[Bronchoscopic manifestations and interventional treatment of pulmonary mucormycosis].

Objective: To analyze the bronchoscopic manifestations and interventional treatment of pulmonary mucormycosis. Methods: Clinical data of patients with pulmonary mucormycosis undergoing bronchoscopy and interventional therapy in 4 tertiary general hospitals in China from May 2006 to May 2022 were retrospectively analyzed and the literature on the subject were reviewed. Results: The data of 10 patients with pathologically diagnosed pulmonary mucormycosis undergoing bronchoscopy and interventional therapy were collected, including 8 males and 2 females. The patients' age ranged from 21 to 72 (44±15) years. The underlying diseases included 6 cases of diabetes ketoacidosis, 3 cases of leukemia, 1 case after operation of lung cancer. Bronchoscopy showed that white viscous necrotic matters grew along the airway and blocked the airway in 9 cases, accompanied by airway bleeding in 3 cases, bloody secretion blocked the airway in 1 case, and bronchopulmonary cavity fistula in 2 cases. The biopsy histopathology of white necrotic matters showed that many mucor filaments were tangled together which were named mucormycelium. Among the 10 patients, 9 were treated with systemic drugs, including intravenous application of amphotericin B deoxycholate in 5 cases, intravenous application of amphotericin B liposome in 4 cases, oral posaconazole in 6 cases and intravenous injection in 1 case. Local drug therapy included aerosol inhalation of amphotericin B deoxycholate in 8 cases and local perfusion under bronchoscope in 5 cases. Bronchoscopic interventional therapy was used to remove mucormycelium in the bronchus, including cryotherapy in 8 cases, biopsy forceps in 7 cases, snare treatment in 2 cases and foreign body forceps in 2 cases. All 10 patients were clinical cured and with no death. Conclusions: Pulmonary mucormycosis is more common in immunocompromised hosts. Bronchoscopy often showed mucormycelium blocking the airway. Systemic and local drug therapy combined with bronchoscopic interventional therapy can achieve good clinical efficacy.

[Study on mechanisms of Th17/Treg imbalance in patients with cystic echinococcosis based on miRNA expression profiles].

OBJECTIVE: To investigate the serum microRNA (miRNA) expression and examine the impact of miRNA expression profiles on T helper type 17 (Th17)/regulatory T cells (Treg) imbalance among patients with cystic echinococcosis, so as to provide insights into the illustration of the mechanisms underlying chronic Echinococcus granulosus infections, and long-term pathogenesis. METHODS: Total RNA was extracted from the sera of cystic echinococcosis patients and healthy controls, and subjected to high-throughput sequencing with the Illumina sequencing platform. Known miRNAs were annotated and new miRNAs were predicted using the miRBase database and the miRDeep2 tool, and differentially expressed miRNAs were identified. The target genes of differentially expressed miRNAs were predicted using the software miRanda and TargetScan, and the intersection was selected for Gene Ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. Among the differentially expressed miRNAs with the 20 highest fold changes, miRNAs that targeted genes relating to key transcription factors RORC and FOXP3 that determine the production of Th17 and Treg cells or their important regulatory pathways (PI3K-Akt and mTOR pathways) were matched. RESULTS: A total of 53 differentially expressed miRNAs were screened in sera of cystic echinococcosis patients and healthy controls, including 47 up-regulated miRNAs and 6 down-regulated miRNAs. GO enrichment analysis showed that these differentially expressed miRNA were involved DNA transcription and translation, cell components, cell morphology, neurodevelopment and metabolic decomposition, and KEGG pathway analysis showed that the differentially expressed miRNA were mainly involved in MAPK, PI3K-Akt and mTOR signaling pathways. Among the differentially expressed miRNAs with the 20 highest fold changes, there were 3 miRNAs that had a potential for target regulation of RORC, and 15 miRNAs that had a potential to target the PI3K-Akt and mTOR signaling pathways. CONCLUSIONS: Significant changes are found in serum miRNA expression profiles among patients with E. granulosus infections, and differentially expressed miRNAs may lead to Th17/Treg imbalance through targeting the key transcription factors of Th17/Treg or PI3K-Akt and mTOR pathways, which facilitates the long-term parasitism of E. granulosus in hosts and causes a chronic disease.

[The effect of occupational stress on anxiety of nursing staff in a third-grade general hospital and mediating effect of job burnout].

