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Effective supplementation of probiotics can be beneficial to intestinal health, but in situ analysis of probiotics activity has rarely been reported. In this study, by coupling fluorescein 5-isothiocyanate (FITC) and 5(6)-carboxytetramethylrhodamine N-succinimidyl ester (5(6)-TAMRA-SE) with D-lysine, two fluorescent D-amino acids (FDAAs) probes were obtained: green probe (fluorescein-D-lysine, FDL) and red probe (TAMRA-D-lysine, TDL). Then, we tried to label the three kinds of probiotics, Lactobacillus acidophilus (LA), Lactobacillus casei (LC) and Veillonella atypica (VA) in vitro. FDAAs was applied to the labeling of intestinal flora in mice, and a method was established to investigate the oral survival rate of three commonly used probiotics. All animal experiments were approved by the Animal Ethics Committee of Guangzhou University of Chinese Medicine. The results show that the two synthetic FDAAs can be non-toxic and 100% for the in vitro labeling of the three probiotics. Known by FDAAs two-step labeling of oral probiotics, the high survival rate of LA was 92.30% ± 1.67%. The survival rates of VA and LC are similar, 84.13% ± 4.06% and 82.27% ± 2.43%, respectively. This study can quickly compare the changes of colonization survival rate of different probiotics in vivo, provide theoretical support for the in situ colonization activity of probiotics in the intestine, and guide the rational drug use of clinical probiotics.
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Objective@#To analyze myopia related factors among students in Shanghai, to explore associated risky behaviors associated with mild, moderate, high myopia,so as to provide the evidence for the implementation of home and school combination intervention measures.@*Methods@#Six primary and secondary schools (2 primary schools, 2 middle schools and 2 high schools) were selected from each district of Shanghai. The primary schools began to investigate from the fourth grade. All the students in the selected classes participated in the vision test and questionnaire survey. Chi square test was used for categorical data analysis. The relationship between myopia and related behaviors was analyzed by multivariate Logistic analysis.@*Results@#The prevalence of visual-related risky behaviors such as short outdoor time during the day, lack of sleep, long after-school reading and writing time, poor reading and writing posture, and longtime-using mobile electronic screen was higher in girls than in boys (P<0.05), boys were more likely than girls to use computer for long time and read books/electronic screen in sunlight (P<0.05). The group with 3 hours or more than of reading and writing compared with the less than 2 hours, the OR value of mild myopia model was 1.31(1.20-1.44), moderate myopia model was 1.78(1.62-1.96), severe myopia model was 2.37(2.07-2.71). In the model of moderate and high myopia, reading and writing posture, frequency of eye relax, outdoor activity time and watching TV time were also included.@*Conclusion@#The prevalence of myopia related behaviors among primary and secondary school students is high, and there are significant gender differences among different behaviors. Students reading and writing time should be strictly controlled after school. Intervention strategies and measures should be carried out according to the characteristics of different ages and genders.
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Objective To explore the relationship among post-traumatic stress disorder(PTSD),social support and quality of life in patients with spinal cord injury(SCI)after a long recovery in China and investigate the factors influencing the quality of life. Methods In this cross-sectional study,206 SCI patients who were hospitalized in 9 hospitals were enrolled.Data collection was performed using general information,the Post-Traumatic Stress Disorder Checklist-Civilian version,the social support scales,and the World Health Organization Quality of Life-Abbreviated version.The data were statistically analyzed using
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Humans , China/epidemiology , Cross-Sectional Studies , Quality of Life , Social Support , Spinal Cord Injuries/psychology , Stress Disorders, Post-Traumatic/epidemiologyABSTRACT
The present study is to establish the fingerprints for the quality evaluation of Ilicis Pubescentis Radix by HPLC-UV. The chromatographic conditions were defined as Phenomenex Luna C₁₈(4.6 mm × 250 mm, 5 μm). Mobile phase was acetonitrile-0.05% phosphoric acid in gradient elution, and the flow rate was 0.8 mL·min⁻¹.Column temperature was 30 °C and the injection volume was 10 μL．The detection wavelength was 210 nm. According to the similarity evaluation, the chemometric method was used to assess the quality of Ilicis Pubescentis Radix. The fingerprints of 16 batches of Ilicis Pubescentis Radix were established. There were 29 common peaks in the fingerprints and 12 common peaks were identified by reference substances. Fingerprints similarity of samples were greater than 0.92. The samples were classified into three groups by hierarchical cluster analysis combined with principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA), and seven components were the main markers that cause differences in the different batches of samples. By comparing the on-line UV spectra of chromatographic peaks, the chromatographic fingerprint was divided into three regions: region A showed seventeen main peaks (mainly lignans and phenolic acids); region B showed eight main peaks, which were proved as saponins; region C showed four main peaks, which were proved as other components. The established HPLC-UV fingerprint is highly specific, and can be used to evaluate the quality consistency of different batches of Ilicis Pubescentis Radix.
