The aim of this study is to construct specific shRNA expressing plasmids, and to observe their effects on H9c2 cardiomyocytes injury induced by hypoxia/reoxygenation (H/R). RIPK1 and RIPK3 are the key kinases mediating the process of necroptosis. Using recombinant DNA technology, we inserted the synthetic shRNA into pSUPER vector to construct RIPK1-shRNA or RIPK3-shRNA plasmid respectively. We transfected H9c2 cardiomyocytes with the two shRNA plasmids respectively, before we treated them with H/R stimulation. Then, we measured the relevant genes and proteins by real-time PCR and Western blot. Meanwhile,we detected the markers of necroptosis and cardiomyocytes injury. The results showed that inhibition of ripk1 or ripk3 gene expression by its specific shRNA might protect the cardiomyocytes injury induced by H/R stimulation.
Animals ; Apoptosis ; Cell Hypoxia ; Cell Line ; Gene Expression ; Myocytes, Cardiac ; pathology ; Protein-Serine-Threonine Kinases ; genetics ; metabolism ; RNA, Small Interfering ; Rats ; Real-Time Polymerase Chain Reaction ; Receptor-Interacting Protein Serine-Threonine Kinases ; genetics ; metabolism ; Transfection

The aim of this study was to observe whether necroptosis is involved in the process of cardiac hypertrophy induced by pressure overload. SD rats underwent transverse abdominal aortic constriction (TAC) operation for establishing cardiac hypertrophy model. The structure and function of the left ventricle of rats were evaluated via echocardiography, left ventricular mass index, the expression of markers of cardiac hypertrophy and histological detection. Real-time PCR and Western blot were used to measure the gene and protein expression of receptor interacting protein kinase 1 and 3 (RIPK1 and RIPK3, the necroptosis markers) respectively. Four weeks after TAC operation, rat model for cardiac hypertrophy was established. The experimental data showed that the gene and protein expressions of RIPK1 and RIPK3 in the rat heart hypertrophic tissues after TAC for 4 weeks were increased significantly compared with those in the sham group. HE staining showed cardiomyocytes injury and hypertrophy in the hearts of TAC rat models. By transmission electron microscope, we observed that mitochondria of cardiomyocytes were damaged seriously in the TAC models. Treatment with losartan used, the selective antagonist of angiotensin II type I receptor could improve the cardiac function of TAC rats. Moreover, losartan treatment decreased the expression of RIPK1 and RIPK3 in heart tissues of TAC rats. The results suggest that necroptosis occurrs in the process of cardiac hypertrophy with pressure overload, and losartan could alleviate the cardiac hypertrophy and inhibit necroptosis.
Animals ; Apoptosis ; Cardiomegaly ; pathology ; Disease Models, Animal ; Echocardiography ; Heart ; physiopathology ; Losartan ; pharmacology ; Myocytes, Cardiac ; Pressure ; Protein-Serine-Threonine Kinases ; metabolism ; Rats ; Rats, Sprague-Dawley ; Real-Time Polymerase Chain Reaction ; Receptor-Interacting Protein Serine-Threonine Kinases ; metabolism

Objective To analyze the influence of hepatosteatosis on pancreatic β-cell function in type 2 diabetes mellitus (T2DM).Methods A total of 213 subjects with T2DM from Metabolic Disease Hospital,Tianjin Medical University from January 2013 to December 2013 were included in the study.Non-alcoholic fatty liver disease (NAFLD) was diagnosed with abdominal ultrasonography.Patients were divided into two groups:T2DM with NAFLD and T2DM without NAFLD.ALT,AST,γ-glutamyltransferase,serum lipid,glycosylated hemoglobin A1 c (HbA1c),fructosamine,fasting glucose,insulin and 2 hours plasma glucose,insulin after 75g glucoseload were detected.The insulin resistance and β-cell function were assessed by HOMA-IR and HOMA-β.Results Among the 213 T2DM subjects,51％ (108 cases) were with NAFLD.The HOMA-IR [4.76 (2.83,7.21) vs 2.79 (1.76,4.37),P ＜ 0.05] and HOMA-β [49.18 (37.78,85.09) vs 29.50 (18.09,45.54),P ＜ 0.05] were significantly higher in T2DM with NAFLD than those in T2DM alone.Within subjects with T2DM and NAFLD,the HOMA-IR [6.28 (2.87,8.17) vs 2.95 (2.07,3.66) P ＜ 0.05] and HOMA-β [59.18 (37.78,85.09) vs 30.59 (28.56,34.49),P ＜ 0.05] levels were higher in subjects with normal liver function than those with abnormal liver function.Conclusions T2DM patients with NAFLD have severer insulin resistance than those without NAFLD.The β-cell function of those patients was compensatory increased,which was decreased in subjects with abnormal liver function.

