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To explore the effect of icaritin on preventing the vascular calcification in mouse induced by vitamin D2. Methods: Fifty male C57BL mice were randomly assigned to a control group (n=10) and a model group (n=40). Mice in model group were treated with gradient-concentration of icaritin for 8 weeks. Then, they were consecutively treated with vitamin D2 for 4 days. Meanwhile, mice in negative control group were treated with the same dosage of PBS. At the end of the treatment, aortae were collected to examine the concentration of Ca, the area of calcification and the expression of α-smooth muscle actin (α-SMA) and Runx2. Results: Compared with the negative control group, the weight of mice and the concentration of Ca in the positive control (vitamin D2+0 mg/kg icaritin) group declined significantly (P<0.05) after injection of vitamin D2. Compared with the positive control group, the Ca concentration, the area of calcification, and the expression of Runx2 were significantly declined at the middle dosage of icaritin (vitamin D2+0.2 mg/kg icaritin) group (P<0.05). Conclusion: Icaritin could effectively prevent the vascular calcification in mice.
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Animals , Male , Mice , Aorta , Flavonoids , Mice, Inbred C57BL , Muscle, Smooth, Vascular , Vascular CalcificationABSTRACT
Objective To investigate the expression of calpain small subunit 1 (Capn4) in breast cancer and its clinical significance. Methods The cancer tissues and the paraffin-embeded specimens of the corresponding adjacent normal tissues from 12 patients with breast cancer between May 2015 and July 2015 in Shanghai General Hospital were collected. The expression of Capn4 was detected by using immunohistochemistry. The paraffin-embeded specimens of cancer tissues from 70 breast cancer patients in Shanghai General Hospital between January 2009 and September 2011 were also collected. And the expression of Capn4 was detected. The correlation between Capn4 expression level and TNM stage, pathological grade, molecular markers as well as the prognosis of breast cancer was also analyzed. Results The expression of Capn4 in breast cancer tissues was higher than that in the corresponding adjacent normal tissues. The expression level of Capn4 was correlated with TNM stage (P = 0.002), T stage (P = 0.004), N stage (P = 0.004), M stage (P = 0.025), estrogen receptor (ER) status (χ 2 = 4.787, P = 0.029) and survival status (χ 2 = 5.826, P = 0.016). Kaplan-Meier survival analysis revealed that the median survival time in the higher expression of Capn4 group 90 months was shorter than that in the lower expression of Capn4 group (not reaching the median survival time), and there were statistical differences (χ 2 = 4.351, P = 0.037). Conclusions The expression of Capn4 in breast cancer tissues is upregulated. The expression difference is correlated with TNM stage, pathological grade, ER status and the prognosis of breast cancer, suggesting that it may become a new target and molecular marker for breast cancer treatment.
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The mechanism for icaritin to improve postmenopausal osteoporosis (PMOP) has not been clarified. The aim of this study was to investigate the role of estrogen receptor α36 (ERα36) in the proliferation promotion and anti-apoptosis effects of icaritin on osteoblasts and the underlying mechanism of downstream signal transduction. The ERα36 knockdown human osteosarcoma MG63 cell model was constructed by transfection of shRNA vector. Cell proliferation was detected by CCK-8, the apoptosis was detected by flow cytometry, and the activation of ERK and AKT signaling pathways was detected by Western blot. The results showed that the effects of icaritin on the proliferation and apoptosis of MG63 cells were significantly decreased after ERα36 knockdown, and icaritin could up-regulate the levels of ERK and AKT phosphorylation in MG63 cells, which could be reduced by ERα36 knockdown. The effect of icaritin on the proliferation of MG63 cells was significantly decreased by pretreating the cells with U0126 (an ERK signaling pathway blocker) and LY294002 (an AKT signaling pathway blocker), respectively. Furthermore, anti-apoptotic effect of icaritin on MG63 cells was significantly decreased after the cells were pretreated with U0126, but not with LY294002. These results suggest that icaritin exerts proliferation promotion and anti-apoptosis effects on osteoblasts through ERα36 and its downstream ERK and AKT signaling pathways.
