiency who were cured by active vitamin D supplementation emphasizes that 25-hydroxyvitamin D and other indicators should be screened in patients with recurrent oral ulcers, so as to improve the understanding of the disease and the ability of clinical diagnosis and treatment for it.

With the development of various laboratory testing techniques such as serum calcium and parathyroid hormone (PTH) , a large number of asymptomatic or normocalcemic primary hyperparathyroidism (PHPT) can be diagnosed early. PHPT has become the third most common endocrine disease affecting human health. Currently, most PHPT, especially normocalcemic primary hyperparathyroidism, are not primary diseases and may be related to vitamin D deficiency/insufficiency and/or insufficient calcium supplementation. That is, the relative hypocalcemia caused by long-term vitamin D deficiency/insufficiency and/or insufficient calcium supplementation leads to parathyroid hyperfunction, stimulates parathyroid hyperplasia, and secretes excessive parathyroid hormone to compensate for the regulation of calcium and phosphorus balance. When it is in the initial reversible stage, it can be cured by internal medicine; if it progresses freely, long-term hypocalcemia stimulation will lead to excessive parathyroid hyperplasia and even tumor occurrence with the formation of so-called PHPT and parathyroidectomy has to be performed. Therefore, routine screening of bone mineral density, calcium, magnesium, phosphours, 25-hydroxyvitamin D, parathyroid hormone and other bone metabolism indicators in the physical examination of general population is beneficial to the prevention and treatment of bone metabolism diseases, urinary stones and hyperparathyroidism. At the same time, attention should be paid to identifying the stage of prehyperparathyroidism in which vitamin D deficiency/insufficiency and insufficient calcium supplementation will stimulate parathyroid hyperfunction. Active intervention on prehyperparathyroidism is an effective way to avoid the development of primary hyperparathyroidism.

Objective:To compare the population characteristics, the positive rate of screening, the detection rate of breast cancer, early diagnosis rate and the cost between the mass screening group and opportunistic screening group of breast cancer.Methods:This study is a prospective multicenter cohort study conducted from January 1, 2014 to December 31, 2016. The participants were enrolled for mass screening or opportunistic screening of breast cancer. After completing the questionnaire, all the participants received breast physical examination and breast ultrasound examination every year for 3 rounds by year. The participants′ characteristics and screening results of the two groups were compared by χ 2 test, Fisher exact test or Wilcoxon rank-sum test. Results:A total of 20 080 subjects were enrolled. In the mass screening group, 9 434 (100%), 8 111 (85.98%) and 3 940 (41.76%) cases completed the 3 rounds of screening, and 10 646 (100%), 6 209 (58.32%) and 2 988 (28.07%) cases in the opportunistic screening group, respectively. In the opportunistic screening group, the proportions of less than 3 months lactation (1 275/9 796 vs. 1 061/8 860, χ2=4.597, P=0.032), non-fertility (850/10 646 vs. 574/9 434, χ2=27.400, P<0.01), abortion history (6 384/10 646 vs. 5 062/9 434, χ2=81.232, P<0.01), postmenopausal (2 776/10 646 vs. 2 217/9 434, χ2=17.757, P<0.01), long-term oral contraceptives(>6 months) (171/10 646 vs. 77/9 434, χ2=25.593, P<0.01) and family history of breast cancer in first-degree relatives (464/10 646 vs. 236/9 434, χ2=51.257, P<0.01) were significantly higher than those in mass screening group. The positive rate of screening (514/10 646 vs. 128/9 434, χ2=194.736, P<0.01), the detection rate of breast cancer (158/10 646 vs. 13/9 434, χ2=107.374, P<0.01), and positive rate of biopsy (158/452 vs. 13/87, χ2=13.491, P<0.01) in the opportunistic screening group were significantly higher than those of the mass screening group. The early diagnosis rate of the mass screening group was significantly higher than the opportunistic screening group (10/12 vs. 66/141, χ2=5.902, P=0.015). The average cost for detecting each breast cancer case of the mass screening group was 215 038 CNY, which was 13.6 times of the opportunistic screening group (15 799 CNY/case). In the opportunistic screening group, the positive rate of biopsy in primary hospitals was significantly lower than that in large-volume hospitals (79/267 vs. 79/185, χ2=8.267, P=0.004), but there was no significant difference in the mass screening group (6/37 vs. 7/50, χ2=0.082, P=0.774). Conclusions:Breast cancer screening can improve early detection rate. Compared with the mass screening mode, the opportunistic screening mode has the advantages of higher proportion of high-risk factors, higher positive rate of screening, higher detection rate of breast cancer, higher positive rate of biopsy, and lower cost of screening. However, the early diagnosis rate of breast cancer of opportunistic screening is lower than that of mass screening. The positive rate of opportunistic screening in primary hospitals is lower than that of large-volume hospitals. The two screening modes have their own advantages and should be chosen according to local conditions of different regions in China.

