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ObjectiveTo explore the therapeutic mechanism of Bushen Huoxue prescription from the perspective of bone metabolism by observing the clinical efficacy of this prescription in treating femoral head necrosis （ONFH， syndrome of liver and kidney deficiency） and its influences on bone metabolism indexes： N-terminal propeptide （PINP） and β-collagen degradation product （β-CTX）. MethodSixty-six ONFH patients with the syndrome of liver and kidney deficiency in Zhengzhou Traditional Chinese Medicine Hospital of Orthopedics from December 2021 to September 2022 were selected. The patients were randomized into an experimental group and a control group by the parallel control method， with 33 patients in each group. The experimental group received Bushen Huoxue prescription orally， while the control group received Xianlinggubao Capsules orally， with a treatment cycle of 6 months. The visual analogue scale （VAS） score， Harris score， Association Research Circulation Osseous （ARCO） staging， imaging changes， quantitative scores of TCM symptoms， and serum levels of PINP and β-CTX were determined before and after treatment. The occurrence of adverse events and reactions was recorded. ResultThe total response rate in the experimental group was 83.87% （26/31）， which was higher than that （68.75%， 22/32） in the control group （Z=-2.096， P<0.05）. After treatment， the single and total scores of TCM symptoms， VAS score， and β-CTX level decreased in the two groups （P<0.05）. Moreover， the decreases in the scores of hip pain， lower limb mobility， soreness of waist and knees， and lower limb flaccidity， total score of TCM symptoms， VAS score， and β-CTX level in the experimental were larger than those in the control group （P<0.05）. After treatment， the imaging results showed no significant improvement in the two groups. The Harris score and PINP level in both groups increased after treatment （P<0.05）， and the increases were more obvious in the experimental group than in the control group （P<0.05）. No serious adverse event or adverse reaction appeared during the observation period. ConclusionBushen Huoxue prescription can relieve pain and TCM symptoms and improve the hip joint function in treating ONFH patients with the syndrome of liver and kidney deficiency. It can inhibit the development of ONFH， increase PINP， and decrease β-CTX. No obvious side effect appears during the clinical observation period， which shows that Bushen Huoxue prescription has good safety.
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Objective:To explore the therapeutic effect of anti-interleukin (IL)-12/IL-23 p40 antibody on experimental autoimmune uveitis (EAU) and its mechanism.Methods:Sixty-six SPF female C57BL/6N mice aged 6-8 weeks were selected.EAU model was established in 24 mice through immunization with the interphotoreceptor retinoid-binding protein (IRBP) 651-670.The 24 mice were sacrificed before immunization, and on the 3rd, 12th, and 18th day after immunization, with 6 at each time point.Flow cytometry was used to detect the proportion of IL-17A + interferon-γ (IFN-γ) + CD4 + T cells in the spleen, lymph nodes and eyeballs.Another 6 mice were selected to establish EAU model, and fundus images of the mice were taken with a small animal imaging instrument and optical coherence tomography (OCT) 18 days after immunization.The 6 mice were sacrificed after OCT examination and the eyeballs were collected.Hematoxylin-eosin staining was used to observe the retinal inflammation and morphological changes in tissue structure.Flow cytometry was employed to detect the proportion of IL-17A + IFN-γ + CD4 + T cells in lymph nodes.The 6 mice were divided into IL-17A + IFN-γ + highly expressed group and IL-17A + IFN-γ + lowly expressed group according to flow cytometry results, and the retinal injury was compared between the two groups.EAU model was established in another 36 mice, which were divided into anti-IL-12/IL-23 p40 group and IgG group by random number table method, with 18 mice in each group.Anti-IL-12/IL-23 p40 or IgG was injected by tail vein at a 3-day inteval according to grouping.On the 12th and 18th day after immunization, 6 mice were selected from each group to collect lymph nodes and eyeballs, and the proportion of T cell subsets was detected by flow cytometry.Eyeballs of 6 mice in each group were extracted on the 24th day after immunization and retinal damage was observed by hematoxylin-eosin staining.The induced differentiation of CD4 + T cells in vitro was assayed by flow cytometry.The expressions of IL-17 and IFN-γ were detected by enzyme-linked immunosorbent assay (ELISA) after induced differentiation of IL-17A + IFN-γ + CD4 + T cells.The relative expression levels of Th1 transcription factor T-bet and Th17 transcription factor retinoid acid-related orphan nuclear receptor γt (ROR-γt) after induced differentiation of IL-17A + IFN-γ + CD4 + T cells were detected by real-time quantitative PCR.The use and care of animals followed the ARVO statement and this study protocol was approved by an Ethics Committee of Experimental Animals of Tianjin Medical University Eye Hospital (No.TJYY2019111019). Results:There were significant differences in the proportion of IL-17A + IFN-γ + CD4 + T cells in lymph nodes, spleen and eyeballs between wild-type mice and EAU mice at the 3rd, 12th and 18th day after immunization ( H=9.642, 16.531, 10.385; all at P<0.05). Compared with before immunization, the proportion of IL-17A + IFN-γ + CD4 + T cells was significantly increased in lymph nodes of EAU mice on the 12th day following immunization and was significantly increased in spleen and lymph nodes on day 18 after immunization (all at P<0.05). Severe retinal exudation, retinal detachment, severe inflammatory cell infiltration and extensive retinal folds were detected in IL-17A + IFN-γ + highly expressed mice.Mild retinal edema, focal inflammatory cell infiltration and mild retinal folds were found in IL-17A + IFN-γ + lowly expressed mice.The proportion of CD3 and IL-17A + IFN-γ + CD4 + T cells in the eyeballs of anti-IL-12/IL-23 p40 group was lower than that in IgG group at the 18th day after immunization, and the differences were statistically significant ( t=15.304, 8.080; both at P<0.05). On day 12 after immunization, the percentage of IL-17A + IFN-γ + CD4 + T cells in anti-IL-12/IL-23 p40 group was (0.33±0.18)%, which was significantly lower than (4.83±0.45)% in IgG group ( t=15.974, P<0.001). Compared with IgG group, the percentage of Th1, Th17, IL-17A + IFN-γ + CD4 + T cells and the expression levels of IL-17, IFN-γ, T-bet, ROR-γt in anti-IL-12/IL-23 p40 group were significantly decreased, with statistical significances (all at P<0.05). Conclusions:Anti-IL-12/IL-23 p40 has a therapeutic effect on EAU by inhibiting IL-17A + IFN-γ + CD4 + T cells.