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Objective To explore the association of nocturnal serum cortisol levels with diabetic microvascular complications in overweight or obese patients with type 2 diabetes mellitus. Methods Serum cortisol levels of 316 overweight or obese type 2 diabetic patients were tested at midnight by the method of chemiluminescence. Diabetic microvascular complications were compared among various groups according to nocturnal serum cortisol levels. All the patients with nocturnal serum cortisol level > 50 nmol/L were asked to undergo overnight low-dose dexamethasone suppression test to rule out the possibility of subclincal Cushing's syndrome. The incidences of diabetic nephropathy ( DN ) , diabetic retinopathy ( DR ) , and diabetic peripheral neuropathy ( DPN ) were examined in all the patients. Results (1)The incidence of DN was gradually increased from 13.3％to 27.7％and 44.2％in patients with low, medium, and high cortisol level groups, showing a statistical difference among 3 groups ( P<0.05) . The incidences of DR in medium and high cortisol level groups were higher than that in low cortisol level group (40.6％and 47.7％vs 22.7％, both P<0.01). The incidence of DPN in high cortisol level group was higher as compared with low cortisol level group (60.5％ vs 38.7％, P<0.01). (2) Nocturnal serum cortisol level in patients with diabetic microvascular complications was higher than that in patients without complications [ (136.87 ± 105.78 vs 97.55 ± 93.48) nmol/L, P<0.01]. Nocturnal serum cortisol level in patients with multiple diabetic microvascular complications was higher than that in patients with single diabetic microvascular complication [ (151.66±114.54vs117.69±90.26)nmol/L,P<0.05].(3)Singlefactorlogisticregressionanalysisshowedthat higher nocturnal serum cortisol level was a risk factor for diabetic microvascular complications in addition to female, age, longer diabetic duration, higher fasting plasma glucose ( FPG ) . Multivariate logistic regression analysis showed that higher nocturnal serum cortisol level was still a risk factor for diabetic microvascular complications after adjusted by diabetic duration, FPG, HbA1C, and the use of insulin (P=0.013). Conclusion Nocturnal serum cortisol level seems to be a risk factor for diabetic microvascular complications in overweight or obese patients with type 2 diabetes mellitus.
