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OBJECTIVES@#Obstructive sleep apnea hypopnea syndrome (OSAHS) is a common disease that seriously affects health. Continuous positive airway pressure (CPAP) therapy is the preferred treatment for moderate-to-severe OSAHS patients. However, poor adherence to CPAP is a major obstacle in the treatment of OSAHS. Information-motivation-behavioral (IMB) skills, as a kind of mature technology to change the behavior, has been used in various health areas to improve treatment adherence. This study aims to explore the effects of the IMB skills intervention on CPAP adherence in OSAHS patients.@*METHODS@#Patients who were primary diagnosed with moderate-to-severe OSAHS were randomly divided into the IMB group (n=62) and the control group (n=58). The patients in the IMB group received CPAP therapy and the IMB skills intervention for 4 weeks. The patients in the control group received CPAP therapy and a usual health care provided by a registered nurse. We collected the baseline data of the general information, including age, sex, body mass index (BMI), the Epworth Sleepiness Scale (ESS) score, the Hospital Anxiety and Depression Scale (HADS) score, and indicators about disease severity [apnea-hypopnea index (AHI), percentage of time with arterial oxygen saturation SaO2<90% (T90), average SaO2, lowest SaO2, arousal index]. After CPAP titration, we collected CPAP therapy-relevant parameters (optimal pressure, maximum leakage, average leakage, 95% leakage, and residual AHI), score of satisfaction and acceptance of CPAP therapy, and score of willingness to continue CPAP therapy. After 4 weeks treatment, we collected the ESS score, HADS score, CPAP therapy-relevant parameters, effective CPAP therapy time per night, CPAP therapy days within 4 weeks, CPAP adherence rate, score of satisfaction and acceptance of CPAP therapy, and score of willingness to continue CPAP therapy. Visual analog scale (VAS) of 0-5 was used to evaluate the satisfaction and acceptance of IMB intervention measures in the IMB group.@*RESULTS@#There were no significant differences in the baseline level of demographic parameters, ESS score, HADS score, disease severity, and CPAP therapy related parameters between the IMB group and the control group (all P>0.05). There were no significant differences in score of willingness to continue CPAP therapy, as well as score of satisfaction and acceptance of CPAP therapy after CPAP titration between the IMB group and the control group (both P>0.05). After 4 weeks treatment, the ESS score, HADS score, maximum leakage, average leakage, and 95% leakage of the IMB group were significantly decreased, while the score of satisfaction and acceptance of CPAP therapy and willingness to continue CPAP therapy of the IMB group were significantly increased (all P<0.05); while the above indexes in the control group were not different before and after 4 weeks treatment (all P<0.05). Compared with the control group, the ESS score, HADS score, maximum leakage, average leakage, and 95% leakage of the IMB group after 4 weeks treatment were significantly lower (all P<0.05); the effective CPAP therapy time, CPAP therapy days within 4 weeks, score of satisfaction and acceptance of CPAP therapy, score of willingness to continue CPAP therapy of the IMB group were significantly higher (all P<0.05). The rate of CPAP therapy adherence in 4 weeks of the IMB group was significantly higher than that of the control group (90.3% vs 62.1%, P<0.05). The VAS of overall satisfaction with IMB skills intervention measures was 4.46±0.35.@*CONCLUSIONS@#IMB skills intervention measures can effectively improve the adherence of CPAP therapy in OSAHS patients, and is suitable for clinical promotion.
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Humains , Ventilation en pression positive continue , Motivation , Oxymétrie , Observance par le patient , Syndrome d'apnées obstructives du sommeil/diagnostic , SyndromeRésumé
【Objective】 To establish a dry fluorescent luminescence method for the detection of antibodies to hepatitis C virus (HCV) and evaluate its clinical application. 【Methods】 Anti-HCV antibody was detected by double-antigen sandwich dry fluorescent luminescence method established using multi-epitope chimeric antigen. The established method was used to detect national reference samples(positive 20, negative 20), and a total of 349 clinical samples, including 108 HCV patients, 36 patients with other diseases and 205 healthy individuals, which were tested in parallel with enzyme-linked immunoassay (ELISA) to evaluate the performance of the established method. 【Results】 The concordance rate of positive and negative(each 20) reference samples were both 100% (20/20), and the CV of precision reference sample was 9.16%, which met the requirements of national reference samples. In clinical performance evaluation, the AUC value was 0.984, and the sensitivity and specificity of the dry fluorescent luminescence method were 96.30% (104/108) and 96.27% (233/241). The overall concordance rate between dry fluorescent luminescence method and ELISA was 97.71% (341/349) (Kappa=0.952). 【Conclusion】 The dry fluorescence luminescence method of HCV antibody is simple and rapid, with high sensitivity and high specificity, and can be used in clinical application.