Objective: To explore the mediating effect of job burnout of nursing staff in clinical departments on occupational stress and anxiety, and to provide scientific basis for the formulation of intervention measures to relieve anxiety. Methods: From November 2020 to January 2021, a cross-sectional survey was conducted to investigate the basic situation, occupational stress, job burnout and anxiety of 653 nursing staff in a third class A general hospital in Hebei Province. Spearman rank correlation was used to analyze the relationship between occupational stress, job burnout and anxiety, stepwise regression and mediating effect model were used to verify the mediating effect of job burnout on the relationship between occupational stress and anxiety. Results: 551 valid questionnaires were collected with effective recovery of 84.38%. The incidence of high occupational stress was 68.06% (375/551) , the incidence of job burnout was 63.70% (351/551) [high, moderate and moderate were 11.07% (61/551) and 52.63% (290/551) respectively], and the incidence of anxiety was 55.72% (307/551) [mild, moderate and severe were 38.11% (210/551) , 8.53% (47/551) and 9.08% (50/551) respectively]. Occupational stress was positively correlated with job burnout and anxiety (r=0.545, 0.479) , and job burnout was positively correlated with anxiety (r=0.542, P<0.05) . The mediating effect analysis showed that occupational stress had a statistically significant effect on anxiety (c=0.509, P<0.001) , and the mediating effect of job burnout on the relationship between occupational stress and anxiety accounted for 44.99% of the total effect. Conclusion: The anxiety level of the nursing staff in this third-class A general hospital was relatively high. Job burnout has a mediating effect between occupational stress and anxiety, and anxiety of nursing staff can be alleviated by reducing occupational stress or job burnout.

[Increased risk of cardiovascular disease in elderly population with carotid plaque and low ankle brachial index].

Objective: To investigate whether the co-presence of carotid plaques and low ankle-brachial index (ABI) might increase the risks of ischemic cardiovascular and cerebrovascular event in elderly population. Methods: It was a prospective study. Participants from the elderly cohort of the Kailuan Study, who completed a carotid sonography and ABI examination, were included in this study. Participants underwent physical examinations between 2010 and 2011 and were divided into 3 groups: no carotid plaque and ABI>0.9 group (n=526), carotid plaque and ABI>0.9 group (n=1 067), and carotid plaques and ABI≤0.9 group (n=49). Follow up ended on the 31 December 2016. The incidence of ischemic cardiovascular and cerebrovascular event was compared between the 3 groups, the relationship between carotid plaque and low ABI with ischemic cardiovascular and cerebrovascular event was analyzed. Results: A total of 1 642 participants were included (age, (67.1±6.4) years). There were 1 028 males (62.6%) and 1 028 females(37.4%). The average follow-up time was 5.41 years, the incidence of ischemic cardiovascular and cerebrovascular event in the 3 group was 2.1%(11/526), 5.5%(59/1 067), and 12.2%(6/49),respectively; the incidence of myocardial infarction in the 3 group was 0.2%(1/526), 1.6%(17/1 067), 10.2%(5/49), respectively; the incidence of cerebral infarction in the 3 group was 1.9%(10/526), 3.9%(42/1 067) and 2.0%(1/49), respectively. Multivariate Cox risk proportional regression analysis showed that compared with the group without carotid plaque and ABI>0.9, the HR values (95%CI) of ischemic cardiovascular and cerebrovascular event in the group with carotid plaque and ABI>0.9, carotid plaques and ABI≤0.9 group were 3.52 (1.49-8.35), 7.16(2.11-24.26) respectively, after adjusting for sex,age,systolic blood pressure,fast blood glucose,body mass index,total cholesterol,smoke,alcohol consumption and lipid-lowering medication and antihypertensive medication. Conclusions: Co-presence of carotid plaques and low ankle-brachial index may further increase the risk of ischemic cardiovascular and cerebrovascular event among elderly population in this cohort.

Zoledronic acid and fracture risk: a meta-analysis of 12 randomized controlled trials.

OBJECTIVE: Zoledronic acid is widely used in patients with osteoporosis, and this meta-analysis aims to explore the influence of zoledronic acid on fracture risk and mortality in patients with osteoporosis or osteopenia. MATERIALS AND METHODS: We searched PubMed, Google Scholar, and Cochrane Library for randomized clinical trials comparing zoledronic acid with control intervention (i.e., placebo or nothing) for osteoporosis or osteopenia. The fracture and mortality were estimated using the random-effect model. RESULTS: 12 randomized trials were included in this meta-analysis. Compared to control intervention, zoledronic acid was associated with significantly reduced incidence of fracture at the follow-up of 12 months, 24 months, 36 months and 72 months. In addition, zoledronic acid could remarkably reduce mortality at 12 months and 24 months than control intervention but revealed no influence on mortality at 36 months or 72 months. In terms of adverse events, zoledronic acid might result in the increase in serious atrial fibrillation and death from stroke than control intervention. CONCLUSIONS: Zoledronic acid is beneficial to reduce the incidence of fracture, while its benefits to reduce the mortality are only observed at the follow-up time of 24 months.