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Objective To observe the relationship of IL-17 and arterial blood gas, fibrin fragment D (D-D) in patients with both pulmonary embolism (PE) and obstructive sleep apnea-hypopnea syndrome (OSAHS). Methods We collected 20 patients with both PE and OSAHS and 43 patients with only PE who hospitalized in the first ward of pneumology department of the Second Affiliated Hospital of Kunming Medical University, then measured D-D, IL-17 in venous blood and arterial blood gas and recorded them to analysis.Re s ults PO2 in patients with both PE and OSAHS was significantly lower than that in patients with only PE, while D-D and IL-17 of the latter were markedly lesser than the former. Conclus ions The expression of IL-17 between the 2 groups of patients has statistically significant difference. What's more, the expression of IL-17 is positively associated with hypercoagulability and Body Mass Index (BMI), and the result shows a negative relation between arterial blood oxygen partial pressure and IL-17, suggesting that IL-17 may be relate to the common progress of PE and OSASH. Thus, IL-17 can be used in the detection of patients with both PE and OSASH
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Objective:The present study aim to explore the difference and characteristics of disgust in obsessive-compulsive disorder(OCD) with/without contamination washing symptoms,adding to the growing literature on the heterogeneity and clinical treatment of OCD.Methods:Totally 66 patients with OCD meeting the criteria of International Statistical Classification of Diseases and Related Health Problems,Tenth Revision (ICD-10) and 51 healthy controls matched with gender,age and level of education were recruited.All patients were divided into two subgroups namely washing symptoms group(n =26) and other symptoms group(n =40) based on the contamination washing symptoms.Participants respectively completed the lexical decision task.The results of the tasks were indicators reflecting the disgust feelings,including the accuracy,reaction time to core disgust words,moral disgust words,neutral words,and the rating intensity of disgust provoked by all of the words.Results:The reaction time for core disgust words[(723 ± 89)ms,(746 ± 95) ms vs.(676 ± 96) ms] and moral disgust words[(772 ± 98)ms,(796± 92)ms vs.(723 ± 94)ms] were longer in both group of patients with OCD than in healthy controls.The patients also rated higher degree of disgust for core disgust words[(6.7 ± 1.5),(6.9 ± 1.6)vs.(5.8 ± 1.7)]and moral disgust words [(6.8 ± 1.7),(7.2 ± 1.3)vs.(6.3 ± 1.5)] than healthy controls (Ps ＜0.05).But there were no difference existed between patients with and without contamination washing symptoms on the results of lexical decision task(Ps ＜0.05).Conclusion:It shows that patients with OCD tend to experience intense disgust feelings,and there is no difference between contamination washing symptoms and other symptoms on disgust.These findings suggest that intense disgust feelings may play a role on the etiology and maintenance of OCD,and reducing disgust could be a potential approach for OCD treatment.
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Face to various disaster and dilemma, the tasks of national rescue team were fast and effective rescue, and reducing the harm of life and damage of property, and decreasing each damage to the lowest point. The ability of national rescue team could reflect the strength and developed degree of a national, it further more manifested the levels of medical rescue team. Face to major disasters, it was vital to provide humanitarian assistance for the affected people in disaster areas at the first time. Many developed countries have established national rescue teams and equipped various kinds of rescue equipment, and the disaster relief of medical team was regarded as the main force in emergency health rescue for disaster. refer to the constitution and equipment of national rescue team of developed countries has an important guiding role for perfection of emergency medical rescue in China.
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Objective To identify the activity of HCV IRES translation differences and identify the relationship between HCV IRES translation activity and ROS in different concentrations of ferric ammonium citrate ( FAC) in-duction.Methods 1 ) Expression plasmid pCI-Rluc-HCV IRES-Fluc was confirmed by endonuclease digestion as well as luciferase transient expression in Huh-7 cell;2) Controlled by dual-luciferase reporter assay, the differ-ent translation activity of HCV internal ribosomal entry site ( IRES ) was examined in a concentration of 50 μmol/L and 300μmol/L of FAC induction;ROS fluorescent staining method was used to detect the activity of ROS in Huh-7 cells, Western blot method was used to detect the protein expression changes of Nrf2 in Huh-7 cells;3) On the basis of the above experiments, 100 μmol/L DPI was added in 300 μmol/L FAC experimental group, to analyse the changes of HCV replication and ROS production after joining DPI.Results The generation of ROS and the activity of luciferase in the model group were significantly higher than that in the control group ( P<0.05 ) .FAC can enhance the expression of HCV IRES and increase the production of ROS , then causing Nrf2 expression in Huh-7 cell.However,after adding ROS inhibitor DPI, the above functions in Huh-7 cell were weakened.Conclusions The increase of HCV IRES expression induced by FAC is related to excessive ROS pro-duction induced by FAC in Huh-7 cells.