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Objective:To investigate the effect of chimney stent for reconstruction of left subclavical artery (LSA) in thoracic endovascular aotic repair (TEVAR) for acute Stanford type B aortic dissection with insufficient anchorage zone and non-thrombotic false lumen.Methods:TEVAR with chimney stent for LSA in 39 cases of acute Stanford type B aortic dissection with insufficient anchorage zone and non-thrombotic false lumen was done from Feb 2013 to Jan 2021.Results:Covered chimney stents was used in 11 cases and bare chimney stents in 28 cases. There was no postoperative stroke, left upper limb ischemia, paraplegia, hemiplegia and death. No stent migration, reverse tear and dissection rupture were observed. One bare stent was obstructed after 18 months, and all the remaining stents were patent during follow-up. The rate of immediate type Ⅰa endoleak in covered chimney stent group and bare chimney stent group were 0(0/11) and 32.1%(9/28) respectively ( P=0.04). The distance from proximal tear to LSA in covered chimney stent group, endoleak subgroup and non-endoleak subgroup in bare chimney stent were (5.1±2.3)mm, (14.4±5.2)mm and (7.8±7.0)mm respectively ( P<0.05). False lumen thrombosis was formed in endoleak subgroup 2-8 weeks after operation, and endoleak disappeared. Conclusions:There is a correlation between immediate type Ⅰa endoleak in bare chimney stent for LSA and the distance from proximal tear to LSA, covered chimney stent can reduce the incidence of immediate type Ⅰa endoleak in TEVAR for acute Stanford type B aortic dissection with insufficient anchorage zone and non-thrombotic false lumen.
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Glioblastoma is carcinogenesis of glial cells in central nervous system and has the highest incidence among primary brain tumors. Brain metastasis, such as breast cancer and lung cancer, also leads to high mortality. The available medicines are limited due to blood-brain barrier. Abnormal activation of phosphatidylinositol 3-kinases (PI3K) signaling pathway is prevalent in glioblastoma and metastatic tumors. Here, we characterized a 2-amino-4-methylquinazoline derivative XH30 as a potent PI3K inhibitor with excellent anti-tumor activity against human glioblastoma. XH30 significantly repressed the proliferation of various brain cancer cells and decreased the phosphorylation of key proteins of PI3K signaling pathway, induced cell cycle arrest in G1 phase as well. Additionally, XH30 inhibited the migration of glioma cells and blocked the activation of PI3K pathway by interleukin-17A (IL-17A), which increased the migration of U87MG. Oral administration of XH30 significantly suppressed the tumor growth in both subcutaneous and orthotopic tumor models. XH30 also repressed tumor growth in brain metastasis models of lung cancers. Moreover, XH30 reduced IL-17A and its receptor IL-17RA in vivo. These results indicate that XH30 might be a potential therapeutic drug candidate for glioblastoma migration and brain metastasis.
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Receptor activity-modulating proteins (RAMPs) are accessory molecules that form complexes with specific G protein-coupled receptors (GPCRs) and modulate their functions. It is established that RAMP interacts with the glucagon receptor family of GPCRs but the underlying mechanism is poorly understood. In this study, we used a bioluminescence resonance energy transfer (BRET) approach to comprehensively investigate such interactions. In conjunction with cAMP accumulation, Gα q activation and β-arrestin1/2 recruitment assays, we not only verified the GPCR-RAMP pairs previously reported, but also identified new patterns of GPCR-RAMP interaction. While RAMP1 was able to modify the three signaling events elicited by both glucagon receptor (GCGR) and glucagon-like peptide-1 receptor (GLP-1R), and RAMP2 mainly affected β-arrestin1/2 recruitment by GCGR, GLP-1R and glucagon-like peptide-2 receptor, RAMP3 showed a widespread negative impact on all the family members except for growth hormone-releasing hormone receptor covering the three pathways. Our results suggest that RAMP modulates both G protein dependent and independent signal transduction among the glucagon receptor family members in a receptor-specific manner. Mapping such interactions provides new insights into the role of RAMP in ligand recognition and receptor activation.