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@#Objective To observe the effects of Pilates on balance, walking and lower limbs motor function in older patients after stroke. Methods From November, 2016 to December, 2017, 60 older patients after stroke were divided into control group (n=30) and Pilates group (n=30). The control group received routine medicine and rehabilitation, the Pilates group accepted Pilates training in addition, for ten weeks. They were evaluated with Berg Balance Scale (BBS), 10-metre Maximum Walking Speed (MWS), Timed 'Up and Go' Test (TUGT) and Fugl-Meyer Assessment-Lower Extremity (FMA-LE) before, and five weeks and ten weeks after training.Results The BBS score, MWS, TUGT and FMA-LE score all improved ten weeks after training in both groups (t>2.122, P<0.05), and improved more in the Pilates group than in the control group (t>2.264, P<0.05).Conclusion Pilates can improve the balance, walking and lower limbs motor function in older patients after stroke.
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Objective To observe the effects of aerobic training and aerobic combined with resistance training on motor function, like muscle strength,cardiopulmonary endurance and so on,in patients with chronic kidney disease(CKD). Methods From July,2015 to August,2016,60 patients with CKD were randomly divided into control group(A,n=20), aerobic training group(B,n=20)and aerobic combined with resistance training group(C,n=20).Group B per-formed cycle ergometer at 50% peak oxygen uptake(VO2peak)for 30 minutes a time,and group C performed one section of Thera-Band resistance training based on group B, three times a week for twelve weeks.All patients were evaluated with one repetition maximum-upper limb (1 RM-U), one repetition maximum-lower limb (1 RM-L),Cardiopulmonary Exercise Test(CPET),Arm Curl Test(ACT),30-second Chair Stand(CS-30),Six-Min-ute Walk Test(6MWT),and estimated gomerular filtration rate(eGFR)and serum creatinine(sCr)were calculat-ed and recorded before and after training. Results There was no significant difference in all indexes among three groups before training(F<1.841,P>0.05).After training,all indexes improved in groups B and C(t>2.162,P<0.05),and were better in groups B and C than in group A(t>2.132, P<0.05).After training, 1 RM-U, 1 RM-L, VO2peak,ACT, CS-30 and 6MWT were better in group C than in group B(t>2.081,P<0.05). Conclusion Aerobic training could improve the motor function of patients with CKD,and it is more effective combined with resistance training.
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Objective To construct the silencing vector of FOS like antigen 1 (FOSL1) gene, and study the effects of FOSL1 on cell proliferation, cell invasiveness and the methylation level of PRDM10 gene in breast cancer cell line MDA-MB-231. Methods The FOSL1 silencing vector of gene pLVX-shRNA-FOSL1-shRNA was purchased. The FOSL1 silencing vector and the empty vector were separately transfected into MDA-MB-231, which were regarded as transfection group and empty group, respectively. Untransfected MDA-MB-231 was used as control group. FOSL1 was verified by PCR in MDA-MB-231. The cell proliferation ability and cell invasion ability of MDA-MB-231 were detected by MTT and Transwell assay, respectively. MSP was used to detect the methylation status of PRDM10 gene. The mRNA and protein expression levels of PRDM10 gene were detected by Q-PCR and Western-blot assay. Results MTT results showed that the optical density (OD) values were significantly lower in transfection group compared with those of control group at 24 h, 48 h and 72 h (all P<0.05), and the same as those compared with empty group at 48 h and 72 h (both P<0.05). Compared with empty group and control group, Transwell assays showed that the cell invasive abilities of MDA-MB-231 were decreased in transfection group (both P<0.05), and MSP assay showed that the methylation of PRDM10 gene was decreased in MDA-MB-231, and Q-PCR and Western-blot tests showed that the expressions of PRDM10 gene were increased in mRNA level and in protein level. Conclusion Silencing of FOSL1 gene inhibits the proliferation and invasion of MDA-MB-231 cells, which might be related to the demethylation of PRDM10 gene in the cells.