ISOC1 promotes the proliferation of gastric cancer cells by positively regulating CDK19.

OBJECTIVE: To explore the role and potential mechanism of isochorismatase domain-containing 1 (ISOC1) in gastric cancer. PATIENTS AND METHODS: The expression levels of ISOC1 in gastric cancer (GC) tissues, as well as corresponding cell lines, was evaluated by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). A cell line stably expressing ISOC1 was constructed by vector construction and cell transfection, and the proliferation ability of the stably transfected cells was examined. Subsequently, the ISOC1 target database was screened, which suggested that CDK19 may be the potential target. The correlation between ISOC1 and CDK19 mRNA and protein expressions in clinical tissue specimens and cell lines was evaluated by qRT-PCR and Western blot, and the Luciferase reporter gene experiment was applied to verify the regulatory effect of ISOC1 on CDK19. RESULTS: ISOC1 was shown to be markedly increased in GC tissues compared to adjacent cancer tissues by qRT-PCR. In addition, compared with patients with low ISOC1 expression, the pathological stage and tumor size of gastric cancer patients with high ISOC1 expression were remarkably larger. Then, the ISOC1 knockdown cell line was established, and it was found through cell proliferation function experiments that the proliferation rate of gastric cancer cells was remarkably slower than the control group after knocking down ISOC1. Subsequently, bioinformatics and Luciferase reporter gene experiments suggested that ISOC1 had a direct regulatory effect on CDK19. In addition, recovery experiments also demonstrated that CDK19 overexpression could reverse the effect of ISOC1 silencing on cell proliferation. CONCLUSIONS: ISOC1 was markedly upregulated in GC tissues. It could positively regulate its downstream target CDK19, which in turn promoted the proliferation of GC cells. Therefore, our study may provide new ideas for understanding the progression of GC.

[Effects of alprostadil in ß-aminopropanitrile induced aortic dissection in a murine model].

Objective: To investigate the protective role of alprostadil on aortic dissection. Methods: 26 C57BL6 male mice were divided into control group (normal drinking water, n=13) and model group (1 g·kg-1·d-1 BAPN via drinking water, n=13). On day 14, mRNA expression of inflammatory-related genes as well as EP receptor families were detected by RT-PCR (n=6 each) and EP4 protein levels were determined by Western blot (n=7 each). Another 88 mice were divided into 3 groups: control group (n=22), model group (n=33) and treatment group (n=33). The mice in model group and treatment group were applied with BAPN (1 g·kg-1·d-1) via drinking water. The mice in treatment group received additional intraperitoneal injection with alprostadil (80 µg·kg-1·d-1) for 28 days. The mice in the control and model group received equal volume intraperitoneal injection with 0.9% saline respectively. The body weight and systolic blood pressure, the mortality and morbidity were monitored from the beginning until the designed end of the study. On day 28, the mice were sacrificed and aorta were fixed, embedded and sliced, followed by staining with HE and Victoria Blue. The distribution of EP4 was determined by immunohistochemistry in control (n=6) and model group (n=6). Furthermore, the concentration of PGE1 were tested among model (n=3) and treatment group (n=4). EP4 protein expression was determined in model group (n=7) and treatment group (n=6). Results: On day 14, mRNA expression level of MCP-1 ((2.74±1.55) vs. (1.00±0.49),<0.05) and MMP2((1.38±0.42) vs. (1.00±0.27), P<0.05) was significantly upregulated in model group compared with control group. Protein expression of EP4 receptor also increased in aorta in model group compared with control group (1.48±0.51 vs. 1.00±0.19, P<0.05). In the dissection area, the EP4 expression was also enriched compared with non-dissection area, particularly in endothelial cells and inflammatory cells on day 28. BAPN applied in drinking water (model and treatment groups) successfully induced the aortic dissection in mice, some mice died of the rupture. The elastic fibers were fractured, and the infiltrated immune cells were visible in dissected tissue. False lumen was formed. There was no dissection and death in the control group. Compared with control group, the morbidity and mortality rates were significantly increased in the model group (60.6%, 20/33, 30.3%, 10/33) and the treatment group (72.7%, 24/33, 24.2%, 8/33). The mortality and morbidity rates were similar between model and treatment groups. There is no difference in terms of SBP among three groups (P>0.05). Further study showed that after alprostadil injection, the blood concentration of PGE1 was increased in treatment group ((0.540±0.041 vs. 0.436±0.012)µmol/L, P<0.05). Besides, the EP4 receptor expression was downregulated in the treatment group compared to model group (0.60±0.30 vs. 1.00±0.20, P<0.05). Conclusion: EP4 expression is upregulated in BAPN induced aortic dissection mouse model. No protective effects are observed post alprostadil treatment in this model probably due to the reduced expression of EP4.

