AIM:To clarify the impact of heart shock factor 4b (Hsf4b) K65R mutation on the regulation of downstream protein expression .METHODS:Non-functional Lys mutant plasmid pWZL-blast-HA-Hsf4b/K65R was genera-ted by replacing single , homologous amino acids using KOD-Plus-Mutagenesis-Kit.Mouse lens epithelial mLEC stable cell lines expressing Hsf4b or Hsf4b/K65R were constructed by lentivirus infection .The expression of Hsf4b in the mutant and the wildtype mLEC cells was confirmed by Western blotting .The expression of Hsf4b downstream proteins such as heat shock protein ( Hsp)70, Hsp90, Hsp27 and CryAB was examined by Western blotting and real-time PCR.RESULTS:The results of PCR and DNA sequencing confirmed the successful construction of mLEC Hsf 4b/K65R mutant.The K65R mutation didn’t influence Hsf4b expression in the mLEC cells.After K65R mutation in Hsf4b, the expression levels of Hsp27 and CryAB were down-regulated and the expression of Hsp 70i and Hsp90a upregulated.CONCLUSION: pWZL-blast-HA-Hsf4b/K65R can be used to construct a stable cell line by infecting with lentivirus .Hsf4b/K65R mutation influ-ences the regulation of downstream heat shock proteins .

Objective To investigate the effects of dopaminergic medication on decision-making un-der ambiguity in patients with early Parkinson's disease( PD) . Methods Using Iowa Gambling Task ( IGT) for 24 early non-medication idiopathic PD patients( Hoehn and Yahr Scale≤Ⅱlevel) ,24 early idiopathic PD patients with regular dopaminergic medication and also for 24 healthy controls( HC) whose age,gender,and education match to PD patients to test their ability of decision-making under ambiguity. Results The results showed non-medication PD group showed impairments on digtal span and verbal fluency and decision-making task. There was significant difference in IGT task scores among the three groups(F=6.024, P=0.004) . The total net scores of advantageous choices in IGT were significantly lower in non-medication PD group( (-4.50 ±22.19) scores) than medication PD group((8.83±23.24)scores) and healthy group((15.92±15.77) scores) . The difference of net scores in block1 to block5 between non-medication PD group and medication PD group was gradually increased,and the difference of net scores in block5 was significant(P<0.05) . There was no significant difference in total net scores and net scores in block1 to block5 between medication PD group and healthy group(P>0.05) . As the game processing,medication PD group gradually shifted their se-lections toward the advantageous choices. But non-medication group did not exhibit this shift pattern and the performance was much poorer. Meanwhile, the study also indicated the total net scores of advantageous choices for non-medication PD group was positive correlation to the MoCA scores ( r=0.614, P=0.001). Conclusion The present study has shown that non-medication PD group has impairment in decision-making under ambiguity risk condition and prefer to choose risky options. when exogenous complement dopaminergic medication,the risk decision-making ability of medication PD group has been improved.

Objective To compare the effects of different extracts of Rheum palmatum on weight and tissue structure of hypothalamusand and pituitary of adult female rats,and screen the main reproductive toxicity extract.Methods The water,chloroform,ethyl acetate,and n-butanol extracts and water-soluble substance of R.palmatum were prepared by polarity gradient extraction method.Female adult rats were randomly divided into blank control group,rhubarb water extract group,and different extracts groups.The dosage of all the groups was equivalent to 4.00 g/kg crude rhubarb.Rats were administered with extracts by gavage for 60 d.Body mass growth rate of rats were calculated before and after administration.The pathological changes of hypothalamic arcuate nucleus neurons and pituitary gonadal cell were observed with light microscope.Results Compared with the blank control group,the body mass growth rate of rhubarb water extract group was decreased (P ＜ 0.05),while those in the different extracts groups were increased (P ＜ 0.01);The hypothalamic arcuate nucleus neurons of rhubarb water extract group showed chromatin marginalization,nissl substance dissolving,fuzzy boundary of nuclear membrane,as well as hell cells,and the total number of adenohypophysis cells reduced and the cells arranged in irregular.However,there were no apparent pathologic changes in different extracts groups.Conclusion Rhubarb water extract administration by long-term dose can reduce weight growth rate and result in pathologic changes of hypothalamic arcuate nucleus and adenohypophysis,while the different extracts can increase weight growth rate significantly and have little effects on the organizational structure ofhypothalamic arcuate nucleus and adenohypophysis.