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Objective:To explore the therapeutic effects of intravenous injection of small extracellular vesicles (sEVs) derived from human umbilical cord mesenchymal stem cells (MSCs) on experimental autoimmune uveitis (EAU) in mice.Methods:MSCs from human umbilical cord were cultured and the supernatant was collected.The sEVs were isolated by ultracentrifugation method and a NanoSight instrument was used to analyze the particle size.The expression of surface markers sEVs, CD9, CD81 and CD63 was determined via Western blot.The morphology of sEVs was observed with a transmission electron microscope.Forty-eight 7-week-old female C57BL/6 mice were seclected to establish the EAU model through immunization with interphotoreceptor retinoid-binding protein peptide 651-670 (IRBP 651-670). The mice were divided into sEVs treatment group and phosphate buffer solution (PBS) control group using a random number table, with 24 mice in each group.The mice in the sEVs treatment group were injected with 50 μg of MSCs-derived sEVs via tail vein on the 11th day after modeling.In the PBS control group, the mice were injected with the same volume of PBS.Six mice in each group were randomly selected to observe the inflammation of the retina after mydriasis with an ophthalmoscope every other day from 8th day following modeling and the inflammation scores were evaluated.Six mice were randomly selected and sacrificed on the 14th day and 6 on the 18th day following modeling in each group, and both eyeballs of the mice were enucleated.Retinal tissue sections of the 6 mice sacrificed on the 18th day were stained with hematoxylin-eosin and the pathological scores were evaluated.The infiltration of helper T 1 (Th1) cells and Th17 cells in the eyeballs of the 6 mice sacrificed on the 18th day following modeling was detected by flow cytometry.T cells were isolated from spleen and lymph nodes of the 6 mice sacrificed on the 14th day, and the proliferation of T cells under different concentrations of IRBP 651-670 (0, 1, 10 and 20 μg/ml) was detected using a 5-bromodeoxyuridine (BrdU) method.To further study the effects of MSCs-derived sEVs on Th1/Th17 cells differentiation, naive T cells of spleen from another 3 normal mice were isolated by magnetic bead negative sorting and incubated with 10 μg/ml MSCs-derived sEVs or 10 μg/ml PBS, and then were cultured under Th1/Th17 cell differentiation conditions, respectively.Flow cytometry was used to measure the differentiation of naive T cells into Th1/Th17 cells.This study protocol complied with the regulations of the care and use of laboratory animals in China and was approved by an Ethics Committee of Tianjin Medical University (No.TJYY2019103022). Results:The isolated human MSCs-derived sEVs was with an average diameter of (102.4±33.6) nm and showed a double-layer membrane vesicle structure under the transmission electron microscope.The CD9, CD63 and CD81 proteins were highly expressed in sEVs.The inflammation scores of the sEVs treatment group were significantly lower than those in the control group on day 14, 16, 18, 20 and 22 after modeling (all at P<0.05). The pathological score of mice in the sEVs treatment group was significantly lower than that of PBS control group on the 18th day following modeling ( P<0.05). The flow cytometry results showed that on day 18 after modeling, the proportions of Th1 and Th17 cells in eyeballs in the sEVs treatment group were (15.55±2.03)% and (15.67±2.15)%, respectively, which were significantly lower than (21.35±0.72)% and (20.90±1.10)% in the PBS control group ( t=6.58, 5.31; both at P<0.01). BrdU results showed that when the IRBP 651-670 concentration was 20 μg/ml, the T cell proliferation ability in the sEVs treatment group was inhibited obviously compared with the control group ( P<0.05). The proportions of naive T cells differentiated into Th1 cells and Th17 cells in the sEVs treatment group were (28.15±1.32)% and (11.60±2.23)% respectively, which were significantly lower than (31.58±1.75)% and (23.52±1.76)% of the PBS control group, and the differences were statistically significant ( t=3.93, 10.26; both at P<0.05). Conclusions:Intravenous injection of human umbilical cord MSCs-derived sEVs can reduce the inflammation in EAU mice.The mechanism may be related to inhibiting the differentiation of naive T cells to Th1 and Th17 cells, and reducing the infiltration of Th1 and Th17 cells in the eyeballs.