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Objective To contrast clinical effect of normal frequency mechanical ventilation vs .nasal continuous positive air‐way pressure ventilation in the treatment of neonatal respiratory failure .Methods Sixty children diagnosed as neonatal respiratory failure were enrolled in our hospital from January 2013 to October 2014 ,and the children were randomly divided into observation group (n=31) and control group (n=29) .Observation group were given nasal continuous positive airway pressure ,children with the control group received normal frequency mechanical ventilation .The X‐ray of the two groups before treatment and 2 d after treatment were observed and scored ,the 3 d survival rate ,blood gas analysis index ,ventilator associated pneumonia ,time of opera‐tion ,and incidence of pulmonary hemorrhage were observed in the two groups .Results The X‐ray results showed that :after treat‐ment ,the two groups of children with X‐ray film scores were gradually decreased ,compared with before treatment ,the difference was statistically significant (P0 .05) . Time ,incidence of pulmonary hemorrhage and incidence of ventilator associated pneumonia in observation group were less than those in control group ,the difference was not statistically significant (P>0 .05) .Conclusion The normal frequency of mechanical ventilation and nasal continuous positive airway pressure can be effective reducing the relevance of respiratory pneumonia in the treatment of neonatal respiratory failure ,short‐term effect of nasal continuous positive airway pressure is slightly better than normal frequency mechanical ventilation .
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Objective To investigate the role of autophagy in radiation-induced death process of human esophageal squamous carcinoma Eca-109 cells.Methods Esophageal carcinoma cell line Eca-109 was divided into 6 groups of control,5 mmol/L 3-Methyladenine treatment,10 mmol/L treatment,6 Gy irradiation,irradiation + 5 mmol/L drug,and irradiation + 10 mmol/L drug.Some cells were transferred with GFP-LC3 plasmid and the changes of autophagosome were obserred.After each treatment,the expression of autophagy marker LC3B was measured by Western Blot,cell viability was detected by MTT,morphological characteristics of apoptosis cells were stained with a fluorescein of Hoechst 33342 and the percentage of apoptotic cells and cell cycle distribution were measured by flow cytometry.Clonogenic survival were used to evaluate the cell radiosensitivity.Results Autophagy level was increased after radiation,and the LC3B Ⅱ expression and LC3B Ⅱ/LC3B Ⅰ ratio were significantly decreased by autophagy inhibitor 3-Methyladenine (F =25.64,P ＜ 0.05).The number of autophagosome fluorescent foci were significantly increased in the GFP-LC3 transfected cells after radiation,but reduced by 3-Methyladenine (F =127.36,P ＜ 0.05).Compared with radiation alone group,autophagy inhibition combined with radiation significantly decreased cell viability (F =129.54,P ＜ 0.05) and colony formation,increased apoptosis and the percentage of G2/M-phase cells.Conclusions 3-Methyladenine enhances the radiosensitivity of esophageal squamous carcinoma Eca-109 cells,suggesting that inhibition of autophagy could be used as an adjuvant treatment of radiotherapy in esophageal squamous carcinoma.
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Objective To investigate the effect of noninvasive ventilator therapy on serum C-reactive protein (CRP), endothelin-1 (ET-1) and tumor necrosis factor-α (TNF-α) levels in patients with obstructive sleep apnea hypopnea syndrome (OSAHS) and its clinical significance. Methods One hundred cases of moder-ate and severe OSAHS patients were selected by the method of parallel opening. All of the patients were given health education requirement , quitting smoking and wine , low fat diet and exercise to lose weight and other con-ventional treatment. The patients were randomly divided into the treatment group of 42 cases with noninvasive ventilator treatment , 44 cases treated with conventional treatment , to observe the changes of serum CRP , ET-1 and TNF-α levels and PSG parameters after 12 weeks in two groups. Results Apnea hypopnea index (AHI), oxygen desaturation index (ODI), the lowest oxygen saturation (LSpO2), and the average oxygen saturation MSpO2 in OSAHS patients were significantly improved after treatment (P < 0.01), but the degree of improvement in the two groups after treatment was significantly higher than the control group (P < 0.01). Plasma CRP, ET-1 and TNF-α levels in the two groups after treatment were lower than before treatment (P < 0.05 or P < 0.01), but the treatment group was significantly higher than that of the control group ( P < 0 . 05 or P < 0 . 01 ) . Conclusion Noninvasive ventilator therapy in improving the OSAHS monitoring data of patients with PSG can effectively reduce the serum CRP, TNF-α, ET-1 level, reduce the body′s inflammatory reaction.