Objective:To compare the population characteristics, the positive rate of screening, the detection rate of breast cancer, early diagnosis rate and the cost between the mass screening group and opportunistic screening group of breast cancer.Methods:This study is a prospective multicenter cohort study conducted from January 1, 2014 to December 31, 2016. The participants were enrolled for mass screening or opportunistic screening of breast cancer. After completing the questionnaire, all the participants received breast physical examination and breast ultrasound examination every year for 3 rounds by year. The participants′ characteristics and screening results of the two groups were compared by χ 2 test, Fisher exact test or Wilcoxon rank-sum test. Results:A total of 20 080 subjects were enrolled. In the mass screening group, 9 434 (100%), 8 111 (85.98%) and 3 940 (41.76%) cases completed the 3 rounds of screening, and 10 646 (100%), 6 209 (58.32%) and 2 988 (28.07%) cases in the opportunistic screening group, respectively. In the opportunistic screening group, the proportions of less than 3 months lactation (1 275/9 796 vs. 1 061/8 860, χ2=4.597, P=0.032), non-fertility (850/10 646 vs. 574/9 434, χ2=27.400, P<0.01), abortion history (6 384/10 646 vs. 5 062/9 434, χ2=81.232, P<0.01), postmenopausal (2 776/10 646 vs. 2 217/9 434, χ2=17.757, P<0.01), long-term oral contraceptives(>6 months) (171/10 646 vs. 77/9 434, χ2=25.593, P<0.01) and family history of breast cancer in first-degree relatives (464/10 646 vs. 236/9 434, χ2=51.257, P<0.01) were significantly higher than those in mass screening group. The positive rate of screening (514/10 646 vs. 128/9 434, χ2=194.736, P<0.01), the detection rate of breast cancer (158/10 646 vs. 13/9 434, χ2=107.374, P<0.01), and positive rate of biopsy (158/452 vs. 13/87, χ2=13.491, P<0.01) in the opportunistic screening group were significantly higher than those of the mass screening group. The early diagnosis rate of the mass screening group was significantly higher than the opportunistic screening group (10/12 vs. 66/141, χ2=5.902, P=0.015). The average cost for detecting each breast cancer case of the mass screening group was 215 038 CNY, which was 13.6 times of the opportunistic screening group (15 799 CNY/case). In the opportunistic screening group, the positive rate of biopsy in primary hospitals was significantly lower than that in large-volume hospitals (79/267 vs. 79/185, χ2=8.267, P=0.004), but there was no significant difference in the mass screening group (6/37 vs. 7/50, χ2=0.082, P=0.774). Conclusions:Breast cancer screening can improve early detection rate. Compared with the mass screening mode, the opportunistic screening mode has the advantages of higher proportion of high-risk factors, higher positive rate of screening, higher detection rate of breast cancer, higher positive rate of biopsy, and lower cost of screening. However, the early diagnosis rate of breast cancer of opportunistic screening is lower than that of mass screening. The positive rate of opportunistic screening in primary hospitals is lower than that of large-volume hospitals. The two screening modes have their own advantages and should be chosen according to local conditions of different regions in China.