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Objective:To conduct theoretical analysis and experimental research on peripherally inserted venous catheters, establish theoretical models of interaction between different materials catheters and simulated skin tissues, and test different catheters at the same time to provide theoretical basis and experimental basis for the optimization design.Methods:According to the mechanical properties of the catheter at 25 ℃ and 37 ℃, a finite element model of the catheter and simulate skin tissue was established. The relationship between catheter folds and material and structure during puncture was analyzed, and the stiffness, radiographic properties, etc. were tested experimentally.Results:The performance of the catheter at different temperatures is closely related to its material. The wrinkle situation of the catheter is related to the catheter material and the inclination of the wedge surface. The elastic modulus of the polyurethane (PU) catheter is about 500 MPa and 250 MPa, respectively at room temperature (25 ℃) and body temperature (37 ℃), which meets the clinical needs of high rigidity during puncture and soft material during indwelling. When the catheter structure is the same, the PU catheter is less prone to wrinkles than the fluorinated ethylene propylene copolymer (FEP) catheter. When the catheter material is the same, the smaller the inclination of the wedge surface, the less likely the catheter to wrinkle.Conclusions:Appropriately reducing the inclination of the wedge-shaped surface of the needle of peripherally inserted venous catheters can improve the success rate of puncture. The PU catheters have good mechanical properties, they are not prone to wrinkles during puncture, and their stiffness can be reduced at body temperature, which can not only increase the success rate of puncture, but also reduce the occurrence of complications. Therefore, PU catheters have a better clinical application prospect.
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Objective:To investigate the incidence and risk factors of human cytomegalovirus (HCMV) reactivation in immunocompetent severe pneumonia patients with mechanical ventilation and their effects on clinical outcomes.Methods:A prospective observational study was conducted. Forty-eight immunocompetent patients requiring invasive mechanical ventilation due to severe pneumonia in the department of critical care medicine of the First Affiliated Hospital of Guangzhou Medical University from June 30th, 2017 to July 1st, 2018 were enrolled. Meanwhile, all cases were followed up until 90 days after inclusion and were required to quantitatively detect HCMV DNA in serum at regular weekly intervals until 28 days after transferring to intensive care unit (ICU). Patients were divided into HCMV reactivation group (≥5×10 5 copies/L) and non-reactivation group (<5×10 5 copies/L) based on HCMV DNA at any time point within 28 days. Demographic data, basic indicators, respiratory indicators, disease severity scores, laboratory indicators, complication and clinical outcomes of the two groups were collected and analyzed. Multivariate Logistic regression analysis was performed to screen independent risk factors for HCMV reactivation. Results:All 48 subjects were tested positive for HCMV immunoglobulin G (IgG), so HCMV seropositive rate was 100%. HCMV reactivation occurred in 10 patients within 28 days after admission to ICU, and the reactivation incidence of HCMV was 20.83%. There was no significant difference in gender, age, body mass index (BMI), underling disease reasons for ICU transfer (except sepsis), basic vital signs, disease severity scores, or laboratory findings including infection, immune, blood routine, liver, kidney and circulatory indicators except neutrophils count (NEU), hypersensitivity C-reactive protein(hs-CRP), hemoglobin (Hb), blood urea nitrogen (BUN), N-terminal pro-brain natriuretic peptide (NT-proBNP) between the two groups. The height (cm: 160±6 vs. 166±8), body weight (kg: 49.4±11.2 vs. 57.6±10.5), Hb (g/L: 87±18 vs. 104±24) in HCMV reactivation group were significantly lower than non-reactivation group, as well as NEU [×10 9/L:12.7 (9.9, 22.5) vs. 8.9 (6.2, 13.8)], hs-CRP [mg/L: 115.5 (85.2, 136.6) vs. 39.9 (17.5, 130.2)], BUN [mmol/L:13.7 (8.9, 21.5) vs. 7.1 (4.9, 10.5)] and NT-proBNP [ng/L: 6 751 (2 222, 25 449) vs. 1 469 (419, 4 571)] within 24 hours of admission to ICU. The prevalence of sepsis [60.0% (6/10) vs. 15.8% (6/38)], blood transfusion [100.0% (10/10) vs. 60.5% (23/38)], hospitalization expense [ten thousand yuan: 35.7 (25.3, 67.1) vs. 15.2 (10.4, 22.0)], 90-day all-cause mortality [70.0% (7/10) vs. 21.1% (8/38)], length of ICU stay [days: 26 (16, 66) vs. 14 (9, 19)], the duration of mechanical ventilation [days: 26 (19, 66) vs. 13 (8, 18)] in HCMV reactivation group were significantly higher than non-reactivation group, and there were significant statistical differences between the two groups (all P < 0.05). Logistic regression analysis showed that sepsis was an independent risk factor for HCMV reactivation in immunocompetent mechanical ventilation severe pneumonia patients with mechanical ventilation [odds ratio ( OR) = 9.35, 95% confidence interval (95% CI) was 1.72-50.86, P = 0.010]. Conclusions:HCMV infection is very common in immunocompetent severe pneumonia patients on mechanical ventilation and incidence of HCMV reactivation is high. Moreover, HCMV reactivation could adversely affect clinical prognoses, and sepsis may be a risk factor for HCMV reactivation.