To explore the serum miR-338-5p expression characteristics in renal transplant recipients ,and the role of regulating BAFF signal ,then investigate its biological significance.Methods:Healthy volunteers were enrolled as control group.Serum miR-338-5p was detected by real-time PCR;soluble BAFF was detected by ELISA;anti-HLA-Ⅰantibody,anti-HLA-Ⅱ antibody and anti-MICA antibody were detected by liquid chip technology.SPSS17.0 software was applied.t-test was used to compare the means of two independent samples;Paired samples t-test was used to compare the means of two paired samples;Spearman method and Pearson method were used to analyse the correlation;P<0.05 was considered to be statistically significant.Results: Compared with healthy controls,serum miR-338-5p in renal transplant recipients decreased significantly (P<0.001),while serum BAFF increased significantly (P<0.01).Serum miR-338-5p levels within 1 year post-transplantation were significantly lower than that of more than 1 year post-transplantation (P<0.01);Serum miR-338-5p levels within 3 years post-transplantation were significantly lower than that of more than 3 years post-transplantation (P<0.01);To all research objects,serum miR-338-5p was significantly negatively correlated with serum BAFF (r=-0.51,P<0.001),and serum miR-338-5p was significantly negatively correlated with anti-HLA-Ⅱ antibody(r=-0.322, P<0.05);Serum miR-338-5p within 3 years was significantly negatively correlated with anti-HLA-Ⅱantibody (r=-0.423,P<0.05), and serum miR-338-5p within 3 years was significantly negatively correlated with anti-MICA antibody(r=-0.411,P<0.05);Serum miR-338-5p more than 3 years was significantly positively correlated with anti-MICA antibody(r=0.486,P<0.05),and Serum miR-338-5p more than 3 years was significantly positively correlated with anti-HLA & MICA antibody(r=0.578,P<0.01).Conclusion:miR-338-5p may directly or indirectly target BAFF signal ,and participate antibody mediated immune response by regulating its target genes and interfere with the long-term survival of transplanted renal.

Objective:To investigate the effect of the application of "internet +" disease management based on the AISAS model on the young and middle-aged patients after PCI therapy.Method:A total of 90 young and middle-aged patients hospitalized in cardiological department of the First Affiliated Hospital of University of Science and Technology of China (Anhui Provincial Hospital) were enrolled between June 2018 to October 2019, and they were randomly divided into observation group and control group with 45 cases in each group. Patients in the control group received telephone follow-up while "internet +" disease management based on the AISAS model was applied in the observation group. The drug therapy compliance, acquisition of the disease knowledge, quality of life, anxiety and depression levels, return to workand incidence of cardiovascular events.Results:One year after the intervention, the drug therapy compliance score, disease-related knowledge score, quality of life score, SAS and SDS scores of the observation group were 7.55±1.21, 7.29±1.27, 701.17±74.86, 32.55± 4.31, 34.74±4.16, the scores of patients in the control group were 6.48±1.56, 6.12±1.94, 670.58±65.29, 41.72±4.33, 40.79±4.17. The difference was statistically significant ( t value was 2.066-11.203, P<0.05). The comparison between the return of the patients and the incidence of cardiovascular events in the two groups was statistically significant ( χ2 value was 5.031, 11.275, P<0.05). Conclusion:This management model can increase the knowledge of disease PCI postoperative patients, improve their quality of life, make patients return to society earlier, and promote the improvement of the quality of continuous nursing service.

OBJECTIVE:To determine the contents of citric acid in fentanyl citrate raw materials and its injection by ion chro-matography. METHODS:The determination was performed on Thermo Dionex IonPacTM AS11-HC column with mobile phase con-sisted of potassium hydroxide (gradient elution) at the flow rate of 1.0 mL/min. The column temperature was 35 ℃,and sample size was 20 μL. The detector was suppressed conductivity detector. RESULTS:The linear range of citric acid were 0.1157-74.05 μg/mL(r=0.9995). The limit of quantitation was 0.1150 μg/mL,and the limit of detection was 0.0400 μg/mL;RSDs of preci-sion,stability and reproducibility tests were all lower than 2.0%;the average recoveries were 99.6%-101.5%(RSD=0.68%,n=9). CONCLUSIONS:The method is environmentally-friendly and simple with good accuracy and precision,and suitable for the contents determination of citric acid in fentanyl citrate raw materials and injection.

Objective To investigate the prevalence of GJB2 ,SLC26A4 and mitochondrial DNA 12S rRNA m .1555A>G(mtDNA 1555A>G) mutations in Hui ethic group patients with nonsyndromic hearing loss (NSHL) from Northwest China .Methods A total of 420 peripheral blood samples were collected from unrelated Hui ethic group probands with NSHL in Northwest China .Amplified the target gene by polymerase chain reaction (PCR) af-ter extracting genomic DNA from whole blood .The mtDNA 1555A>G mutation was detected by PCR -Alw26I di-gestion ,then direct sequencing was used to the positive samples of mtDNA 1555A> G ,the coding region of GJB2 gene ,exon 8 and 19 of SLC26A4 gene .Results There were 11(2 .62% ) cases caused by mtDNA 1555A>G homo-zygous mutation in 420 patients with NSHL .There were 41(9 .76% ) cases including homozygote and compound het-erozygote ,caused by GJB2 gene mutation ,which was the most frequent deafness -related gene .The allel frequency of c .235delC accounted for 6 .90% ,as well as the most frequent(51 .33% ) mutational pattern in GJB2 gene .There were 20 patients(4 .76% ) were found carring two allel mutations in SLC26A4 gene .The allel frequency of c .919 -2A>G was 5 .0% ,accounting for a total of 68 .85% in all base alterations of SLC26A4 gene ,which was the major mutant form of SLC26A4 gene .Conclusion GJB2 gene is the most common deafness -gene in Hui ethnic group pa-tients with NSHL from Northwest China ,while c .235delC is the main mutant form ,and c .919-2A>G is the hot-spot mutation of SLC26A4 gene .Through this study we can provide the molecular epidemiology basis for Hui ethnic group patients with NSHL from Northwest China in genetic diagnosis ,genetic counseling and therapy by associated testing of three frequent hearing loss genes .