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Objective To investigate the effect of the over-expressed CTCF on apoptosis factors Bax and Bcl-2 in human breast cancer cell line MDA-MB-231. Methods Reverse transcription-polymerase chain reaction (RT-PCR) were used to detect the expressions of CTCF,Bax and Bcl-2 in MDA-MB-231. The overexpression vector of CTCF/pEGFP-N1 was constructed. The overexpression plasmid CTCF/pEGFP-N1 and the empty vector plasmid pEGFP-N1 were transfected into breast cancer cell line MDA-MB-231 by lentivirus transfection, and the MDA-MB-231 cells were divided into CTCF group and control group. After successfully transfection of MDA-MB-231 identified by RT-PCR, real time quantitative PCR (Q-PCR) was used to detect the mRNA levels of Bax and Bcl-2 in MDA-MB-231 of the CTCF group and the control group. The protein levels of Bax and Bcl-2 were detected by Western blot assay and enzyme-linked immunosorbent assay (ELISA). Results The expression of CTCF was not found in MDA-MB-231, and expressions of Bax and Bcl-2 were found in MDA-MB-231. Results of Q-PCR showed that the mRNA levels of Bax were 4.63±1.08 and 2.27±0.16 in CTCF group and control group, respectively, and they were statistically significant (t=27.50, P<0.05). The mRNA levels of Bcl-2 were 1.39±0.14 and 3.56 ± 0.97 in CTCF group and control group, and there was significant difference between two groups(t=39.00, P<0.05). Results of Western blot assay showed that the protein level of Bax was higher in CTCF group compared with that of control group. The protein level of Bcl-2 was lower in CTCF group compared with that of control group. Results of ELISA showed that the protein levels of Bax were 15.25±2.17 and 6.24±1.78 in CTCF group and control group, respectively, and there was significant difference between the two groups (t=26.84, P<0.05). The protein levels of Bcl-2 were 4.59 ± 0.97 and 10.68 ± 1.93, and there was significant difference between the two groups (t=21.72, P<0.05). Conclusion The over-expressed CTCF can promote the expression of apoptotic factors and inhibit the expression of anti-apoptotic factors in breast cancer cells.
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Objective To investigate the effect of the over-expressed CTCF on apoptosis factors Bax and Bcl-2 in human breast cancer cell line MDA-MB-231. Methods Reverse transcription-polymerase chain reaction (RT-PCR) were used to detect the expressions of CTCF,Bax and Bcl-2 in MDA-MB-231. The overexpression vector of CTCF/pEGFP-N1 was constructed. The overexpression plasmid CTCF/pEGFP-N1 and the empty vector plasmid pEGFP-N1 were transfected into breast cancer cell line MDA-MB-231 by lentivirus transfection, and the MDA-MB-231 cells were divided into CTCF group and control group. After successfully transfection of MDA-MB-231 identified by RT-PCR, real time quantitative PCR (Q-PCR) was used to detect the mRNA levels of Bax and Bcl-2 in MDA-MB-231 of the CTCF group and the control group. The protein levels of Bax and Bcl-2 were detected by Western blot assay and enzyme-linked immunosorbent assay (ELISA). Results The expression of CTCF was not found in MDA-MB-231, and expressions of Bax and Bcl-2 were found in MDA-MB-231. Results of Q-PCR showed that the mRNA levels of Bax were 4.63±1.08 and 2.27±0.16 in CTCF group and control group, respectively, and they were statistically significant (t=27.50, P<0.05). The mRNA levels of Bcl-2 were 1.39±0.14 and 3.56 ± 0.97 in CTCF group and control group, and there was significant difference between two groups(t=39.00, P<0.05). Results of Western blot assay showed that the protein level of Bax was higher in CTCF group compared with that of control group. The protein level of Bcl-2 was lower in CTCF group compared with that of control group. Results of ELISA showed that the protein levels of Bax were 15.25±2.17 and 6.24±1.78 in CTCF group and control group, respectively, and there was significant difference between the two groups (t=26.84, P<0.05). The protein levels of Bcl-2 were 4.59 ± 0.97 and 10.68 ± 1.93, and there was significant difference between the two groups (t=21.72, P<0.05). Conclusion The over-expressed CTCF can promote the expression of apoptotic factors and inhibit the expression of anti-apoptotic factors in breast cancer cells.