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Objective To observe the image characteristics ofmultispectral scanning laser imaging (MSLI) and OCT in patients with pregnancy induced hypertension syndrome (PIHS).Methods A total of 112 patients (224 eyes) of PIHS patients diagnosed in Obstetrics Department of Tianjin First Central Hospital from May 2016 to May 2017 were included in this study.The average age of the patients was 27.00±2.14 years.The average course of the disease was 15.00 ±8.27 days.There were 174 eyes in 87 patients of blurred vision,dazzling and visual fatigue consciously.All patients performed BCVA,direct ophthalmoscope,B ultrasound,confocal scanning laser Ophthalmoscope (cSLO) and spectral-domain OCT (SD-OCT).SD-OCT was performed with Spectralis HRA+OCT from Heidelberg Company in Germany to acquire tomographic images.Using Herdelberg's colorful program (MultiColor) based on cSLO and operating in accordance with standard methods,one scan simultaneously obtained blue light reflection based on 488 nm,green light reflection based on 515 nm,and infrared reflection based on 820 nm,synthesis to MSLI.Fundus abnormalities were classified into arterial spasm (stage Ⅰ),arteriosclerosis (stage Ⅱ),and retinopathy (stage Ⅲ).OCT examination was divided into normal and abnormal cases according to the abnormality of retinal morphology and thickness.Results Of the 224 eyes,68 eyes (30.36％) showed normal fundus examination and 156 eyes (69.64％) showed abnormal fundus performance.Among them,28 eyes were stage Ⅰ (17.95％);40 eyes were stage Ⅱ (25.64％);88 eyes were stage Ⅲ (56.41％).Thirty-six eyes (16.07％) showed normal fundus and 188 eyes (83.93％) showed abnormal performance with OCT.Of the 188 eyes with abnormal fundus performance,86 eyes (45.74％) had retinal neuroepithelial serous detachment;56 eyes (29.79％) had RPE detachment;optic disc edema,bulge,and local reflexes in the retinal nerve fiber layer were enhanced and/or the thickness increased in 46 eyes (24.47％).In MSLI,48 eyes (21.43％) showed normal fundus;176 eyes (78.57％) showed abnormal performance.Retinal edema was showed in green on MSLI,serous retinal neuroepithelial layer detachment,RPE layer detachment,retinal nerve fiber layer thickening,accompanied by changes in local retinal structure.The higher the degree of bulge,the darker the color.Consistent with the range of retinal edema revealed by SD-OCT.Conclusions MSLI and SD-OCT images show highly consistent lesions in PIHS patients.MSLI can more clearly show superficial and deep retinal lesions.
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Objective:To investigate the association of the difference between 2-hour postprandial blood glucose and fasting blood glucose with non-alcoholic fatty liver disease(NAFLD)in elderly patients with type 2 diabetes(T2DM).Methods:A total of 953 patients aged≥65 years with T2DM hospitalized in the Department of Endocrinology and Metabolism, Shanghai Jiaotong University Affiliated Sixth People's Hospital from January 2007 to June 2009, with complete clinical data, were selected in this retrospective study.The absolute value of the difference between 2-hour postprandial blood glucose and fasting blood glucose was recorded as the blood glucose difference.According to the tertiles of the blood glucose difference, patients were divided into three groups including the first tertile(n=317, blood glucose difference<4.12 mmol/L), the second tertile(n=320, blood glucose difference between 4.12-7.69 mmol/L), and the third tertile(n=316, blood glucose difference≥7.69 mmol/L). Detailed clinical data of the patients were collected, and clinical characteristics and the prevalence of NAFLD were compared among the three groups.The correlation between the blood glucose difference and NAFLD was analyzed.Results:From the first tertile to the third tertile, there appeared to be increased proportions of patients using metformin( χ2=9.581, P=0.008), higher waist-to-hip ratios( F=3.663, P=0.026), increased 24 h uric acid excretion( χ2=6.241, P=0.044), increased alanine aminotransferase levels( χ2=22.361, P<0.001), increased γ-glutamyl transpeptidase levels( χ2=17.681, P<0.001)and increased 2h postprandial blood glucose levels( χ2=579.315, P<0.001), with significant statistical differences in the waist circumference( F=4.723, P=0.009), body mass index( F=5.811, P=0.003), fasting C-peptide levels( χ2=9.442, P=0.009), 2h postprandial C-peptide levels( χ2=17.599, P<0.001)and glycosylated hemoglobin A1c( F=30.836, P<0.001)between the three groups and the prevalence of NAFLD also grew steadily(24.0%, n=76 vs. 33.1%, n=107 vs. 36.7%, n=116, χ2=12.712, P<0.01). After adjusting for other variables, multivariate logistic regression analysis showed that the blood glucose difference was correlated with NAFLD in elderly patients with T2DM( OR=1.396, Wald χ2=0.002, P<0.01). Conclusions:Elderly T2DM patients with greater blood glucose differences have more severe metabolic disorders and a higher prevalence of NAFLD.An increase in blood glucose differences is an independent risk factor for NAFLD in elderly T2DM patients.
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Objective@#To explore the feasibility of bronchoscopic brushing liquid-based slide cytology combined with automatic immunocytochemistry (ICC) for pathological typing of lung cancer.@*Methods@#A liquid-based thin-prep was prepared from 171 bronchoscopic brushing specimens of patients with pulmonary lesions. ICC was detected by automatic immunohistochemistry instrument while cytomorphological diagnosis was made. The results were compared with those of histopathological diagnosis.@*Results@#Among 171 patients, 130 (76.0%) could be classified by cell morphology alone, including 31 squamous cell carcinomas, 44 adenocarcinomas and 55 small cell carcinomas; 162 (94.7%) could be classified by cell morphology combined with ICC, including 38 squamous cell carcinomas, 61 adenocarcinomas and 63 small cell carcinomas (P<0.001). According to the gold standard of histopathological diagnosis, the coincidence rate of cytomorphology combined with ICC was higher than that of cell morphology alone. The coincidence rate of squamous cell carcinoma was increased from 85.2% to 97.1% (P=0.093), adenocarcinoma from 92.5% to 98.0% (P<0.001), and small cell carcinoma from 96.1% to 98.3% (P=0.465).@*Conclusion@#The combination of liquid-based thin-prep cytology and automatic immunohistochemistry can effectively improve the accuracy of pathological typing of brushing specimens under fiberoptic bronchoscopy, and provide more objective diagnostic results for clinical treatment.
