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Objective To analyze the mutation of common driver genes in patients with non-small cell lung cancer(NSCLC)in eastern Henan Province.Methods A retrospective analysis was performed on 661 patients with NSCLC admitted to the First People's Hospital of Shangqiu city from March 2022 to July 2023.Five kinds of gene mutation detection kits(fluorescent PCR)were used for gene detection in all enrolled patients.The relationship between the clinical features and the status of each driver gene was analyzed by statistical methods.Results In the 661 patients with NSCLC,the mutation rates of EGFR,KRAS,ALK,ROS1,PIK3CA,BRAF,HER2,RET,MET14 and NRAS were 47.35%,9.68%,5.45%,1.82%,2.87%,1.82%,1.21%,0.91%,0.61%and 0%.Mutations in EGFR,ROS1 and HER2 were more likely to occur in women(P<0.05),while KRAS mutations were more common in men(P<0.05).The mutation rates of EGFR,KRAS and ALK in adenocarcinoma was significantly higher than that in squamous cell carcinoma and NSCLC.NOS(P<0.05),and the mutation rate of PIK3CA in NSCLC.NOS was the highest.The mutation rate of KRAS gene in stage Ⅰ+Ⅱ was significantly higher than that in stage Ⅲ+Ⅳ(P<0.05),and there was no significant correlation between other genes and clinical stage.Compared with smokers,the mutation rate of total driver gene was significantly higher in non-smokers(P<0.05).EGFR,ALK,PIK3CA,ROS1,BRAF and HER2 were more common in non-smokers(P<0.05),while KRAS gene was more common in smokers(P<0.05).The mutation rate of 10 driver genes in sediment cell block samples was 78.67%,and the detection rate was significantly higher than that in other types of samples(P<0.05).Conclusion Com-mon driver genes such as EGFR,KRAS and ALK are correlated with gender,pathological type,clinical stage and smoking.Qualified samples of sediment cells have obvious advantages for gene detection and could be widely pro-moted in patients.ARMS-PCR combined detection of 10 genes could be used as the preferred gene detection method for newly diagnosed and treated NSCLC patients.
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Objective:To discuss the molecular association pattern of Shenling Baizhu Powder for type 2 diabetes (T2DM) and ulcerative colitis through network pharmacology method based on the theory of "treating different diseases with same method" in TCM.Methods:The TCMSP and ETCM Chinese medicine chemical composition databases were used to obtain the chemical composition and predict the targets of Shenling Baizhu Powder. The OMIM, GeneCards, DrugBank and TTD disease databases were used to obtain the disease targets of T2DM and ulcerative colitis. The intersection targets of chemical composition of Shenling Baizhu Powder, T2DM and ulcerative colitis were obtained by Bioinformatics platform. Cytoscape 3.8.2 software was used to map the "Chinese materia medica-component-target" network and "effective component- intersection target" network. The GO enrichment analysis and KEGG pathway analysis of the intersection targets were performed with the STRING platform. The intersection target protein-protein interaction network was constructed by STRING database, and core targets were obtained using the CytoNCA plugin of Cytoscape 3.8.2 software. AutodockTools software was applied to validate the molecular docking between the core chemical components and the core targets of Shenling Baizhu Powder.Results:Totally 176 chemical components of Shenling Baizhu Powder, 226 corresponding targets, 11 478 T2DM targets, 4 857 Ulcerative targets of ulcerative colitis, and 162 intersection targets of medicinal chemistry components and diseases were obtained. 1 789 related biological processes, 163 molecular functions, 92 cell constituents, and 192 signaling pathways were obtained by GO enrichment analysis and KEGG pathway analysis. The signaling pathways were mainly about AGE-RAGE, TNF, IL-17,MAPK, PI3K-Akt signaling pathways, etc. The core components of Shenling Baizhu Powder were mainly sitosterol, luteolin, kaempferol, naringenin, beta-carotene, etc. The core targets were EGFR, ALB, IL1B, CASP3, ESR1, VEGFA, PTGS2, TNF, IL6, MYC, AKT 1, JUN, TP53, etc. The core chemical components had tight correlation with the core targets by the molecular docking.Conclusions:By acting on core targets such as TNF, IL1B, IL6, AKT1, VRGFA, PTGS2, MYC, JUN, TP53, and regulating IL-17 signaling pathway, TNF signaling pathway, MAPK signaling pathway, AGE-RAGE signaling pathway, etc., Shenling Baizhu Powder improves tissue inflammation damage, mucosal barrier damage, immune regulation imbalance, intestinal flora dysbiosis, insulin resistance, apoptosis, oxidative stress and other biological processes. Therefore Shenling Baizhu Powder can treat T2DM and ulcerative colitis with the same method.