Objective:To investigate the expression of LAIR-1 on Treg cells and TGF-βin the peripheral blood of patients with oral lichen planus(OLP).Methods:The peripheral blood was obtained from 22 OLP patients and 22 healthy controls.Flow cytometry was used to evaluate the proportion of LAIR-1 on CD4 +Foxp3 +Treg cells,and the level of TGF-βwas measured by ELISA.Results:In the peripheral blood of OLP patients,the proportion of LAIR-1 on CD4 +Foxp3 +Treg cells and TGF-βcontent were significantly higher than those of the healthy controls(P ＜0.05),The proportion of LAIR-1 on CD4 +Foxp3 +Treg cells was positively correlated with the level of TGF-β(r =0.43,P ＜0.05).Conclusion:The high expression of LAIR-1 on CD4 +Foxp3 +Treg cells and TGF-βin the pe-ripheral blood of OLP patients may be responsible for the pathogenesis of OLP.

OBJECTIVE: To explore the found ways and first diagnosis age of children with large vestibular aqueduct, and their relations with hearing loss. METHOD: Medical histories of 122 cases of children diagnosed with large vestibular aqueduct by HRCT or MRI had been collected from January 2009 to April 2014 in our hospital children's hearing diagnosis center clinic. Found ways comprise of accepting universal newborn hearing screening (UNHS) group and unaccepting UNHS group. Accepting UNHS children were divided into two ears unpassing group, single ear unpassing group and passing group. The patients in unaccepting UNHS group were divided into not sensitive to sounds, speech stunting, sudden hearing loss, and other group. Analysis the relationship between the found ways and first diagnosis age and their relations with hearing loss. RESULT: There are 84 cases (68.85%) accepting UNHS, the average age of first diagnosis was (17.24 ± 17.08) months; 37 cases (31.15%) are not accepting UNHS. The average age of first diagnosis was (30.92 ± 18.21) months. The average first diagnosis age of accepting UNHS group was more earlier than the unaccepting UNHS group. The difference was statistically signif- icant (P < 0.01). There were 57 cases (67.85%) whose two ears not pass UNHS; 15 cases (17.86%) single ear not pass; namely the referral rate was 85.71%; 12 cases (14.29%) pass the test. The first diagnosis age of passing UNHS group was more later than two ears unpassing group (P < 0.001). In the unaccepting UNHS group, the average first diagnosis age of not sensitive to sounds group (19.69 ± 11.16 months) was more earlier than words dysplasia group (37.13 ± 15.62 months) and sudden hearing loss group (47.40 ± 24.70 months) (P < 0.01). The difference in the degree of hearing loss between accepting UNHS and unaccepting UNHS group had no statistical significance (P > 0.05). In unaccepting UNHS group ,the average first diagnosis age of the mild-to-moderate hearing loss group was later than the very severe hearing loss group (P < 0.01). CONCLUSION Most of large vestibular aqueduct children can be found and receive diagnosis early by UNHS. But part of these patients with late-onset or progressive hearing loss, especially these with mild-to-moderate hearing loss cannot be found early, which should arouse our attention.
Child, Preschool ; Deafness ; Early Diagnosis ; Hearing Loss ; etiology ; Hearing Loss, Sudden ; Hearing Tests ; Humans ; Infant ; Infant, Newborn ; Neonatal Screening ; Vestibular Aqueduct ; abnormalities

Objective To compare the predicting values for Prognosis among Global Risk Classification (GRS),Synergy Between Percutaneous Coronary Intervention With TAXUS and Cardiac Surgery (SYNTAX) score,the European System for Cardiac Operative Risk Evaluation (EuroSCORE) in patients who received stenting because of unprotected left main coronary artery (ULMCA) lesion.Methods Totally 105 successive elderly patients with ULMCA lesion who received stenting were divided into 2 groups:with and without main adverse cardiac events (MACE).The clinical and angiographic characteristics were analyzed and then compared among GRC,SYNTAX score and EuroSCORE.Results As compared with none MACE group,MACE group had higher EuroSCORE score (2.0±2.3 vs.6.5±2.9,t=8.18,P=0.002),and more trivessel disease and left main bifurcation lesion (x2 =8.96,6.96,P =0.011,P =0.008).High risk GRC showed more MACE than medium or low risk GRC [55.9％ (19/34) vs.20.5％(9/44),7.4％ (2/27),x2 =19.77,P=0.001].AUC(95％CI )of GRC,SYNTAX score and EuroSCORE were [0.821 (0.730-0.912),0.586(0.462-0.709) and 0.631 (0506-0.757)],respectively.Compared with SYNTAX score and EuroSCORE,GRC was superior in the MACE predicting value (Z=3.29,2.63,P＜0.01 or P＜0.05).