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Objective:This study is to investigate the predictive value of serum levels of TIMP-2 and insulin-like growth factor-binding protein 7(IGFBP7) in patients with DCD(donation after cardiac death) kidney transplantation.Methods:A prospective research design was used to select DCD kidney transplant patients admitted to the Li Huili Hospital of Ningbo University from January 2018 to October 2020.Inclusion criteria: ①Complete data; ②There were no serious complications affecting the function of the transplanted kidney in the early postoperative period.Exclusion criteria: ①Incomplete data; ②Patients were unable or unwilling to cooperate with the study; ③Severe complications affecting the function of the transplanted kidney occurred early after the operation.The ELASE method was used to quantitatively detect the serum TIMP-2 and IGFBP7 levels at 6, 12, 24, 48, 72 hours and 7 days after renal transplantation, and monitor the serum creatinine values during the same period and 21 days after the operation. According to the occurrence of DGF, the measured values of TIMP-2 and IGFBP7 at different time points and their product's ability to predict the occurrence of DGF after kidney transplantation were analyzed. The receiver operating characteristic (ROC) curve and area under the curve (AUC) were used to evaluate the diagnostic efficacy of TIMP-2 and IGFBP7 for DGF.Results:A total of 33 patients were enrolled, 7 patients (21.2%) in the DGF group and 26 patients (78.8%) in the non-DGF group. Between the two groups, the donor glomerular filtration rate were [98.5(15.8-132.5)ml/(min·1.73m 2) and 79.1(60.6-102.5)ml/(min·1.73m 2)], recipient gender (male/female: 3/4 cases and 10/16 cases), recipient age [48(34-56) Years old and 45(23-61) years old], the recipient's preoperative creatinine [1114.0(731.4-1293.0)μmol/L and 858.4(657.6-1051.9)μmol/L], the recipient's preoperative urea nitrogen [15.0(13.2-19.6)mmol/L and 17.3(13.6-20.9)mmol/L], receptor preoperative albumin [43.5(38.5-45.3)mmol/L and 41.2(37.5-46.1) mmol/L], recipient dialysis method [hemodialysis/peritoneal dialysis: 3/4 cases and 9/17 cases], warm ischemia time [6(5-7) and 5(4-6) min, there was no statistically significant difference] ( P>0.05). The values of serum IGFBP7 and TIMP-2×IGFBP7 in the DGF group were higher than those in the non-DGF group at all time points ( F=15.753, P=0.040; F=13.000, P=0.024), while serum TIMP-2 was not significant between the two groups difference ( F=1.157, P=0.075). For the diagnostic value of DGF, the AUC of serum IGFBP7 at 48 h after surgery was 0.863 (95% CI 0.696-1.000, P=0.004). When 5.97 ng/ml was used as the cut-off value, the sensitivity was 85.7% and the specificity was 80.8 %. The AUC of TIMP-2×IGFBP7 at 48 hours after surgery was 0.819 (95% CI 0.641-0.996, P=0.011). When 62.06(ng/ml) 2 was used as the cutoff value, the sensitivity was 71.4% and the specificity was 80.8%.There was no statistical difference in the area under the curve between the two ( P>0.05). There were differences in the dynamic trend of serum IGFBP7 and creatinine in the DGF group. Serum IGFBP7 at 7 days after surgery was positively correlated with creatinine at 21 days after surgery. Conclusion:Serum IGFBP7 and TIMP-2×IGFBP7 could predict the occurrence of DGF after DCD donor kidney surgery. The predictive value changes with time. Among them, 48h and 7d after surgery are the most valuable. However, serum TIMP-2 has not been found to have predictive value in this study.