[Spatial autocorrelation and its application in public health field].

The prevalence and distribution of infectious diseases and occupational diseases in specific time and space is the result of the interaction between natural environment, social economy and other factors, its distribution pattern has spatial properties. Based on the assumption of independence, the traditional statistical methods ignore the spatial attributes of diseases and cannot analyze the spatial characteristics of diseases. On the basis of geographic information system, the spatial autocorrelation analysis can simultaneously analyze the spatial relationship and attribute value of diseases, explore the spatial dependence of disease data in different spatial units, and provide decision-making basis for the prediction and early warning of diseases such as occupational and infectious diseases, and the formulation and evaluation of prevention and control measures.

Metformin reduces pancreatic cancer cell proliferation and increases apoptosis through MTOR signaling pathway and its dose-effect relationship.

OBJECTIVE: To study the influences of metformin on the proliferation and apoptosis of pancreatic cancer cells and its dose-effect relationship and crucial molecular mechanism. MATERIALS AND METHODS: With human pancreatic cancer cell line PANC-1 as the study object, different concentrations of metformin were added for intervention. Then, the proliferation of PANC-1 cells was detected via methyl thiazolyl tetrazolium (MTT) assay to determine the dose-effect relationship of metformin in PANC-1 cell proliferation. PANC-1 cells were treated with metformin at three appropriate concentrations as Metformin treatment groups, and an equal amount of dimethyl sulfoxide (DMSO) was added in Control group. Flow cytometry was performed to detect PANC-1 cell cycle and apoptosis, and the apoptosis of PANC-1 cells was also evaluated via terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay. Caspase-3 protein localization and expression in PANC-1 cells were detected using immunofluorescence assay. Besides, the expressions of the apoptosis-associated proteins Caspase-3, B-cell lymphoma 2 (Bcl-2), and Bcl-2 associated X protein (Bax) and phosphatidylinositol 3-hydroxy kinase (PI3K), phosphorylated protein kinase B (p-Akt), and p-mammalian target of rapamycin (mTOR) proteins related to the mTOR pathway were detected using Western blotting. RESULTS: Metformin repressed the proliferation of human pancreatic cancer PANC-1 cells in a concentration-dependent and time-dependent manner. Compared with Control group, Metformin treatment groups (0, 20 and 40 mM) exhibited a higher proportion of PANC-1 cells in G0/G1 phases, and a lower proportion of PANC-1 cells in S phase (p<0.05), and the change in the proportion of cells in G2/M phase was not statistically significant (p>0.05). Moreover, Metformin treatment groups (0, 20, and 40 mM) had more apoptotic PANC-1 cells, higher expression levels of pro-apoptosis proteins Caspase-3 and Bax and lower expression levels of anti-apoptosis protein Bcl-2 and the mTOR pathway-related proteins PI3K, p-Akt, and p-mTOR in cells than Control group (p<0.05). CONCLUSIONS: Metformin modulates the mTOR signaling pathway to reduce the proliferation of pancreatic cancer cell, but increase their apoptosis.

[The characteristics of spatial-temporal evolvement of pneumoconiosis in Hebei Province from 2009 to 2018].