[ ABSTRACT] AIM: To investigate the protective effect of autophagy on oxidized low density lipoprotein ( ox-LDL)-induced macrophage apoptosis and the underlying molecular mechanisms .METHODS:The RAW264.7 macropha-ges were pretreated with 3 mmol/L 3-methyladenine (3-MA), 1 μmol/L rapamycin (Rap) or 4 mmol/L 4-phenylbutyric acid ( PBA) respectively for 1 h and then treated with ox-LDL (100 mg/L) for 12 h.The cell viability and apoptosis were determined by MTT assay and flow cytometry with Annexin V-FITC/PI staining, respectively.The activities of lactate de-hydrogenase ( LDH) in the medium and caspase-3 in the cells were determined by detection kits .The protein levels of bec-lin-1 (a molecular marker of autophagy ), glucose-regulated protein 78 (GRP78, an endoplasmic reticulum stress marker) and C/EBP homologous protein ( CHOP, a key-signaling component of endoplasmic reticulum stress-induced apoptosis ) were examined by Western blot .Microtubule-associated protein 1 light chain 3 (LC3, another molecular marker of autoph-agy) was observed under laser scanning confocal microscope .RESULTS: Treatment of the RAW264.7 macrophages with ox-LDL at 100 mg/L for 12 h resulted in significant decrease in cell viability , and dramatic elevation in LDH leakage , cell apoptosis and caspase-3 activity, which were promoted by 3-MA (an autophagy inhibitor) and inhibited by Rap (an autoph-agy inducer ) .ox-LDL induced autophagy in the macrophages as assessed by beclin-1 upregulation and frequent granulation of LC3, which were inhibited by 3-MA and promoted by Rap.Interestingly, 3-MA enhanced, while Rap blocked, the CHOP upregulation induced by ox-LDL.Moreover , PBA ( endoplasmic reticulum stress inhibitor ) significantly inhibited ox-LDL-induced GRP78 upregulation and autophagy as determined by the attenuation of beclin-1 upregulation and frequent granula-tion of LC3.CONCLUSION: Endoplasmic reticulum stress mediates ox-LDL-induced autophagy in macrophages , and moderates activation of autophagy may protect macrophages from ox-LDL-induced apoptosis by inhibiting CHOP expression .

The standardized training is an indispensible stage for the improvement of residents' comprehensive quality and for the training of high-qualified talents.The article preliminarily explored the standardized training model for residents,which was in accordance with the characteristics of the department of cardiology mainly from four aspects:the set-up of reasonable training program,the training of practical skills,the training of humanistic quality and the training of life-long learning ability.

Objective: To investigate the characteristics of histogram analysis and differences of apparent diffusion coefficient (ADC) for normal pancreas with different diffusion gradients of diffusion weighted imaging (DWI) at 3.0T. Methods: Three unidirectional diffusion gradients (X, Y or Z direction) and three orthogonal diffusion gradient DWI data of 21 volunteers with normal pancreas were retrospectively analyzed. The histogram parameters of mean ADC, percentile, skewness, peak and the like were measured based on home-made DWI post-processing software, and the differences on histogram parameters obtained from 4 different diffusion gradients were compared by using Kruskal Wallis test. Results: There was no significant difference on the mean ADC, skewness, percentile and the like for normal pancreas among 3 unidirectional diffusion gradients and tri-orthogonal diffusion gradient DWIs, but there was significant difference on the mean kurtosis value (X: 3.16±1.44, Y: 4.72±2.26, Z: 4.47±2.1, tri-orthogonal: 3.69±1.82; χ² =-9.84, P=0.02). Further analyses results showed that the ADC kurtosis of normal pancreas obtained from X-directional diffusion gradient DWI was significantly smaller than that of the direction of Y or Z, and the difference was statistically significant (P<0.05). Conclusions The ADC kurtosis of normal pancreas is related to the direction of diffusion gradient in DWI, and the ADC kurtosis of normal pancreas obtained by X-direction diffusion gradient is the smallest.