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ObjectiveTo explore the status of mild cognitive impairment （MCI） among the elderly in Shanghai communities， and to identify the factors associated with MCI. MethodsThe Montreal Cognitive Assessment-Basic （MoCA-B） was used to screen for MCI in elderly adults. Logistic regression analysis was conducted to identify the factors associated with MCI. ResultsAmong 629 participants， 226 （35.90%） were positive for MCI. Older age （OR=1.04， 95%CI： 1.01-1.07）， lower family income （average OR=2.20， 95%CI： 1.01-4.80； poor OR=2.59， 95%CI： 1.03-6.50）， hearing impairment affecting daily life （OR=1.86， 95%CI： 1.19-2.91）， and anxiety symptoms （OR=1.58， 95%CI： 1.02-2.44） were associated with the positive for MCI. Living in central urban area （OR=0.57， 95%CI： 0.37-0.89） and having higher social support （OR=0.97， 95%CI： 0.94-1.00） were protective factors for MCI. ConclusionThe current status of MCI among the elderly in Shanghai communities is not optimistic. It is important and necessary to establish the cognitive-friendly community for the elderly.
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Objective:To investigate the effect of growth hormone replacement therapy(GHRT) on glucose and lipid metabolism in patients with hypopituitarism.Methods:Clinical data of patients with hypopituitarism who received GHRT in Department of Endocrine and Metabolic Diseases, Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine from December 2016 to February 2020 were retrospectively analyzed. The patients were divided into normal glucose regulation(NGR) group and impaired glucose regulation(IGR) group according to their glucose metabolism status before GHRT. The changes of the characteristics of glucose metabolism before and after GHRT were analyzed.Results:A total of 30 patients aged(23.0±5.2) years were included, 23 patients in NGR group and 7 patients in IGR group. After 12 months of GHRT, there were no significant changes in fasting plasma glucose(FPG), 2-hour postprandial plasma glucose(2hPG), and insulin sensitivity index(ISI) in both groups(all P>0.05), while homeostasis model assessment insulin resistance(HOMA-IR) in IGR group was significantly decreased compared with that before GHRT( P<0.05). None of the patients in NGR group progressed to IGR or diabetes mellitus, and none of the 7 patients in the IGR group progressed to diabetes mellitus, while 4 of them recovered from impaired glucose tolerance(IGT) to NGR. Triglyceride, total cholesterol, and low density lipoprotein-cholesterol levels were all significantly decreased in two groups(all P<0.05). Multivariate linear regression analysis showed that the increase of body mass index was an independent risk factor for the increase of FPG and 2hPG( P<0.05). Conclusion:12-month GHRT significantly improved their blood lipid profiles in patients with hypopituitarism without adversely affecting glucose homeostasis.