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Objective@#To evaluate the utility of the BRAFV600E mutation detection in different cytoloy categories and to determine if the VE1 antibody can serve as a screening tool for the detection of BRAFV600E mutation in thyroid fine needle aspiration (FNA) specimens.@*Methods@#A total of 273 FNA residual specimens were collected. BRAFV600E testing was performed on these liquid-based specimens. And also 78 specimens with enough residual cells were stained with VE1 antibody. Comparisons of molecular and immunocytochemistry results with clinicopathological outcomes were performed. SPSS 17.0 software was used to analyze the data.@*Results@#There were 70 indeterminated diagnoses in 273 cases with FNAs. Fifty-eight cases were proven to be papillary thyroid carcinoma (PTC) by histology, including 9 cases of atypia of undetermined significance/follicular lesion of undetermined significance (AUS/FLUS), 3 cases of follicular neoplasm/suspicious for follicular neoplasm (FN/SFN), and 46 cases of suspicious for malignancy (SM). BRAFV600E analysis detected PTC in 3 of 9 cases with AUS/FLUS, and in 31 (67.4%) of 46 cases with SM. The sensitivity of immunostaining with VE1 antibody was 62.8%(27/43) and the specificity was 91.4% (32/35). VE1 expression showed moderately concordance with the molecular mutation (κ=0.524, P<0.001).@*Conclusions@#Additional BRAFV600E detecting can improve the diagnostic efficacy for PTC in AUS/FLUS and SM. VE1 expression may be an alternative method for BRAFV600E detecting when molecular detecting is unavailable.
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Objective@#To observe the expression of retinal proteins in experimental autoimmune uveitis (EAU) mice and to explore the possible molecular mechanism of autoimmune uveitis.@*Methods@#Twelve female C57BL/6J mice were randomly divided into model group and normal control group, 6 mice in each group.In the model group, the EAU model was established by subcutaneous injection of human interphotoreceptor retinoid-binding protein (IRBP) 651-670.The fundal change of EAV mice was assessed by direct ophthalmoscope, OCT and histopathological staining.At 18 days after immunization, the retinas of the two groups were taken for retinal protein extraction, protein restriction enzyme digestion, mass spectrometry detection, data analysis, and bioinformatics analysis.This study was approved by the experimental animal Ethics Committee of Tianjin Medical University Eye Hospital (TJYY2018070113). The feeding and use of experimental animals follow the ARVO statement.@*Results@#The EAU mouse model was successfully established.At 10 days after immunitation, the retina of EAV mouse was damaged.At 18 days after immunization, retinal edema and infiltration of inflammatory cells into vitreous were observed.Proteomic results showed that a total of 4 458 proteins were identified in this study, of which 522 were differentially-expressed proteins (fold change>1.5, P<0.05). Among these differentially-expressed proteins, 147 proteins including Stat1 and Stat3 were up-regulated and 375 proteins including Gucy2f and Rho were down-regulated.These differentially-expressed proteins were closely related to platelet activation, integrin signaling, T cell activation, the phototransduction cascade, Toll-like receptor and so on.@*Conclusions@#EAU is related to the abnormal expression of Stat1, Stat3 and other proteins, as well as the abnormal regulation of platelet activation and integrin signal pathway.
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OBJECTIVE： To explore the effects of different primary processing methods on quality change of Polygonatum sibiricum， and to optimize primary processing method of P. sibiricum with different grades and shapes. METHODS： After processed with different drying methods， boiling method and steaming method， sulfuric acid-phenol method was used to determine the contents of indicator component P. sibiricum polysaccharide in samples. The contents of ethanol-soluble extract， water-soluble extract and total ash were investigated. The optimal primary processing methods were screened. RESULTS： After processed with different methods， the contents of polysaccharide （≥7％）， ethanol-soluble extract （≥45％） and total ash （≤4％） of samples at all levels conformed to the requirements of the 2015 edition of Chinese Pharmacopoeia （part Ⅰ）； from aspect of drying method， microwave drying method was better than oven drying method；for P. sibiricum with different morphologies （individual and lamellar）， the quality of lamellar samples of P. sibiricum was better than that of individual P. sibiricum. As primary processing method， steaming was better than boiling. The contents of P. sibiricum polysaccharide were in relatively high level， when the steaming and boiling time of gradeless and uniformly-priced goods of P. sibiricum were 20 and 15 minutes respectively. CONCLUSIONS： By discussing the different primary processing methods， high-efficient primary processing method for the individual and lamellar samples of P. sibiricum as well as gradeless and uniformly-priced goods of P. sibiricum are preliminarily obtained， which can provide a scientific basis for the effective utilization of P. sibiricum resources.