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ObjectiveTo compare the effects of two methods for the treatment of delayed defecation of neonatal meconium. Methods Sixty-seven neonates with delayed defecation of neonatal meconium were divided into experiment group (n=37) and control group (n=30). The former group was managed with abdominal massage followed by glycerol enema and the latter with abdominal massage followed by anus stimulation.Then the groups were compared in terms of the time for initial defecation of meconium,the volume of defecation,exhanstion time for defecation,abdominal distention and vomiting and serum bilirubin within 7 days.Result The treatment group was superior to the control one in terms of time for initial defecation of meconium,the volume of defecation, exhanstion time for defecation (P<0.05) and the incidences of abdominal distention,vomiting and serum bilirubin within 7 days were significantly lower than the control group (P<0.05).Conclusions The method of glycerol enema combined with abdominal massage is more effective for promoting defecation of neonatal meconium than the method of anus stimulation.It can reduce the incidence of abdominal distension,vomiting and pathological jaundice.
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<p><b>OBJECTIVE</b>To study the differentially expressed genes in asthenospermia to gain a deeper insight into the molecular mechanisms of the disease.</p><p><b>METHODS</b>We analyzed the differentially expressed genes in asthenospermia using GATHER, PANTHER and ToppGene online bioinformatics tools.</p><p><b>RESULTS</b>Our bioinformatics mining and analyses revealed that the differentially expressed genes in asthenospermia played important roles in the cellular protein and macromolecular metabolism, protein modification, cell death, cell apoptosis and apoptosis induction.</p><p><b>CONCLUSION</b>Asthenospermia patients experience a decline in sperm activity and the basic life activities of sperm simultaneously, and are also prone to cell apoptosis or death. Such differentially expressed genes as KIF3B, MYO15A, KIF6, KIF26B, KIF3A, DNHD2, DMN, DYNC2H1, STARD9, MYOHD1, and TPM1, which are involved in cytoskeletal structure, microtubule movement and cell movement, may be associated with asthenospermia, and therefore deserve further studies.</p>
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Humans , Male , Asthenozoospermia , Genetics , Metabolism , Computational Biology , Databases, Genetic , Gene Expression Profiling , Oligonucleotide Array Sequence Analysis , Spermatozoa , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To investigate the mRNA and protein expression levels of cysteine-rich secretory protein 2 (CRISP2) in the sperm of asthenospermia patients, and explore their relationship with sperm motility and related molecular mechanism.</p><p><b>METHODS</b>We collected 78 semen samples from adult male patients with asthenospermia and another 70 from healthy volunteers as controls. We extracted total RNA and total protein from the sperm following purification of the sperm by Percoll gradient centrifugation, and detected the relative expressions of CRISP2 mRNA and protein in the two groups by RT-PCR, SYBR Green real-time PCR and Western blot.</p><p><b>RESULTS</b>The expression of CRISP2 mRNA was down-regulated by 4.3 times and that of the CRISP2 protein by 1.71 times in the asthenospermia patients, significantly lower than in the normal control group (P < 0.05).</p><p><b>CONCLUSION</b>The down-regulation of CRISP2 mRNA and protein expressions in the sperm of asthenospermia patients may be closely related with decreased sperm motility, which suggests that CRISP2 may serve as a potential molecular target for the research of asthenospermia.</p>
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Adult , Humans , Male , Asthenozoospermia , Genetics , Metabolism , Case-Control Studies , Glycoproteins , Genetics , Metabolism , Sperm Motility , Spermatozoa , Metabolism , PhysiologyABSTRACT