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@#The CRISPR-Cas (clustered regularly interspaced short palindromic repeats and CRISPR-associated) system is an "adaptive immune system" found in the genomes of bacteria and archaea which is mediated by RNA and resists foreign nucleic acid invasion.Take advantage of specific recognition of target nucleic acid, CRISPR-Cas system can efficiently edit their target site or accurately regulate gene expression, and now have been developed into a powerful tool for gene editing.According to the different compositions of the effector complex, the system has been divided into two categories: class 1 (type I, type IV, and type III) and class 2 (type II, type V, and type VI).Class 2 system, like the CRISPR-Cas9, is widely used in basic research due to the earliest discovery and best research.However, class 1 has not been maturely developed and utilized though it makes up 90% of the entire CRISPR-Cas system.In this essay, the classification of subtype, the assembly of Cascade complex, the cleavage and degradation mechanism of Cas3, and the application in gene editing of class 1 type I CRISPR-Cas system will be discussed and summarized to provide new ideas and methods for further mechanism studying and application of this category.
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Rheumatoid arthritis (RA) is an autoimmune disease and is mainly characterized by abnormal proliferation of fibroblast-like synoviocytes (FLS). The up-regulated cellular membrane expression of G protein coupled receptor kinase 2 (GRK2) of FLS plays a critical role in RA progression, the increase of GRK2 translocation activity promotes dysfunctional prostaglandin E4 receptor (EP4) signaling and FLS abnormal proliferation. Recently, although our group found that paeoniflorin-6'-
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Objective To investigate the pathological mechanism of spinal injury by axial compression experiment on animal spine, so as to provide references for the treatment, prevention and research of spinal injury. Methods The biomechanical study of rabbit spine segments was performed by axial segment compression experiment. The compression process was recorded and strain analysis was performed by digital image correlation (DIC) technology. Results From the top to the bottom of the spine, the ultimate load and bearing capacity of the segment increased continuously; the average limit load of the corresponding single vertebral body was significantly larger than the segment; the strain of the intervertebral disc in the horizontal and vertical directions was significantly larger than that of the upper and lower vertebral bodies. Conclusions In the process of spine compression, the bearing capacity of the intervertebral disc should be taken into account and the injury of spinal segments is mainly manifested as abnormality of the intervertebral disc. The research findings contribute to the prevention and treatment of spinal compression fractures, as well as the design of related therapeutic instruments and assistive devices.
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Acute poisoning is a component of emergency medicine and a key public health problem in clinical toxicology. In recent years, the research and development of industrial chemicals and drugs have developed rapidly, and the incidence of acute drug poisoning has been increasing. It is very important to strengthen the application research of clinical toxicology in acute poisoning, to identify rare and new toxic drugs, and to create conditions for rapid detection of toxic substances. Therefore, this article reviews the types of acute poisoning, the epidemiological characteristics, the detection technology and significance of clinical toxicology, the role of clinical toxicology in the treatment of acute poisoning and its application.
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Objective: To explore the expression of Dickkopf-3(DKK-3) mRNA in oral squamous cell carcinoma(OSCC) and the relationship between the promoter methylation status and the carcinogenesis of OSCC. Methods: The expression of DKK-3 gene in 51 cases of OSCC and corresponding normal mucosa tissue was detected by PT-PCR and methylation-specific PCR, respectively. The relationship between DKK-3 and the clinical pathological features of the patients was analyzed with SPSS 13. 0. Results: The expression level of DKK-3 in OSCC group was lower than that in the control(t =-12. 580, P＜ 0. 05). The methylation rate of DKK-3 gene promoter region in OSCC group was significantly higher than that in the control(χ2 = 19. 273, P＜ 0. 05). The mRNA expression level of DKK-3 gene in OSCC with methylation group was lower than that in the control(t =-2. 817, P＜ 0. 05). Conclusion: Down-regulation of DKK-3 gene expression and hypermethylation of promoter region are important mechanisms in the pathogenesis of OSCC. The hypermethylation of DKK-3 promoter may be the main cause of transcriptional silencing.