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Objective@#To explore the feasibility of conventional smears and liquid-based cytologic slides of lymphatic metastasis specimens of lung adenocarcinoma acquired by fine needle aspiration cytology (FNAC) to detect the expression of anaplastic lymphoma kinase (ALK/D5F3) by immunocytochemistry (ICC) analysis.@*Methods@#The lymphatic metastasis specimens of 147 lung adenocarcinoma, including 100 liquid-based cytologic slides and 47 conventional smears, were collected in this study. ALK fusion protein was detected by Roche Ventana ICC technology, which was compared with the ALK fusion gene assessed by fluorescence in situ hybridization (FISH) or reverse transcriptase-polymerase chain reaction (RT-PCR).@*Results@#The positive rate of ALK (D5F3) fusion protein in advanced lung adenocarcinoma acquired by FNAC was 11.6% (17/147), and 10.6% (5/47) and 12.0% (12/100) were reached in conventional smears and liquid-based cytologic slides, respectively.Among 147 cases, 57 cases including 17 positive cases and 40 negative cases were verified by RT-PCR and FISH. The whole coincidence rate reached 96.5% (55/57). The sensitivity and specificity of ALK (D5F3) fusion protein detected in lung adenocarcinoma acquired by FNAC were 94.1% (16/17) and 97.5% (39/40), respectively. The sensitivity and specificity were both 100% (5/5 of sensitivity and 10/10 of specificity) in conventional smears, while 91.7% (11/12) and 96.7% (29/30) in liquid-based cytologic slides.@*Conclusion@#Conventional smears and liquid-based cytologic slides of FNAC samples can be used to perform ICC analysis of ALK (D5F3) expression in advanced lung adenocarcinoma, especially for patients who have no opportunity for surgery or whose resected samples are difficult to form cell block.
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Objective@#To analyze the composition and richness of intestinal microflora in children with non-alcoholic fatty liver disease (NAFLD) and the role of which in pathogenesis of NAFLD.@*Methods@#This was a prospective case-control study. From November 2015 to June 2017, 19 children diagnosed with NAFLD according to the 2010 edition of diagnostic criteria were enrolled voluntarily in the Second and First Affiliated Hospitals of Zhejiang Chinese Medicine University. Twenty-two healthy children were enrolled in the control group. Among the patients, 10 were males and 9 were females, at the mean age of (11.0±1.0) years; 10 males and 12 females in the control group, at the mean age of (9.0±1.2) years. The body mass index (BMI) and waist circumference were recorded, and the fasting blood glucose, total cholesterol, triglyceride, high-density lipoprotein and low-density lipoprotein were detected. Feces were collected and the fecal microorganisms were extracted with magnetic beads methods; the composition and the richness of intestinal microflora in the two groups were detected with 16S rDNA high throughput sequencing technology. The KO differential gene expression and KEGG signal pathway enrichment were analyzed with PICRUST software. The intestinal flora characteristics between the two groups were compared with t test or Mann-Whitney U test and Willcoxon W test.@*Results@#The BMI, waist circumference and triglyceride were higher in NAFLD group than those in the control group (BMI (25.1±2.7) vs. (18.2±1.5)kg/m2, t=9.912, P=0.000; waist circumference (88.6±6.6) vs. (71.5±6.3) cm, t=8.520, P=0.000; triglyceride (0.9±0.4) vs.(0.7±0.3)mmol/L, t=2.060, P=0.046). The abundance and diversity index of intestinal microflora were lower in the NAFLD group (Shannon index 3.99 (3.13, 4.54) vs. 4.63 (4.21, 4.81), Z=-2.065, P=0.039; Simpson index 0.85 (0.70, 0.89) vs. 0.90 (0.88, 0.93), Z=-2.431, P=0.015; ACE index 235.76 (205.26, 361.94) vs. 326.96 (275.34, 368.65), Z=-2.092, P=0.036). At the level of phylum, the proportion of Actinomycetes was lower and the proportion of Thermus was higher in NAFLD group (Actinobacteri 29.807 (14.723, 62.080) ×10-3 vs. 63.212 (46.133, 172.071) ×10-3, Z=-2.667, P=0.008; Thermus 0.033 (0.000, 0.226) ×10-3 vs. 0.000 (0.000, 0.031) ×10-3, Z=-2.729, P=0.006) . At the level of genus, the proportion of Bacteroides and Bifidobacterium in the NAFLD group were significantly lower (Bacteroides 78.757 (11.430, 151.621) ×10-3 vs. 356.821 (161.049, 403.037) ×10-3, Z=-2.771, P=0.006; Bifidobacterium 19.680 (6.181, 53.944) ×10-3 vs. 54.721 (31.911, 146.410) ×10-3, Z=-2.458, P=0.014); the proportion of Prevotella in NAFLD group was significantly higher (3.089 (0.165, 63.502) ×10-3 vs. 0.432 (0.029, 2.257) ×10-3, Z=-2.112, P=0.035). Based on the KEGG database, 78 differentially expressed genes and 26 differential metabolic pathways were found, among which the function genes of K01470, K01961 and K07258 were concentrated in the pathways of arginine and proline metabolism, fatty acid synthesis, and polysaccharides biosynthesis and metabolism. Besides, these three function genes were related to Bacteroides, Prevotella, Bifidobacterium and Ruminococcus.@*Conclusion@#NAFLD children have intestinal flora disturbances in both diversity and abundance, which may alter lipid metabolic pathways through differential gene expressions, contributing to the pathogenesis of NAFLD.