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Objective:To explore the predictive values of tissue inhibitor of metalloproteinase-2 (TIMP-2) and insulin-like growth factor binding protein 7 (IGFBP7) in donor sera and lavage fluid on delayed graft function (DGF) in donation after circulatory death (DCD) kidney transplant recipients.Methods:A total of 33 eligible kidney donors and 33 corresponding recipients were recruited. Preoperative serum and renal perfusion fluid samples of donors were collected to determine the levels of TIMP-2 and IGFBP7. Patients were grouped according to whether DGF occurred after kidney transplantation and measured indicators analyzed. Independent sample t test was utilized for comparing the groups with normal distribution measurement data. And χ2 test was employed for comparing the groups with normal distribution counting data and Mann-Whitney test for comparing the groups with non-normal distribution measurement data. Receiver operating characteristic (ROC) curve and area under curve (AUC) were used for evaluating the diagnostic efficacy of indicators. Results:In donor-DGF group, lavage fluid TIMP-2, product of lavage fluid TIMP-2 and IGFBP7 (TIMP-2×IGFBP7), serum IGFBP7 and product of serum TIMP-2 and IGFBP7 (TIMP-2×IGFBP7) were higher than those in donor-non-DGF group ( P<0.05). The AUC of TIMP-2, TIMP-2×IGFBP7, serum IGFBP7 and serum TIMP-2×IGFBP7 in the diagnosis of DGF were 0.753 (95%CI 0.546~0.959), 0.747 (95%CI 0.510~0.984), 0.824 (95%CI 0.615~1.000) and 0.852 (95%CI 0.660~1.000) respectively. Conclusions:Donor serum IGFBP7, donor serum TIMP-2×IGFBP7, lavage fluid TIMP-2 and lavage fluid TIMP-2×IGFBP7 may be used for predicting the occurrence of early DGF after kidney transplantation. Among them, serum TIMP-2×IGFBP7 has the highest diagnostic efficiency and may be an excellent predictor of DGF occurrence.
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Many countries in the world have faced with coronavirus disease 2019 (COVID-19) epidemic since December 2019, while the proportion of elderly patients with COVID-19 in severe and death cases is relatively high. At present, China is in the rapid development stage of population aging, and the demand of the elderly for medical care, health care, nursing and life services far exceeds that of other people. Especially in the period of COVID-19, it is particularly urgent to summarize more experience and methods in time to reduce the infection rate, the incidence of critical illness, and the mortality rate. Therefore, this review combines the existing research results with clinic experience of diagnosis and treatment for senile infectious diseases, summarizes the clinical characteristics and puts forward the prevention strategies of elderly COVID-19 patients, which provide evidence for effective prevention and treatment of COVID-19 in elderly patients, improvement of cure rate, and reduction of severe incidence rate and mortality.
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Sujet âgé , Humains , Betacoronavirus , Chine , Infections à coronavirus , Diagnostic , Mortalité , Thérapeutique , Pandémies , Pneumopathie virale , Diagnostic , Mortalité , ThérapeutiqueRésumé
Objective:To analyze the antibody level of phospholipase A2 receptor (PLA2R) in Chinese patients with primary membrane nephropathy (PMN) and its correlation with clinical indicators, and to explore more suitable cut-off value for Chinese patients.Methods:All hospitalized patients with renal biopsy at Peking University People's Hospital from January to August 2018 were retrospectively reviewed. According to the primary disease, patients were divided into PMN group (including patients with idiopathic membranous nephropathy and atypical membranous nephropathy of unknown cause) and control group (other pathological types, including secondary membranous nephropathy patients). Their clinical and pathological characteristics were analyzed, and the level of serum PLA2R antibodies was detected using the method of enzyme-linked immunosorbent assay (ELISA). Spearman correlation was used to analyze the correlation between PMN patients' blood anti-PLA2R antibody level and clinical indicators. The risk factors of PMN were analyzed by logistic regression model, and the optimal cut-off value of PMN was analyzed by ROC curve.Results:A total of 354 patients were included in this study, including 114 patients in PMN group and 240 patients in control group. The age of PMN group was (51.7±14.1) years old and the ratio of male to female was 2.2∶1. The median concentration of PLA2R antibody in PMN group was 16.87 RU/ml [inter-quartile range ( IQR) 1.88-57.26], which was significantly higher than that in control group (1.43 RU/ml, IQR 1.20-1.62, P<0.001). In PMN group, the concentration of anti-PLA2R antibody was correlated with the 24-hour urine protein ration ( r=0.278, P=0.003) and urine erythrocyte ( r=0.190, P=0.043), but not with serum albumin ( r=-0.149, P=0.114) and serum creatinine ( r=0.136, P=0.149). The ROC curve showed that the sensitivity of distinguishing PMN from other diseases was 69.3% (95% CI 60.3%-77.0%), the specificity was 92.9%(95% CI 89.0%-95.5%), and the area under the curve was 0.859(95% CI 0.813-0.905) when the cut-off value was set as 2.28 RU/ml, which was significantly better than the cut-off value of 20.00 RU/ml (the sensitivity/specificity was 46.5%/97.5%) and 14.00 RU/ml (the sensitivity/specificity was 53.5%/97.1%). Conclusions:PLA2R antibody is one of the main pathogenic antibodies of PMN. In China, it is recommended to lower its cut-off value to 2.28 RU/ml, which can improve the sensitivity of distinguishing PMN from other pathological types without reducing specificity.