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Objective To investigate the relationship between maintaining concentration and loss of consciousness (LOC) concentration of propofol target controlled infusion (TCI) in patients undergoing heart valve replacement.Methods Thirty patients undergoing elective heart valve replacement were enrolled to receive propofol by ladder plasma TCI for anesthesia induction,8 males and 22 females.The initial plasma concentration (Cp) of propofol was set to 1.0 μg/ml,0.3 μg/ml Cp was increased every 1 min until LOC when the prediction effect-cite concentration (Ce) reached 0.5 μg/ml,then sufentanil 0.8-1.0 μg/kg and rocuronium 0.6-0.9 mg/kg were given for intubation.When BIS reached 50,Cp was decreased to the level of Ce.All the surgeries were performed under hypothermia CPB.MAP,HR,CVP,CO,SV,SVR,BIS,propofol Cp and Ce values were recorded at baseline (T0),LOC (T1),BIS reached 50 (T2),and other time points during operation (T3-T9).The correlation analysis between propofol Ce at LOC and perioperative variables were completed.Results In correlation analysis,there was a significant positive correlation between propofol Ce at LOC and baseline CO,SV (P<0.01),there was a significant negative correlation between propofol Ce at LOC and age (P<0.05),there was a significant positive correlation between propofol Ce at LOC and propofol Ce at T2-T9(P<0.01).Conclusion In patients undergoing valvular replacement,the Ce of propofol at maintenance are related to the concentration of propofol at LOC,which is helpful for adjusting the Ce of propofol at maintenance according to the Ce of propofol at LOC.
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To improve the recognition and treatment of the combination of classical congenital adrenal hyperplasia (CAH) and Turner's syndrome. A case of 21-hydroxylase deficiency (21-OHD) in 45,X[3] / 46,XX [47] was reported,and the related literatures were reviewed. A 29-year-old woman with 45,X[3] / 46,XX[47] was referred with clitorimegaly and primary amenorrhea. Her height was 150 cm with a weight of 56 kg. Physical examination revealed a Tanner stage Ⅵ for both breast development and pubic hair development. She showed a little signs of Turner' s syndrome, such as cubitus valgus. Lab findings: sex hormones are significantly increased, including progesterone, testosterone, dehydroepiandrosterone, 17 hydroxyprogesterone, dihydrotestosterone, and androstendione. Enhanced CT scan showed bilateral adrenal hyperplasia. Gynecological ultrasound showed that the size of the uterus and ovary were near normal and the endometrium was not clear. By gene mutation screening, two mutation sites were found in CYP21A2 gene, such as IVS2-13C/ A→G and p. Ile173Asn( c. 518T→A). Taken together, the patient was diagnosed as a combination of 21-OHD and Turner syndrome. A total of ten patients associated with CAH in Turner's syndrome have been reported so far. The findings showed that routine karyotyping during investigations of patients presenting with ambiguous genitalia or with a diagnosis of CAH may reveal the concomitant presence of Turner's syndrome. We should make a definite diagnosis and give early treatment as soon as possible.
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To improve the recognition and treatment of the combination of classical congenital adrenal hyperplasia (CAH) and Turner's syndrome. A case of 21-hydroxylase deficiency (21-OHD) in 45,X[3] / 46,XX [47] was reported,and the related literatures were reviewed. A 29-year-old woman with 45,X[3] / 46,XX[47] was referred with clitorimegaly and primary amenorrhea. Her height was 150 cm with a weight of 56 kg. Physical examination revealed a Tanner stage Ⅵ for both breast development and pubic hair development. She showed a little signs of Turner' s syndrome, such as cubitus valgus. Lab findings: sex hormones are significantly increased, including progesterone, testosterone, dehydroepiandrosterone, 17 hydroxyprogesterone, dihydrotestosterone, and androstendione. Enhanced CT scan showed bilateral adrenal hyperplasia. Gynecological ultrasound showed that the size of the uterus and ovary were near normal and the endometrium was not clear. By gene mutation screening, two mutation sites were found in CYP21A2 gene, such as IVS2-13C/ A→G and p. Ile173Asn( c. 518T→A). Taken together, the patient was diagnosed as a combination of 21-OHD and Turner syndrome. A total of ten patients associated with CAH in Turner's syndrome have been reported so far. The findings showed that routine karyotyping during investigations of patients presenting with ambiguous genitalia or with a diagnosis of CAH may reveal the concomitant presence of Turner's syndrome. We should make a definite diagnosis and give early treatment as soon as possible.