OBJECTIVE@#To determine the correlation of serum soluble CD14 (sCD14) level with the injury of vascular endothelial cells and chronic low grade inflammation in newly diagnosed Type 2 diabetes (T2DM).@*METHODS@#ELISA was used to examine serum sCD14 and serum soluble E-selectin (sE-selectin) level, while immunoturbidimetric assay was used to detect serum high sensitivity C reactive protein (hsCRP).@*RESULTS@#The levels of serum sCD14, sE-selectin, and hsCRP in newly diagnosed T2DM group were higher than those in the euglycemic group [sCD14: (300.7+/-136.6) ng/mL vs. (273.3+/-86.0) ng/mL); sE-selectin: (21.3+/-7.7) ng/mL vs. (32.9+/-11.4) ng/mL; hsCRP: (1.45+/-1.21) mg/L vs. (2.37+/-1.45)mg/L], and there was a significant difference in the latter two parameters between the 2 groups(P<0.01). In the patients with newly diagnosed T2DM, after matching blood pressure, blood sugar, and blood lipid, the levels of serum sCD14, sE-selectin, and hsCRP in the obese group were higher than those in the non-obese group. There was no significant difference in the former 2 parameters between the 2 groups. The serum sE-selectin was correlated with fasting blood sugar (r=0.369, P<0.001), 2-hour postprandial blood sugar (r=0.421, P<0.001), glycosylated hemoglobin (r=0.291, P=0.005), sCD14(r=0.312, P=0.002), and homeostasis model assessment-insulin resistance(r=0.247, P=0.018) in the newly diagnosed T2DM group. Stepwise regression ana-lysis showed that the serum sCD14 was one of the chief influencing factors on serum sE-selectin.@*CONCLUSION@#Serum sCD14 levels tend to increase in newly diagnosed T2DM patients, especially in the obese diabetic patients, which is one of the chief influencing factors to induce the injury of vascular endothelial cells. The innate immunity mediated by Toll-like receptor 4 may take part in the injury of vascular endothelial cells in newly diagnosed T2DM patients.
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Aged , Female , Humans , Male , Middle Aged , C-Reactive Protein , Metabolism , Diabetes Mellitus, Type 2 , Blood , Allergy and Immunology , E-Selectin , Blood , Lipopolysaccharide Receptors , Blood , Regression Analysis , Solubility , Toll-Like Receptor 4 , Allergy and ImmunologyABSTRACT
Objectives To explore the effect of transient continuous subcutaneous insulin infusion (CSII) on β cell function, insulin resistance and vascular endothelial injury in newly diagnosed type 2 diabetic patients and its potential mechanism. Methods Ten patients with newly diagnosed type 2 diabetes mellitus (T2DM) accepted CSII for two weeks. Intravenous glucose tolerance test (IVGTT) and hyperinsulinemia euglycemia clamp test were performed before and after CSII. Serum soluble E-selectin (sE-selectin) was used to evaluate the injury of vascular endothelial cell, while serum high sensitivity C reactive protein (hsCRP) and soluble CD_(14) (sCD_(14)) were both used to assess inflammatory condition. Results (1) Compared with those before treatment, the blood glucose levels of IVGTT, the area under the curve of the blood glucose, glycosylated hemoglobin, TC and LDL-C in the patients were decreased after CSII (P < 0. 05 or 0. 01). (2) Compared with those before treatment, the insulin levels of IVGTT (except the fasting insulin), the area under the curve of insulin and acute insulin response were all increased after CSII(P < 0.05 or 0.01). (3) Compared with that before treatment, the glucose infusion ratio in the clamp test [(3.46±1.66)mg·kg~(-1)·min~(-1) increased to (7.14±2.37)mg·kg~(-1)·min~(-1)]and HOMA-β elevated, while HOMA-IR declined (P <0. 05 or 0. 01 in all). (4) Compared with those before treatment, the levels of serum sE-selectin, sCD_(14) and hsCRP were decreased (P < 0. 01, except for hsCRP) . Conclusion Transient intensive insulin therapy in patients with newly diagnosed T2DM is useful to restore 13 cell function, attenuate insulin resistance, repair vascular endothelial injury and improve the disorder of blood sugar and lipid. The mechanism may be related with the inhibition of inflammation in patients.