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Objective To investigate the effect of chronic intermittent hypoxia(CIH)on serum high-density lipoprotein cholesterol(HDL-C)and the changes of HDL-C metabolism-related indicators such as the expressions of peroxisome proliferators-activated receptor a (PPARα)and adenosine triphosphate binding cassette transporterA1(ABCA1)in liver in male SD rats.Methods Obstructive sleep apnea syndrome(OSAHS)-induced CIH rats were randomly allocated into 6 groups:10%CIH-3 weeks,5%CIH-3 weeks,5%CIH-3 weeks +RH(Removal of hypoxia-3 weeks),10%CIH-3 weeks + RH(Removal of hypoxia-3 weeks),control group-3 weeks,and control group-6 weeks.Serum lipids were measured and compared.To observe and compare the liver pathology,the expression levels of PPARα and ABCA1 in liver tissue of CIH rats were detected by immunohistochemical method.Results The levels of serum TC,TG and LDL-C was significantly higher in CIH rats than in control group.The levels of TC,TG and LDL-C were significantly lower in reoxygenation groups than in CIH groups.There was no significant difference between experiment groups and correspondent control groups(all P>0.05).Compared with control group,CIH rats had significantly lower levels of serum HDL-C;Compared with CIH groups,the levels of HDL-C were significantly higher in reoxygenation groups.There was no significant difference between experiment groups and correspondent control groups(all P>0.05).Compared with control group,the expression of PPARα and ABCA1 of CIH group was significantly lower;Compared with CIH groups,the expression of PPARα and ABCA1 was significantly higher in reoxygenation groups;There was no significant difference in the expression of PPARα and ABCA1 between reoxygenation groups and correspondent control groups,in which the oxygen level was recovered to normal.Conclusions The serum HDL-C level was obviously decreased in OSAHS-induced CIH rats,and the decreased serum HDL-C can be effectively improved by reoxygenation intervention.OSAHS-induced CIH may lead to the dyslipidemia through PPARα-ABCA1 pathway,and reoxygenation intervention for three weeks can effectively recover the expression levels of PPARα and ABCA1 to normal levels,which suggests that if CIH is effectively intervented,the change of PPARα and ABCA1 of liver can be reversed,thereby reversing dvslipidemia.
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Objective To establish and characterize human colorectal cancer orthotopic implantation model in nude mice.Methods Established the human colorectal cancer orthotopic implantation model by surgery,characterize its biological characteristics by immunohistochemistry.Results Human colorectal cancer orthotopic implantation model was established in nude mice successfully,the proliferating cell nuclear antigen and Survivin were overexpressed in the models which were simulated the characteristics of human colorectal cancer perfectly.Conclusion This human colorectal cancer orthotopic implantaion model will provide an excellent animal model for us to study the development and metastasis mechanism of human colorectal cancer.
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Objective To evaluate the clinical characteristics of patients with adrenocorticotropin-independent macronodular adrenal hyperplasia (AIMAH).Methods A total of 10 AIMAH cases were enrolled in this retrospective study.The clinical and laboratory findings of all patients were collected and analyzed.Results All patients manifested some clinical features and biochemical evidence of Cushing's syndrome.The plasma adrenocorticotropic hormone (ACTH) level was undetectable in all the patients and their serum cortisol secretion rhythm was abnormal.Low and high-dose dexamethasone suppression tests failed to suppress the cortisol secretion.The bilateral macronodular adrenal enlargement was shown by CT/magnetic resonance imaging.The supine-upright posture test was positive in four patients.Three patients were performed bilateral adrenalectomy,five were unilateral adrenalectomy and the remaining two patients were taken propranolol.All the patients had followed up for 10 to 89 months.Contralateral adrenalectomy was performed in two patients with recurrent symptoms after unilateral adrenalectomy and two patients given propranolol were underwent bilateral adrenalectomy when their symptoms had not been improved or recurred.Conclusion AIMAH is a relatively rare subtype of Cushing's syndrome with unique clinical and laboratory findings.Propranolol is a good choice if the supine-upright posture test is positive.Unilateral adrenalectomy appears to be an effective and safe alternative treatment for AIMAH.Bilateral adrenalectomy could be performed if the symptoms have not been improved or recurred after unilateral adrenalectomy.