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Objective To establish the allele-specific real-time polymerase chain reaction (ASPCR) for detection of neonatal hyperbilirubinemia related gene SLCO1B1 A388G polymorphism and apply this assay to identify the clinical samples.Methods According to SLCO1B1 A388G polymorphism loci,specific primers were designed and the assay was established.Wide type plasmid and mutant plasmid were constructed.Fifty clinical samples were selected,including 30 samples of neonatal hyperbilirubinemia that had been diagnosed with SLCO1B1 A388G mutant and 20 samples of healthy newborns without SLCO1B1 A388G mutant were selected as the controls.Wide type plasmid,mutant plasmid and clinical samples were tested by specific and non-specific primers.A388G polymorphism was determined by difference in Ct (cycle threshold) between specific and non-specific primers.Then,the accuracy,sensitivity and specificity of assay were evaluated.Results The difference in Ct (cycle threshold) between specific and non-specific primers that amplified equivalent wide type template was 13.97 ±0.75.The assay could correctly distinguish the wide type and mutant plasmid.Probit regression analysis showed the sensitivity of the assay could reach to 5.28 copies/μL.For clinical samples,the Ct values of the samples with A388G mutation was less than 37.75 and showed positive results,while the samples without A388G mutation did not show any amplification nor Ct values were larger than 37.75,which showed negative results.Conclusions ASPCR is a fast,simple and effective method for SLCO1B1 A388G polymorphism detection of the clinical simples.It can be used for large sample screening for neonatal hyperbilirubinemia gene loci.
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The development of the healthcare industry puts forward higher and higher standards on the medical education, so an effective education management mode is needed urgently for medical schools to promote the cultivation of undergraduate medical students. Tutorial system has a positive effect in building the innovative thinking, critical thinking and clinical thinking of medical students. It also helps students' innovation ability training. This paper first introduces the overview of the implementation of tutorial system in medical colleges, and then expounds the specific implementation measures of the undergraduate tutorial system in the Third Military Medical University. Investigation data shows that, accompanied with these mea-sures, the numbers of article publishes and the achievements in various competitions have been raised whereas the psychological counseling needs decreased, indicating that the tutorial system has a strong posi-tive influence on the development of medical students.
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Objective To investigate the clinical effect of Tiotropium Bromide Powder for Inhalation combined with mental intervention in the treatment of chronic obstructive pulmonary disease in stable period. Methods 80 patients with stable chronic obstructive pulmonary disease treated in our hospital from January 2015 to January 2017were divided into two groups according to the different treatment methods.The control group was treated with conventional methods, the observation group was treated with routine + Tiotropium Bromide Powder for Inhalation + psychological intervention therapy.The clinical curative effect and the improvement of clinical symptoms were recorded and analyzed. Results The clinicalcurative effect of the observation group was better than that of the control group, the improvement of clinical symptoms was better than that of the control group,the difference was statistically significant (P<0.05). Conclusion The effect of chronic obstructive pulmonary disease patients choose conventional treatment+Tiotropium Bromide Powder for Inhalation + psychological intervention mode is significant, can improve clinical symptoms, improve the therapeutic effect and prognosis in the near future and far, worthy of clinical widely used In the treatment of patients with chronic obstructive pulmonary disease in stable phase.
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AIM To observe the effects of Guhoukang Granules (Eucommiae Cortex,Dipsaci Radix,Corni Fructus,etc.) on bone metabolism indexes of the geriatric femoral intertrochanteric fracture.METHODS A total of eighty patients meeting the inclusion criteria,cured with fixed operation of proximal femoral anti-rotation nail and given basic care,were divided randomly and equally into two groups:experiment group (Guhoukang Granules ) and control group (calcium carbonate D3 tablets).Three months made a course.Osteocalcin (OC),calcitonin (CT),bone-specific alkaline phosphatase (BALP),total procollagen type 1 aminoterminal propeptide (tPINP),β isomer of the C-terminal telopeptide of type Ⅰ collagen (β-CTX),and intact parathyroid hormone (iPTH),were examined before and after three months,respectively.RESULTS There were no significant statistical difference in the levels of OC,CT,BALP,tPINP,β-CTX and iPTH before operation in both groups.The levels of OC,CT,BALP,tPINP in the experiment group obviously increased and the levels of β-CTX,iPTH markedly decreased after the operation,and were better than those in the experiment group pre-operation and in the control group after the operation.The control group had no improvement in bone metabolism indexes pre-and post-operation.CONCLUSION Guhoukang Granules can improve bone metabolism of the geriatric femoral intertrochanteric fracture and facilitate the recovery of the fracture.