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Objective To explore the molecular mechanisms of primary amenorrhea by using arrayCGH technology. Methods Ten patients with primary amenorrhea and 10 female volunteers with regular menstrual cycles as healthy controls were selected. All patients and control samples were analyzed by conventional chromosome analysis (G-banding technology) and array-CGH technology, respectively. ArrayCGH was performed using Affymetrix Cytogenetic 2. 7M arrays following the manufacturer's standard protocol. Results Both the patient group and control group analyzed by conventional G-banding karyotype technology showed a negative result with a normal female karyotype: 46, XX. The result of array-CGH analysis demonstrated a microdeletion of approximately 110 000 bp located at the end of the short arm of X chromosome [46, X, del (X) (p22. 33 )] were identified in 5 patients, which was not detected in the control group. All healthy control samples by array-CGH analysis showed no pathological DNA copy number variation. Conclusions Array-CGH technology can improve the diagnosis rate of chromosomal disease at the DNA level. It is necessary to provide array-CGH for higher resolution genetic analysis of idiopathic primary amenorrhea patient who can not be identified by conventional technology.
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This study was aimed to clarify whether valproic acid (VPA) induces apoptosis of leukemia HL-60 cell line and its possible mechanism. The effect of different concentrations and treatment time of VPA on HL-60 cell proliferation was assayed by cytotoxicity test (CCK-8 method) and fluorescence microscopy, and flow cytometry was used to detect cell apoptosis. The expressions of telomerase subunit h-tert mRNA and apoptosis-related protein as well as caspase-3 activity were detected by real time-quantitative PCR, Western blot and ELISA respectively. The results indicated that VPA inhibited proliferation of HL-60 cells and induced cell apoptosis in a dose dependent manner (r = -0.87). The expressions of anti-apoptotic protein BCL-2 and h-tert mRNA were significantly decreased while the pro-apoptotic protein BAX and caspase-3 activity increased after treatment with VPA. The apoptosis rate of HL-60 cell was negatively correlated with expression of h-tert mRNA. It is concluded that VPA can inhibit leukemia HL-60 cell proliferation and induce apoptosis. The VPA displays anti-leukemia activity possibly through reducing h-tert mRNA and BCL-2 protein expression, increasing BAX expression and activity of caspase-3.
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Humans , Apoptosis , Caspase 3 , Metabolism , HL-60 Cells , Proto-Oncogene Proteins c-bcl-2 , Metabolism , Telomerase , Metabolism , Valproic Acid , Pharmacology , bcl-2-Associated X Protein , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To appraise the effect of single- and two-layer Percoll density gradient centrifugation in sperm separation.</p><p><b>METHODS</b>Twenty semen specimens underwent single-(50%) and two-layer (90% and 45%) density gradient centrifugation, respectively. The sperm class analyzer (SCA) was used to analyze sperm density, motility and dynamic parameters and round cell density before and after the treatment.</p><p><b>RESULTS</b>After separation, the sperm recovery rate of the single-layer method was (65.5 +/- 12.8)%, significantly higher than that of the two-layer method (P < 0.01). The percentages of grade a sperm of the single- and two-layer method were significantly higher than pre-treatment (P < 0.05, P < 0.01), that of the single-layer was significantly lower than that of the two-layer method (P < 0.05), but the percentage of grade c sperm of the former was significantly higher than that of the latter (P < 0.05). Compared with pre-treatment, the percentage of grade a + b sperm of the two-layer method was significantly higher (P < 0.05), while that of the single-layer method showed no significant difference (P > 0.05), and the round cell density of both the methods was significantly lower (P < 0.05, P < 0.01), with no significant differences between the two methods (P > 0.05).</p><p><b>CONCLUSION</b>The single-layer method yields a higher rate of sperm recovery and causes little change in the sperm motility, while the two-layer method effects a lower rate and significantly improves sperm motility. Both the methods can efficiently separate sperm from round cells, and each has its own advantages and its application value in in vitro treatment of sperm.</p>