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Objective: To investigate the effect of chronic intermittent hypoxia (CIH) on liver injury and the expression of fractalkine in rats and explore its possible mechanism. Methods: A CIH murine model was established to mimic the pathophysiology of obstructive sleep apnea-hypopnea syndrome (OSAHS) in humans. Thirty healthy male Spraque-Dawley rats were randomly assigned to 3 groups: a 5% CIH group, a 5% CIH+RH (removal of hypoxia) group and a control group ( 10 rats in each group). The 5% CIH and 5% CIH+RH groups were exposed to CIH for 3 weeks, 8 h/d, and the frequency of hypoxia was 20 times/h. The 5% CIH+RH group was then exposed to normal gaseous environment for another 3 weeks. After the experiment, liver sections were stained with hematoxylin-eosin (HE) and the liver pathology was observed. The expression of fractalkine in the liver tissues was detected by immunohistochemical method. Results: 1) Compared with the control group, the hepatic steatosis and inflammatory activities in the 5% CIH and 5% CIH+RH groups were more severe (allP Conclusion: CIH can induce liver injury and high fractalkine expression in rat liver tissues.
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OBJECTIVE@#To investigate the effect of nasal intermittent positive pressure ventilation (NIPPV) on N-terminal pro-brain natriuretic peptide (NT-proBNP) in patients with acute exacerbations of chronic obstructive pulmonary disease (AECOPD) and type II respiratory failure.@*METHODS@#Forty patients with AECOPD and type II respiratory failure and 40 patients with stable phase chronic obstructive pulmonary disease were randomly assigned into study. Plasma levels of NT-proBNP, arterial blood gas, APACHE II scores, and pulmonary artery pressures were measured. The plasma level of NT-proBNP was compared between the two groups. Effect of NIPPV on NT-proBNP was studied in patients with AECOPD and type II respiratory failure.@*RESULTS@#There were negative correlations between NT-proBNP and pH, and between NT-proBNP and PaO2 (r=-0.691,r=-0.704,respectively;P<0.001),positive correlations between NT-proBNP and PaCO2, and between NT-proBNP and APACHE II scores (r=0.774, r=0.810, respectively, P< 0.001), and positive correlation between NT-proBNP and PAP (r=0.965, P<0.001) in all patients. In patients with AECOPD and type II respiratory failure, there were negative correlations between NT-proBNP and pH,and between NT-proBNP and PaO2 (r=-0.636, r=-0.616,respectively; P<0.001); there were positive correlations between NT-proBNP and PaCO2, and between NTproBNP and APACHE II scores (r=0.545, r=0.475, respectively; P=0.001, P=0.002); and there were positive correlation between NT-proBNP and pulmonary artery pressure (r=0.833,P<0.001). The plasma levels of NT-proBNP were significantly higher in patients with AECOPD and type II respiratory failure than in control subjects [(939.60 ± 250.00) pg/mL vs (151.55 ± 111.20) pg/mL;P<0.01]. NIPPV decreased plasma levels of NT-proBNP [(229.15 ± 98.26) pg/mL vs (939.60 ± 250.00) pg/mL; P<0.01] in patients with AECOPD and type II respiratory failure, as well as improved arterial blood gas and APACHE II scores. Although NIPPV appeared to decrease pulmonary artery pressure somewhat between pre-treatment and post-treatment groups, the differences were not statistically significant (P=0.056).@*CONCLUSION@#The plasma level of NT-proBNP reflects the severity of patients with AECOPD and type II respiratory failure. NIPPV can decrease a patient's splasma level of NT-proBNP, which has clinical value for evaluating the effect of NIPPV.