Objective: To explore the temporal and spatial characteristics of pneumoconiosis in Hebei Province from 2009 to 2018, and to provide evidence for the policy development of pneumoconiosis prevention and control. Methods: In February 2019, a database of pneumoconiosis incidence from 2009 to 2018 in Hebei Province was built. The spatial-temporal distribution of pneumoconiosis in Hebei Province was displayed based on barycenter migration technology and spatial autocorrelation analysis. Results: A total of 6099 cases of pneumoconiosis were reported in Hebei Province from 2009 to 2018, the top 5 pneumoconiosis were silicosis(4399, 72.13%) , coal worker pneumoconiosis (1298 , 21.28%) , ceramics pneumoconiosis (224, 3.67%) , welding worker's pneumoconiosis(76, 1.25%) , and casting worker's pneumoconiosis(48, 0.79%). The focus of pneumoconiosis from 2009 to 2014 moved from Tangshan in the northeast to Zhangjiakou in the northwest, and moved to Chengde in the north from 2015 to 2017, and moved back to Tangshan in 2018. The incidence of pneumoconiosis was globally autocorrelation (P <0.05) from 2009 to 2011 and 2014 to 2018. The high-high gathering areas were located in Zhangjiakou, Chengde and other areas. Conclusion: The barycenter of pneumo- coniosis in hebei province from 2009 to 2018 is relatively stable, mainly locates in the northern part of Hebei Province over the years. There is an obvious regional aggregation, and the aggregation type is mainly high-high aggregation, which indicates that relevant departments should focus on strengthening the prevention and treat- ment of aggregation areas on the basis of overall planning.

Characteristic analysis of Zenker's diverticulum incidentally detected on multimodal neck ultrasound.

PURPOSE: Zenker's diverticulum (ZD) incidentally detected on neck ultrasound (US) could be easily misdiagnosed as a thyroid nodule and unnecessarily removed by surgical operation. It is a critical issue to identify the characteristics of ZD in clinical practice. METHODS: We reported 10 cases of ZD diagnosed using multimodal US, and discussed the features of its multimodal US images extracted from grayscale sonograms, color Doppler flow imaging (CDFI), three-dimensional US (3D-US), and contrast-enhanced US (CEUS). RESULTS: All lesions were heterogeneous in the posterior of the left thyroid. CDFI showed no blood flow within the lesions. The shapes and internal echoes of the masses changed when patients swallowed or drank. Performing CEUS or 3D-US for those with atypical US findings was complementary to make the final diagnosis. Importantly, multimodal US helped to diagnose suspected small lesions in three cases in which the barium esophagram tests proved negative. CONCLUSIONS: The US features of ZD detected by multiple modes of US can help in the collection of comprehensive imaging information on ZD, which should be used proactively to facilitate the correct diagnosis and to avoid misdiagnosis of ZD.

MiR-212-5p regulates the proliferation and apoptosis of AML cells through targeting FZD5.

OBJECTIVE: To explore the effects of microRNA-212-5p (miR-212-5p) on biological functions of acute myeloid leukemia (AML) and to find the potential molecular mechanism. PATIENTS AND METHODS: We measured the expression level of miR-212-5p in 35 AML patients and 20 patients with idiopathic thrombocytopenic purpura (ITP) as control cases. Besides, the miR-212-5p expression at cellular level was checked as well. In order to screen the functional targets of miR-212-5p, online prediction software was used and gene frizzled class receptor 5 (FZD5) attracted our attention. The effects of miR-212-5p on biological functions of AML cell line (Kasumi-1) were analyzed by subsequent experiments. The mRNA and protein expressions of FZD5 were detected by quantitative Real-time polymerase chain reaction (qRT-PCR) and Western blotting (WB) analysis, respectively. Cell proliferation was tested by cell counting kit-8 (CCK-8) assay. Cell cycle and apoptosis were measured by flow cytometry. Finally, protein expression of ß-catenin was analyzed by WB assay. RESULTS: In AML cases and cells, miR-212-5p was found to be lowly expressed. The potential target of miR-212-5p was predicated in three public databases. Through a series of experiments including qRT-PCR, WB and luciferase assay, we identified FZD5 as a functional target of miR-212-5p. In further cellular functional experiments on Kasumi-1, we found overexpression of miR-212-5p in Kasumi-1 cells greatly inhibited the cell viability and proliferation. The ratio of cells in G0/G1 phase and the proportion of apoptotic cells increased after miR-212-5p overexpression. Furthermore, Wnt/ß-catenin signal pathway was the most apparent pathway that was regulated by miR-212-5p according to WB results. However, the effects of miR-212-5p were suppressed after restoring the expression of FZD5. CONCLUSIONS: Expression of miR-212-5p was significantly lower in AML patients and cell lines, indicating that miR-212-5p served as a tumor-suppressor gene in AML. According to our in vitro experiments, miR-212-5p/FZD5 was likely to become a new therapeutic target for AML.