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Objective To explore more suitable calculation method of the insulin dosage in insulin hypoglycemia-growth hormone stimulation test(insulin tolerance test, ITT). Methods Fifty-six subjects suspected of growth hormone deficiency were divided into primary and secondary onset groups. All the patients took oral glucose tolerance test and ITT. Homeostasis model of assessment for insulin resistance index ( HOMA-IR) and insulin sensitivity index ( ISI), area under insulin curve ( AUCINS ) and the area under glucose curve ( AUCPG ) were calculated. The insulin dosages during ITT between two groups were compared and the main factors influencing the insulin dosage were analyzed. Results There was no difference in the insulin dosage during ITT between primary and secondary groups. The actual dosage of insulin in this cohort study revealed a significant difference from the initial insulin dosage recommended by the guideline. Multiple linear regression analysis found that AUCINS and body mass index were the independent factors affecting the insulin dosage. Then the optimized coefficient of ITT ( γ) were found. Conclusion The insulin dosage used in our study was inconsistent with the guidelines-recommended ones. In order to make ITT more efficient and safer, a more optimized calculation method to improve the successful rate of insulin-induced hypoglycemia in ITT is proposed.
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Background: Rice seed proteins are lacking essential amino acids [EAAs]. Genetic engineering offers a fast and sustainable method to solve this problem as it allows the specific expression of heterologous EAA-rich proteins. The use of selectable marker gene is essential for generation of transgenic crops, but might also lead to potential environmental and food safety problems. Therefore, the production of marker-free transgenic crops is becoming an extremely attractive alternative and could contribute to the public acceptance of transgenic crops
Objectives: The present study was conducted to examine whether AmA1 can be expressed specifically in rice seeds, and generate marker-free transgenic rice with improved nutritive value
Materials and Methods: AmA1 was transferred into rice using Agrobacterium-mediated co-transformation system with a twin T-DNA binary vector and its integration in rice genome was confirmed by southern blot. Transcription of AmA1 was analyzed by Real-Time PCR and its expression was verified by western analysis. Protein and amino acid content were measured by the Kjeldahl method and the high-speed amino acid analyzer, respectively
Results: Five selectable marker-free homozygous transgenic lines were obtained from the progeny. The expression of recombinant AmA1 was confirmed by the observation of a 35 kDa band in SDS-PAGE and western blot. Compared to the wild-type control, the total protein contents in the seeds of five homozygous lines were increased by 1.06 tilde 12.87%. In addition, the content of several EAAs, including lysine, threonine, and valine was increased significantly in the best expressing line
Conclusions: The results indicated that the amino acid composition of rice grain could be improved by seed-specific expression of AmA1
الموضوعات
Amaranthus , Agrobacterium , Amino Acids, Essential , Genes, Plantالملخص
Objective To evaluate the relationship between endoplasmic reticulum stress and diabetes mellitus (DM)-caused influence on cardioprotection induced by remifentanil postconditioning in rats.Methods Adult male Sprague-Dawley rats,weighing 250-300 g,were used in the study.A model for type 1 DM was established by intraperitoneal streptozotocin 50 mg/kg and confirmed by blood glucose ≥ 16.7 mmol/L.Thirty six rats with type 1 DM were randomly divided into 3 groups (n =12 each) using a random number table:sham operation group (DM-S group),myocardial ischemia-reperfusion (I/R) group (DM-I/R group) and remifentanil postconditioning group (DM-R group).Another 36 normal rats were exposed to single intraperitoneal injection of sodium citrate-hydrochloric acid buffer solution and served as control group.Two weeks later 36 normal rats with nondiabetes mellitus were also randomly divided into 3 groups (n =12 each) using a random number table:sham operation group (NDM-S group),myocardial I/ R group (NDM-I/R group) and remifentanil postconditioning group (NDM-R group).Myocardial I/R was produced by 30 min occlusion of left anterior descending branch of coronary artery followed by 120 min reperfusion.Remifentanil postconditioning was