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Sujet âgé , Femelle , Humains , Mâle , Adulte d'âge moyen , Gazométrie sanguine , Peptide natriurétique cérébral , Sang , Fragments peptidiques , Sang , Ventilation à pression positive , Méthodes , Broncho-pneumopathie chronique obstructive , Sang , Thérapeutique , Insuffisance respiratoire , Sang , ThérapeutiqueRésumé
Objective To screen and diagnose obstructive sleep apnea-hypopnea syndrome (OSAHS) in elderly males by obesity index using receiver operating characteristic(ROC) curves. Methods Data of 402 consecutive elderly male patients who underwent polysomnography from 2001 to 2008 were collected. The relationship between apnea hypopnea index(AHI) and obese indexes such as body mass index (BMI), neck circumference (NC), waist circumference (WC) and waist-to-hip ratio (WHR) were analyzed by Pearson's correlation. ROC curves were used to determine the best cutoff values to screen and diagnose OSAHS, and their priority was compared by area under curve (AUC). A two-tailed P value less than 0.05 was considered statistically significant. Statistical analysis was carried out with SPSS version 13.0. Results (1) AHI was positively correlated with BMI (r=0.241,P<0.001), NC(r=0.201,P<0.001), WC(r=0.210,P<0.001) and WHR(r=0. 097,P>0.05)) in elderly male patients. The area under curve (AUC) of BMI, NC, WC and WHR was 0.61, 0.58, 0.51 and 0.45 respectively, and P value was 0.001,0.060,0.840 and 0. 250 respectively. Only BMI was competent in screening and diagnosing OSAHS in elderly male adults; (2) The optimal value of BMI was 22.0 kg/m~2 in screening OSAHS with specificity 90% and rate of missed diagnosis 10%; (3) The optimal value of BMI was 29.0 kg/m~2 in diagnosing OSAHS with specificity 90% and rate of missed diagnosis 10%. Conclusions BMI more than 22.0 kg/m~2 could be the reference standard to screen OSAHS and BMI more than 29.0 kg/m~2 to diagnose OSAHS in elderly men.
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Objective To analyze the association between mtDNA mutations and photodamagc after ultraviolet B (UVB) irradiation. Methods Primary human skin fibroblasts (HSF) and primary human epi- dermal keratinocytes of adult (HEKa) were irradiated by sub-lethal doses of UVB thrice a day for 4-5 days. Thereafter, genomic DNA was extracted from irradiated cells and conventional PCR was applied to detect the frequency rates of 4977 bp and 3895 bp mtDNA deletion. To quantitatively analyze the mutation levels, SYBR Green real-time PCR method was performed. Results In both cell lines, the frequency rates and relative copy number of deletions increased with the cumulative doses of UVB exposure (P<0.05). The prevalence rate of 3895 bp deletion peaked 53.3% and and relative copy number reached (49.63±4.38)×10-5, showing a more intense response to the accumulation of UVB radiation than 4977 bp deletion. In HSF, the minimum cumu- lative dose of UVB radiation was 150 mJ/cm2 for the induction of 3895 bp deletion, and 200 mJ/cm2 for the induction of 4977 bp deletion. It seemed that mtDNA deletion was more readily to be induced by UVB radia- tion in HSF than in HEKa. Conclusions The development and accumulation of mtDNA mutation are intimately related with cumulated UVB dose received by skin cells, and the 3895 bp deletion is more reliable in moni- toring the photodamage caused by UV than 4977 bp deletion. Therefore, the 3895 bp deletion may serve as a biomarker for the detection of photodamagc in skin cells. HSF appear to have an increased susceptibility to UVB radiation, which results in a higher frequency and level of mtDNA mutations compared with HEKa.
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Objective To quantitatively observe the actual levels of 4977 bp and 3895 bp mitochon-drial DNA (mtDNA) deletion in healthy human skin of different age, and to explore their relationship with intrinsic aging and photoaging. Methods Seventy-one samples of skin tissue were obtained from healthy volunteers, including 40 samples from UV-protected areas (buttock, thigh, waist or abdomen) and 31 from UV-exposed areas (neck, back of hands, forehead or face). Nuclear and mitochundrial DNA was extracted from these samples. Conventional PCR was performed to detect the incidence of 4977 bp and 3895 bp mtDNA deletion. Then, SYBR Green real-time PCR was applied to quantitatively analyze the target mutations in posi-tive samples. Results Conventional PCR showed that the incidence of both 4977 bp deletion and 3895 bp deletion increased with age. For example, the incidence of 4977 bp deletion and 3895 bp deletion accounted for 47.5% (19/40) and 30% (12/40), respectively in samples from volunteers older than 40 years, signifi-cantly higher than that in those from volunteers younger than 40 years (X2 = 4.673, 6.118, respectively, both P < 0.05). The total incidence of 4977 bp deletion and 3895 bp deletion in UV-exposed areas was 48.4% (15/31) and 32.3% (9/31), respectively, which did not differ from those in UV-protected areas. The results from real-time PCR revealed a positive correlation of the copy number of 4977 10p deletion and 3895 bp deletion with age (rg = 0.907, 0.845, respectively, both P < 0.05). When the UV-exposed area was compared with the UV-protected area, no significant difference was found in the copy number of 4977 bp deletion ( P = 0.264), whereas a higher level of 3895 bp deletion was noticed in UV-exposed area (P = 0.014). Conclu-sions The 4977 bp mtDNA deletion is primarily associated with chronological aging, and might serve as a biomarker for the process of chronological aging of skin. Deletion of 3895 bp mtDNA seems to be more sus-ceptible to ultraviolet radiation.