[Resting metabolic rate estimated by bioelectrical impedance analysis and its determinants in maintenance hemodialysis patients].

Objective: To explore the level of resting energy expenditure (REE) estimated by bioelectrical impedance analysis and the association of resting metabolic rate (RMR) with clinical related factors, and provide new ideas for improving protein energy wasting (PEW) in maintenance hemodialysis (MHD) patients. Methods: Seven hundred and sixty-five subjects receiving MHD between July 2015 and September 2016 in 11 hemodialysis centers in Guizhou province were enrolled in this cross-sectional study. Bioelectrical impedance analysis was used to measure RMR and body composition, such as lean body mass, fat mass and body cell mass (BCM). Baseline characteristics, routine blood test indexes and biochemical data of hemodialysis patients were collected. The level of RMR and body composition in hemodialysis patients was compared by gender grouping. Then the patients were divided into four groups according to the cutoff value of RMR quartile. Spearman correlation analysis and multiple linear regression analysis were used to analyze the relationships between RMR and clinical related factors. Results: The average age of MHD patients was (54.96±15.78) years and the duriation of dialysis was (42.3±9.0) months. The level of RMR in male patients (474 cases, 61.96%) was significantly higher than that in female patients [1 591(1 444, 1 764) kcal/d vs 1 226 (1 104, 1 354) kcal/d, P<0.001]. However, this significant difference of RMR between different genders disappeared after adjusting for lean body mass (P=0.193). Multiple linear regression analysis showed that RMR was positively correlated with body surface area (ß=0.817) and lactate dehydrogenase (LDH) (ß=0.198), and negatively correlated with age (ß=-0.141), all P<0.05. Conclusion: RMR levels in patients with maintenance hemodialysis are associated with lactate dehydrogenase level, which may become a new index to evaluate energy consumption.

[A review on the RET proto-oncogene mutation in medullary thyroid carcinoma].

Medullary thyroid carcinoma（MTC） is a special type of neuroendocrine tumor originated from C-cells of the thyroid gland, MTC can be divided into sporadic(70%-80%)and hereditary(20%-30%), about 98% of the hereditary MTC patients have RET proto-oncogene germline mutation in exon 10, 11, 13, 14, 15, 16. The mutation of RET proto oncogene is closely related to the pathogenesis of MTC, and different mutation of RET proto oncogene exon may lead to different MTC phenotypes.More than 100 kinds of mutations in the RET gene were reported. This paper reviews the research progress of RET proto-oncogene mutation in MTC.

Current views of depressive disorders in children - a review.

OBJECTIVE: Depression is one of the most debilitating worldwide mental illnesses, which is included in the Diagnostic and Statistical Manual of Mental Disorders. The main risk factors for the development of depression are female gender, early life stress family history of mood disorders, former anxiety disorders, substance abuse and life events. The present review article is focused on the various causative factors like childhood early life stress, cortisol or cortisol awakening response. The review would update readers with regards to the current view of factors responsible for pediatric depression, which in turn helps to plan better therapeutic approaches in the near future.

[Peripheral blood circulating tumor cells in local advanced head and neck squamous cell carcinoma].

Objective: To investigate the value of detecting circulating tumor cells (CTCs) in patients with local advanced head and neck squamous cell carcinoma (LAHNSCC). Methods: Twenty cases of LAHNSCC and eight healthy cases as the negative control were collected. The clinicopathological factors were evaluated. The LAHNSCC CTCs were enriched by specific antibody nanofluidic chip immunoassay using CytoSorter CTCs sorting system. LAHNSCC CTCs were identified by immunofluorescence staining. The relationships between CTCs and the clinicopathological features of LAHNSCC were analyzed. The numbers of CTCs were monitored and compared two weeks after inductive chemotherapy and at the end of the treatment. Results: CTCs were detect in 15 (75%) 20 patients with LASHNCC, with an average number of 22.4 CTCs. There was a correlation between the numbers of CTCs and age or N staging (P<0.05). Among the 15 cases with CTCs, 13 cases received inductive chemotherapy, for whom CTCs were detected again after inductive chemotherapy in all of these 13 patients, with an average number of 9.5 CTCs. Ten of the fourteen cases (71.4%) were still CTCs detected After whole treatments CTCs were detected in 14 patients, of them, 10 (71.4%) patients showed positive CTCs, with an average 1.6 CTCs. The numbers of CTCs decreased after either inductive chemotherapy or the whole treatment. The number of detected CTCs after whole treatment decreased nearly to background levels. Conclusions: CTCs have a high detection rate in the peripheral blood of patients with LAHNSCC, especially in patients ≥60 years old and with ≥ N2 stage before treatment. Real-time detection of dynamic change of CTCs may assist to evaluate therapeutic effect.