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Objective Serum uric acid (SUA) levels have been identified to be associated with nonalcoholic fatty liver disease (NAFLD) which is often closely linked to obesity. However, few studies have examined the relationship between SUA levels and NAFLD in normal body mass index (BMI) people in whom NAFLD is noted not uncommon. We conducted this study to assess the association of SUA with NAFLD in Chinese normal-BMI people. Methods A total of 2 193 normal-BMI people (1 106 men and 1087 women) who participated in a health check-up were enrolled in this cross-sectional study. NAFLD was defined as a hepatic steatosis examined by liver ultrasonography in the absence of a second cause. Multiple regression analysis was performed to examine the relationship between SUA and NAFLD. Results Compared with people without NAFLD, those with NAFLD had significantly higher level of serum uric acid [males,(339.0±47.6)μmol/L vs.(309.3±53.5)μmol/L, t=3.296, P<0.01;female,(273.6±41.6)μmol/L vs. (243.9 ± 47.6)μmol/L, t=3.178, P<0.01]. Multiple regression analysis revealed that the quartile 3 and 4 of SUA in both sexes had a higher (P<0.05) odds ratio for the presence of NAFLD than the lowest quartile [males,odds ratio:2.508(1.255-3.374)and 3.383(2.058-5.490);females,odds ratio:2.186(1.090-4.384) and 2.420(1.214-4.821),respectively],after adjusting for age, body mass index, blood urea nitrogen, serum creatinine, the liver-function test, metabolic components, high sensitivity C-reactive protein, and smoking status.Conclusions Increased SUA level, even within the normal range, are independently associated with the presence of NAFLD among normal-BMI people.
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@# Community rehabilitation of cardiovascular disease is of great significance. Exercise therapy of cardiovascular disease in communities is necessary and feasible. At present, the main problems China faced to carry out community exercise therapy of cardiovascular disease include: cardiovascular patients with a weak sense of community exercise therapy, lack of professional rehabilitation personnel, inadequate community-based exercise therapy practice and lack of community rehabilitation funding. To solve above problems, we should strengthen the community exercise therapy practice of cardiovascular disease, and promote the cooperation between community rehabilitation and hospital rehabilitation. In all, comprehensive efforts should be made in order to solve the problems.
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<p><b>OBJECTIVE</b>The phosphatidylinositol-3-kinase (PI3K)-AKT signaling pathway is considered to play an important role in tumorigenesis. Frequent somatic mutations in the PI3K subunit p110a (PIK3CA) occur in a variety of cancer types. The purpose of this study was to determine the relationship between PIK3CA mutation in breast cancer and pathological features and outcome of patients.</p><p><b>METHODS</b>The PIK3CA mutations in exons 7, 9, 20 were screened in 250 primary breast cancers using PCR and fluorescent (F)-SSCP, and the results were analyzed according to their cliniopathological data.</p><p><b>RESULTS</b>The frequency of PIK3CA mutations among the 250 cases was 35.2% (88/250), point mutations in exon 7 were found in 8 (3.2%) cases,40 (16.0%) cases in exon 9 and 47 (18.8%) cases in exon 20. No significant correlation between PIK3CA mutation and age, histological type, differentiation, and lymph node metastasis was observed. Mutations were associated with larger tumor size (P = 0.004) and positive estrogen receptor status (P = 0.008). Patients with PIK3CA mutations showed a significantly worse survival (P = 0.004), particularly in those with positive estrogen receptor expression or non-amplified HER-2 (both P = 0.002).</p><p><b>CONCLUSIONS</b>PIK3CA mutations may play an important role in the carcinogenesis and development of breast cancer. The association with large tumor size, ER+ and poor survival indicates that PIK3CA mutation could be an independent factor for tumor malignant phenotype and prognosis.</p>
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Adolescent , Adult , Aged , Aged, 80 and over , Female , Humans , Middle Aged , Young Adult , Adenocarcinoma , Genetics , Metabolism , Pathology , Breast Neoplasms , Genetics , Metabolism , Pathology , Carcinoma, Ductal, Breast , Genetics , Metabolism , Pathology , Carcinoma, Lobular , Genetics , Metabolism , Pathology , Carcinoma, Medullary , Genetics , Metabolism , Pathology , Class I Phosphatidylinositol 3-Kinases , Exons , Follow-Up Studies , Lymphatic Metastasis , Phosphatidylinositol 3-Kinases , Genetics , Metabolism , Point Mutation , Receptor, ErbB-2 , Metabolism , Receptors, Estrogen , Metabolism , Survival Rate , Tumor BurdenABSTRACT