induced by 10 min infusion of remifentanil 10 μg · kg-1 · min 1 via the femoral vein starting from 5 min before reperfusion.Before ischemia and at 30 and 120 min of ischemia,MAP,SP and HR were recorded and rate-pressure product (RPP) was calculated.At 120 min of reperfusion,arterial blood samples were collected for measurement of plasma cardiac troponin I (cTnI) concentration.The animals were then sacrificed and hearts were removed for determination of myocardial infarct size (IS).The left 6 rats from each group were sacrificed at 120 min of reperfusion,the specimens from their left ventricular apex were obtained to detect the expression of endoplasmic reticulum stress marker glucose-regulated protein 78 (GRP78),C/EBP homologous protein (CHOP) and caspase-12 by Western blot.Results MAP and RPP were significantly decreased,the plasma concentration of cTnI was increased,changes of cardiac infarction were found,and the expression of GRP78,CHOP and caspase-12 was up-regulated in diabetic and nondiabetic rats.Remifentanil postconditioning could inhibit the expression of GRP78,CHOP and caspase-12,increase MAP and RPP,decrease the plasma concentration of cTnI,and reduce myocardial infarct size in nondiabetic rats,but it had no such effects in the diabetic rats.The expression of GRP78,CHOP and caspase-12 was significantly higher after remifentanil postconditioning in diabetic rats than in nondiabetic rats.Conclusion Enhanced endoplasmic reticulum stress is involved in DM-caused loss of cardioprotection induced by remifentanil postconditioning in rats.
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Objective To analyze and study the efficacy of omeprazole and cisapride treatment of reflux esophagitis. Methods From May 2013 to January 2015, 100 cases of patients with reflux exophagitis,were randomly divided in to two groups 50 patients in each group. The control group was treated with omeprazole treatment; and the experimental group on the basis of omeprazole plus cisapride,after the end of the two groups and medication adverse effects,clinical symptom scores were compared. Results The total efficiency of patient test group compared with the reference group were significantly different,statistically significant differences through chi-square test showed (P<0.05). Among them,the experimental group,38 cases markedly effective in 11 cases, one case, the total effective rate was 98.0%. control group 32 cases markedly effective in 12 cases, 6 cases, total effective rate was 88.0%. Adverse drug test group with similar reference group, no significant difference between the two groups were 6.0%, after a chi-square test showed(P>0.05). A test group of clinical symptoms scores compared with the control group,with significant difference,the difference was statistically significant(P<0.05), where the test group was(1.32±0.12)points, the reference group was(2.64±0.15)points. Conclusion Cisapride plus omeprazole treatment of reflux esophagitis significant effect,com-pared with simply using omeprazole treatment,the effect is more precise,will help to improve the clinical symptoms,and safe,worthy promote the use of clinical practice.
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BACKGROUND:Some scholars believe that anesthesia can improve the quality of cord blood separation and colection, and subarachnoid block is commonly used in cesarean section, but there is no research on the influence of subarachnoid block on differentiation potential of mononuclear cels into neural stem cels and astrocytes. OBJECTIVE: To study the influence of subarachnoid block on differentiation potential of mononuclear cels in the cord blood of neonates into neural stem cels and astrocytes. METHODS:Mononuclear cels isolated from 20 neonates delivered spontaneously and 20 neonates born by cesarean delivery undergoing subarachnoid block were culturedin vitro, acting as control group and study group. After delivery, cord blood samples were taken to isolate and culture mononuclear celsin vitro. After 3 days of routine culture, the cels were subject to non-induced culture and induced differentiation into neural stem cels. The cultured cels were identified by the cellmakers Nestin and glial fibrilary acidic protein with immunohistochemistry identification. RESULTS AND CONCLUSION:The Nestin and glial fibrilary acidic protein showed no difference between the two groups after induced differentiation (P > 0.05), indicating subarachnoid block has no impact on the differentiation of mononuclear cels in the cord blood from neonates into neural stem cels and astrocytes.