[Cancer-associated-fibroblasts regulate the chemoresistance of lung cancer cell line A549 via SDF-1 secretion].

Objective: To investigate whether cancer-associated- fibroblasts (CAF), the key component of tumor microenvironment, regulate the chemoresistant capacity of lung cancer cell line A549 through SDF-1 secretion. Methods: Primary cell isolation techniques was used to isolate cancer-associated-fibroblasts from lung cancer patients. MTT assay was applied to determine the proliferation and chemoresistance of A549 cells. Quantative PCR was used to detect the mRNA changes of Bcl-xL. Western blotting was used to detect the protein expression of Bcl-xL. ELISA was applied to detect the SDF-1 secretion from normal fibroblasts (NF) and CAF. Results: CAF promoted the proliferation of A549 cells, while NF had no significant effect on them. After 72 hrs incubation, the absorbance value of A549+ CAF medium group was 0.814±0.006, significantly different from the 0.753±0.006 of the A549+ NF medium group (P<0.05). The Q-PCR assay indicated that mRNA expressions of Bcl-xL in the A549 group, A549+ NF medium group and A549+ CAF medium group were 1.00±0.11, 1.10±0.09 and 3.50±0.30, respectively, showing a significant difference between the A549+ NF medium group and A549+ CAF medium group (P<0.05). The Western blot showed that protein expressions of Bcl-xL in the A549 group, A549+ NF medium group and A549+ CAF medium group were 1.00±0.08, 1.10±0.12 and 3.10±0.25, respectively, with a significant difference between the A549+ NF medium group and A549+ CAF medium group (P<0.05). The ELISA results showed that the SDF-1 concentrations in the A549+ NF medium group and A549+ CAF medium group were 3.23±0.02 and 9.53±0.10, respectively, significantly different from each other (P<0.05). The MTT assay indicated that the absorbance values of OD of A549 group, A549+ AMD3100 group, A549+ NF medium group, A549+ NF medium+ AMD3100 group, A549+ CAF medium and A549+ CA Fmedium+ AMD3100 group were 0.43±0.03, 0.25±0.02, 0.48±0.03, 0.31±0.03, 0.72±0.06 and 0.45±0.03, respectively. The data of A549+ NF medium group was significantly different from that of A549+ CAF medium group (P<0.05). Conclusions: Cancer-associated-fibroblasts enhance the drug resistance of A549 cells through SDF-1 secretion, upregulating the expression level of Bcl-xL through interaction with CXCR4. Our study not only illustrates that tumor microenvironment is able to enhance drug resistance of tumor, but also provides experimental evidence for the cancer-associated-fibroblasts as a potential therapeutic target for the treatment of lung cancer.

[Therapeutic efficacy and mechanism of action of ginsenoside Rg1 in treating acute hepatic failure in mice].