Objective To study the effectiveness and feasibility of low intensity of aerobic bicycle ergometer training on hemiplegic stroke patients unable to perform assistive or independent ambulation.Methods Thirty stroke patients unable of assistive or independent ambulation were randomly divided into a control group and a aerobic training group.Patients in the aerobic training group performed low intensity of aerobic bicycle ergometer training for 30 min,3 times a week for 6 weeks in addition to a routine rehabilitation regime.The training duration and course of routine rehabilitation training were the same in both groups.The related cardiovascular response in aerobic training and changes in motor performance as well as the risk factors for stroke were observed in both groups.Results Twenty-four patients(12 patients in each group)completed the experiment.Patients in aerobic training group performed all training regime safely without any adverse response.Their motor function(including FMA score,Rivermead index,BI ADL ability),exercise endurance duration in exercise test,glucose level at 2 h in oral glucose tolerance test,fasting insulin level and HOMA-IR index all improved greatly as compared to the control group(P ＜ 0.05).Conclusion Low intensity of aerobic bicycle ergometer training can be safely and effectively administered in hemiplegic stroke patients unable of assistive ambulation or independent ambulation.
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Rehabilitation therapy with the integrative traditional and western medicine is the devel opment trend in China,and it is also the aim of talent education of Nanjing University of Chinese Medicine.Based on the introduction of rehabilitation therapy education in our university,this article makes a discussion on the research and practice of curriculum reformation of rehabilitation therapy with the integrative traditional and western medicine.Additionally,it puts forward some implement measures.
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The purpose of this investigation is to improve ethanol production and decrease acetate formation in Saccharomyces cerevisiae strain YS2-?adh2.The strain YS2-?adh2 with deleted alcohol dehydrogenase Ⅱ(adh2) gene was isolated in our lab with higher ethanol production than that of the strain YS2.The ace-taldehyde dehydrogenase Ⅵ(ald6) gene encoded a cytosolic acetaldehyde dehydrogenase,a key enzyme of the pyruvate dehydrogenase(PDH) bypass,transfers acetaldehyde to acetate.To disrupt ald6 gene of the strain YS2-?adh2,ald6 gene targeting cassettes were synthesized by long flanking homology PCR(LFH-PCR) and then were transformed into YS2-?adh2 mutants by LiAc/SS Carrier DNA/PEG method.Positive transformants were selected with G418 and further confirmed by PCR.Once correctly integrated into the genome,the selective marker was rescued by transforming the plasmid pSH65 into the positive transformants and inducing the Cre expression with a Cre/loxP-mediated marker removal procedure.We named the ald6 gene knocked-out strain as YS2-?adh2-?ald6 which has a 12.5% higher ethanol production and a 18% lower acetate formation compared to the strain YS2.
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The sfa1 gene encoded a bifunctional enzyme with the activities of both alcohol dehydrogenase and glutathione-dependent formaldehyde dehydrogenase in Saccharomyces cerevisiae.The gene disruption cassette produced by PCR using the same long oligonucleotides which comprise 19 or 22 nucleotides complementary to sequences in the templates(pUG6 and pUG66 marker plasmid)at 3' end and 45 nucleotides at 5' end that annealed to sites upstream or downstream of the genomic target sequence to be deleted.After two linear disruption cassettes with a Cre/loxP mediated marker were transformed into the cells of Saccharomyces cerevisiae YS-1,the positive transformants were checked by PCR to correct the integration of the cassette and concurrent deletion of the chromosomal target sequence.Once correctly integrated into the genome,the select marker can be efficiently rescued by transformating the plasmid pSH47 into YS-1 and inducing the Cre expression with a Cre/loxP-mediated marker removal procedure.The expression of the Cre recombinase finally resulted in the removal of the marker gene,leaving behind a single loxP site at the chromosomal locus.The diploid mutant YS-1-sfa1 was generated,which could enhance the output of ethanol with 8.0% by shaking culture in flask compared with the original strain YS-1.