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Objective To observe the aging,apoptosis,cell cycle arrest and oxidative stress in human skin fibroblast(HSF)induced by UVB,and to detect the expression profiles of p66Shc,a determinant of oxidative stress response and life span,in this process.Methods HSF cells were exposed to UVB at a subcytotoxic dosage twice a day for three days.The cells without exposure served as control.After another 24-hour culture,SA-β-Gal staining was performed to evaluate the senescence state of the cells,flow cytometry to observe cell apoptosis;cell cycle arrest was detected by serum starvation and flow cytometry:ELISA was applied to detect intracellular levels of superoxide dismutase(SOD)and malondialdehvde(MDA),and Western blotting to analyze the expression of p66Shc protein.Results The percentage of cells positive for SA-β-Gal staining increased from 0 to 98.3% after UVB radiation,which strongly suggested an aging state of HSF cells.The percentage of apoptotic cells increased from 0.96% to 37%.and 80.07% of the HSF cells were arrested in G0/G1 phase following the irradiation.Intracellular SOD activity decreased from(52.35±4.97)ng/g to(7.81±0.68)ng/g(P<0.01).while intracellular MDA was found to increase from(3.52±0.34)ng/g to(33.91±3.20)ng/g(P<0.05).The p66Shc protein was found to be weakly expressed in HSF in 24 hours following the exposure to UVB,and a stronger expression was noted 48 hours later.Conclusions HSF cells are induced into a state of senescence associated with oxidative stress after UVB irradiation,which may be applied as an in vitro model in aging research.The expression of p66Shc is increased in HSF during this process,and further studies are needed to explore the relation between p66Shc and oxidative stress as well as cellular aging.
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The effects of different carbon sources added to aerobic medium on cell growth,enzyme activity and metabolite distribution was investigated in dual-phase fermentations.The results showed that both cell density and enzyme activity were enhanced by adding 4mmol/L glucose or 12,54,80mmol/L sodium acetate to aerobic medium,respectively.Then centrifugated cells that had grown aerobically in different conditions were transferred to anaerobic fermentation,the enzyme activity and metabolite distribution changed.To analyze the anaerobic enzyme activity and metabolite distribution,it was concluded that during the anaerobic fermentation of Escherichia coli NZN111 overexpressed malic enzyme,PEP carboxykinase(PCK)was the key enzyme of succinate production,pyruvate kinase(PYK)was associated to the accumulation of by-product pyruvate,and isocitrate lyase(ICL)have some influence on the concentration of succinate.The yield of succinate in anaerobic stage could reach 89.0%,which was 16.6% higher than that of the control by adding 80mmol/L sodium acetate as carbon source in aerobic medium.
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Objective To observe the therapeutic effects of paclitaxel solution on psoriasis-like pathological changes of guinea pig.Methods5%propranolol ointment was applied on Guinea pigs' ears to produce psoriasis-like pathological changes,then solutions with a paclitaxel concentration of200?g/mL and500?g/mL were applied on the affected ears.The pathological manifestations were investigated under light microscope.Results There were hyperkeratosis,dyskeratosis,acanthosis,psoriasiform elongation of the rete ridges and significant infiltration of mononuclear cells and polymorphic nucleus leucocytes in dermis in the guinea pig models,with the Baker score reaching6.37?0.99averagely.After being treated with paclitaxel solutions of a concentration of200?g/mL and500?g/mL,the pathological manifestations were remarkably improved,with the Baker score reducing to3.13?1.13and2.13?0.81respectively.A significant differ-ence(P