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Objective To evaluate the effects of a comprehensive intervention including regular aerobic exercise and health education on knowledge,attitude,belief,practice (KAP) scores,body mass index (BMI),waist circumferences,blood pressure,serum glucose and lipid levels in middle-aged and elderly people with overweight and obesity.Methods Three resident communities were chosen randomly from Wangdingdi community in Nankai district in Tianjin.Totally 125 overweight people were involved in this study,62 and 63 people were randomly assigned to intervention group and control group respectively.The members of intervention group were asked to participate in aerobic exercise and health education program 3 times per week within 6 months.And control group members received no intervention.Then to evaluate the effects of intervention of regular aerobic exercise and health education by physical examination and questionnaire.Comparison between intervention group and control group was performed using a repeatedmeasures analysis of variance.Results The comprehensive intervention resulted in significant improvement of the KAP scores [13.82 (95％CI:12.42-15.23)] and high-density lipoprotein cholesterol [0.22(95％ CI:0.17-0.28) mmol/L] and reductions in intervention group in body mass index [-0.47 (95％ CI:-0.58-0.36) kg/m2],waist circumference [-4.00 (95％ CI:-5.01--2.99) cm],systolic blood pressure [-11.69 (95％ CI:-13.84--9.55) mmHg] (1 mmHg=0.133 kPa),fasting plasma glucose [-1.21 (95％ CI:-1.55--0.88) mmol/L],triglyceride [-0.06 (95％ CI:-0.25-0.13) mmol/L],low-density lipoprotein cholesterol [-0.48 (95％ CI:-0.62--0.34) mmol/L].Conclusion Regular aerobic exercise and health education program in community can successfully reduce body mass index,waist circumference and some other risk factors related to overweight and obesity.It is effective to improve awareness,physical fitness,change lifestyle for middle-aged and elderly people in the community.
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Objective To purify recombinant methioninase and investigate the synergetic inhibitory effect of methioninase and cisplatin on the proliferation of human lung adenocarcinoma cell line GLC. Methods Recombinant methioninase was purified with GST-column from supernatant after ultrasonic disruption of cultured Escherichia coli in the prokaryotic expression system pGEX-4T-1-Met/Dh5a. MTT assay was used to determine the inhibition rate of methioninase in combined with cisplatin on cell proliferation,and their synergistic effect was evaluated by using the q value judge method. Results The concentration of recombinant methionine cleaving enzyme was 0.22 mg/mL, the purity was 95%, and the activity was 0.568 IU/mg. After 72 h culturing, the inhibition rate of cisplatin methionine was 24.80%and 27.49%respectively,while the inhibition rate of the combined drugs was 66.80% ( =1.47>1.15) which showed a significant synergistic effect. Conclusion Both methioninase and cisplatin have the inhibition effect,and the combined drugs display a significant synergistic effect on the proliferation of GLC.
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To improve the extraction of solanesol from tobacco waste, we developed an enzymatic cell wall-breaking process with combined cellulase and ligninase. The effects of reaction time, temperature, pH and enzyme/substrate ratio were determined. The results show that the catalytic effect was better than either single enzyme when the ratio of cellulase to ligninase was 15:1 (U/U). Under the optimized conditions of 175 U/g (enzymes/substrate), tobacco to water 1:5 (W/W), temperature 40 degrees C and pH 6.0, the concentration of solanesol in the solution could reach 0.33 g/L after 8 h. And the average leaching rate reached 96.53% which was 1.68 times of the extraction methods of chemical reflux. It provides new way for the extraction of solanesol from tobacco waste, and worthwhile to be further explored.