Objective: To examine the regulatory effect of ginsenoside Rg1 (G-Rg1) on endoplasmic reticulum stress and its effect on hepatocellular apoptosis in carbon tetrachloride (CCl(4))-induced acute liver failure (ALF). Methods: Forty healthy, adult male C57/BL mice were randomly divided into normal saline control (NS) group, G-Rg1 blank control (G-Rg1) group, CCl(4) model (CCl(4)) group, and G-Rg1 preventive treatment (CCl(4)+G-Rg1) group, and an ALF mouse model was established by CCl(4) induction. Blood and liver specimens were collected from all mice upon sacrifice at 12 hours post-intraperitoneal injection. Serum alanine aminotransferase (ALT), serum aspartate aminotransferase (AST) and total bilirubin (TBil) levels were determined using commercial test kits. The mRNA expression of glucose-regulated protein 78 (GRP78) and C/EBP homologous protein (CHOP) was measured using real-time PCR. The protein expression of GRP78, CHOP, caspase12, and caspase3 were measured by Western blot. Histological changes in the liver were assessed by hematoxylin-eosin staining, and the expression of GRP78 and caspase3 was detected by immunohistochemistry. Hepatocyte apoptosis was determined using terminal transferase dUTP nick end labeling. Quantitative data were analyzed using one-way ANOVA, and subsequent pairwise comparisons were performed using the LSD-t method. Results: Serum ALT, AST, and TBil levels in the CCl(4)+G-Rg1 group were significantly reduced compared with those in the CCl(4) group (ALT: 691.30 ± 108.06 U/L vs 980.66 ± 110.29 U/L, F = 365.07, P < 0.05; AST: 195.40 ± 15.41 U/L vs 319.44 ± 89.32 U/L, F = 115.64, P < 0.05; TBil: 1.09 ± 0.11 mg/dl vs 1.56 ± 0.12 mg/dl, F = 211.29, P < 0.05). The relative mRNA expression of GRP78 and CHOP was significantly lower in the CCl(4) + G-Rg1 group than in the CCl(4) group (P < 0.05). The relative protein expression of caspase3, GRP78, caspase12, and CHOP was significantly reduced to different extents in the CCl(4)+G-Rg1 group compared with those in the CCl4 group (P < 0.05). The CCl(4) + G-Rg1 group showed reduced liver tissue degeneration and necrosis compared with the CCl(4) group. Furthermore, the CCl(4)+G-Rg1 group showed significantly fewer brown granules in the liver than the CCl4 group (P < 0.05), indicating that G-Rg1 preventive treatment reduced CCl(4)-induced hepatocyte apoptosis. Conclusion: G-Rg1 prophylaxis can inhibit inflammation and reduce hepatocyte necrosis and apoptosis during CCl(4)-induced ALF. Its mechanism may involve the suppression of endoplasmic reticulum stress-related signaling molecules to alleviate hepatocyte endoplasmic reticulum stress and apoptosis. The results of this study suggest that G-Rg1 may inhibit liver inflammation and hepatocyte apoptosis through multiple targets to protect liver function.

[DNT cell inhibits the growth of pancreatic carcinoma via NKG2D and MICA pathway in vivo].

Objective: To investigate the effects of natural killer group 2 member D (NKG2D) and its ligands major histocompatibility complex class Ⅰ chain-related molecules A(MICA) on DNT cell killing pancreatic carcinoma. Methods: Antibodies adsorption was used to separate DNT cell from human peripheral blood. Human pancreatic tumor models were established via implanting BXPC-3 cells into nude mice, and then mice were randomly divided mice into 3 groups, blank group, gemcitabine group and DNT group. Mice weights and mice tumor volumes were measured every 5 days. After 40 days mice were euthanized at cervical dislocation method. Tumor weights were measured. Relative tumor volume and tumor inhibition rate were calculated. Western blot and qPCR were used to detect the expressions of NKG2D and MICA in the transplanted tumors of the three groups. Results: The blank group tumor volume and weight of blank group were significantly larger than those of gemcitabine group and DNT group (858.7±35.7 mg, 251.1±19.7 mg, 278.5±17.3 mg, P<0.001), but there were no significantly difference between DNT group and gemcitabine group. The tumor inhibition rate of gemcitabine and DNT cell were 40.4% and 35.5%. Western blot and qPCR showed that MICA mRNA and protein levels in blank group were significantly higher than those in DNT group (P=0.001, P=0.003). NKG2D mRNA and protein levels in blank group were significantly lower than those in DNT cells group (P<0.001, P=0.001). Conclusion: DNT cell can significantly inhibit the growth of pancreatic carcinoma in vivo, and the mechanism may be involved in abnormal expressions of MICA and NKG2D.

[Application of means of surfaces with nonhomogeneity in estimating the incidence of pneumoconiosis].

Objective: To investigate the value of means of surfaces with nonhomogeneity (MSN) in esti-mating the incidence of pneumoconiosis. Methods: Based on the principal component analysis, all counties (districts) of Hebei Province, China, were divided into three categories according to the degree of pneumoconiosis hazards and the MSN model was used to estimate the incidence rate of pneumoconiosis and the number of pneu-moconiosis cases using the data of the incidence of pneumoconiosis in 2010. Results: With reference to the appli-cation requirements of the MSN model, results of the principal component analysis, and expert experience, the 172 counties (districts) in Hebei Province were divided into three categories with mild, moderate, and severe pneumoconiosis hazards. There were 74, 61, and 49 counties in the above categories, respectively, and 12, 12, and 25 counties were selected from them, respectively. The estimated number of pneumoconiosis cases in Hebei Province was 2105, and the incidence rate was 261.5 per hundred thousand, with a standard error of esti-mation of 389.9 per hundred thousand. Conclusion: The MSN model provides a new thought and method for es-timating the number of pneumoconiosis cases.