Objective To evaluate the value of 3.0 T dynamic contrast-enhanced magnetic resonance imaging(MRI) in the diagnosis of hip joint lesions in patients with ankylosing spondylitis (AS).Methods Fifty-two AS patients with hip involvement were diagnosed in Department of Rheumatology in our hospital from January 2013 to December 2016.All patients underwent AS disease activity ankylosing spondylitis disease activity score (ASDAS)-C-reactive protein (CRP) score evaluation of hip joint acti-vity and hip joint disease activity was evaluated by doctors.According to ASDAS-CRP score,patients were divided into active disease 34 cases and remission for 18 cases.All patients underwent double hip MRI plain scan,dynamic enhanced scan and post-processing analysis after the diagnosis.Six regions of interest were selected in the abnormal synovial area around the hip joint.The time signal TIC curve was generated automatically,the shape of the curve was described and the early intensification rate of the third,fourth phase was measured.The early intensification rates of the active disease group and the remission group were analyzed statistically.Two independent samples t test was used for analysis,and P＜0.05 was regarded as statistically significant.The early intensification rate of the third,fourth phase were compared with the ASDAS-CRP score,the evaluation of hip joint activity in patients and the evaluation of hip joint activity were carried out by doctors taking care of the patients.Results The TIC curve of the active group was rising fast,followed by platform or fast rising.The curve of the remission group was slow rising.The inten-sification rate of the third,fourth phase [(116±9)％,(146±12)％ respectively] in the active phase group was higher than that of the remission group [(23±5)％,(43±14)％],and the difference was statistically significant (t=2.447,3.054;P＜0.05).The rate of synovial enhancement of the third,fourth phases was positively correlated with the ASDAS-CRP score (r=0.802,0.836,P=0.01),and the correlation was poor with evaluation of hip joint activity by patients (r=0.352,0.458;the P=0.02,0.01respectively),and the correlation was goodwith the evaluation of hip joint activity by doctors (r=0.652,0.717;P=0.01).Conclusion The shape and early enhancement rate of magnetic reson-ance dynamic enhanced TIC curve is valuable in assessing the disease activity of hip joint lesions in AS patients.

Objective:To investigate the normal range of fetal ventricles and posterior cranial fossa development in the second and third trimesters and their variations with gestational age using quantitative MRI analysis.Methods:This retrospective study enrolled 675 pregnant women who underwent prenatal MRI examination with an average gestational week of 29.0±8.5 in the Third Affiliated Hospital of Guangzhou Medical University from January 2016 to January 2020. MRI data of all the subjects were collected and analyzed, including left lateral ventricle trigonometric width (LLVTW) and right lateral ventricle trigonometric width (RLVTW), third ventricle width (TVW), fourth ventricle width (FVW), the anterior-posterior diameter of the fourth ventricle (APDFV), cavum septum pellucidum width (CSPW), cisterna magna width (CMW), etc. Spearman, Pearson correlation analysis, and t-test were used for the statistical analysis. Results:(1) Totally 675 fetuses were recruited, including 392 female and 283 male fetuses. No statistical difference of gestational weeks at MRI was found between male and female fetuses. (2)The mean value of TVW and CMW of the female fetuses were significantly higher than those of male fetuses [(0.60±0.05) vs (0.63±0.04) cm, t=－5.059; (0.57±0.14) vs (0.67±0.15) cm, t=－7.445; both P<0.001]. Spearman correlation analysis showed that TVW and CMW were negatively correlated with fetal gender ( r=－0.179 and－0.312, both P<0.001). (3)Pearson correlation analysis showed that LLVTW, RLVTW, TVW, FVW, APDFV, CSPW, and CMW were all positively correlated with gestational weeks ( r=0.310, 0.267, 0.205, 0.801, 0.829, 0.216 and 0.284, all P<0.001). FVW and APDFV were significantly linearly correlated with gestational weeks (r=0.801 and 0.829, both P<0.001). (4) There was no significant change in LLVTW and RLVTW in the second trimester, but a slight increase was found in the third trimester. TVW showed a scattered distribution in the second and third trimesters and increased slightly with the gestational week at 26 to 27 +6 gestational weeks. FVW and APDFV increased linearly while. CSPW increased slowly with gestational weeks in the second and third trimesters. CMW increased slightly with gestational weeks in the second and third trimesters but showed little change in the third trimester. Conclusions:The development of fetal ventricles and posterior cranial fossa in the second and third trimesters show a growth trend of varying degrees with the increase of gestational weeks. TVW and CMW are significantly negatively correlated with the fetal gender.

Objective To investigate the relationship between single b-value and multiple b-value diffusion weighted imaging (DWI)and different types of breast lesions.Methods The data of 92 cases including 8 cases of cyst,11 cases of granulomatous inflammation,33 cases of benign nodules lesions and 40 cases of malignant tumor were analyzed retrospectively,who underwent single b-value DWI and multiple b-value DWI and all patients were confirmed by pathology.The normal side of the mammary gland fiber was taken as the control group.Apparent diffusion coefficient(ADC)were obtained by the monoexponential model.Slow apparent diffusion coefficient(ADCslow),fast apparent diffusion coefficient(ADCfast),the fraction of fast ADC(ffast)were obtained by the biexponential model.ADC,ADCslow,ADCfast,ffastvalues of different breast lesions were analyzed by single factor analysis of variance.The LSD-t test was used to compare the mean values of the parameters of the breast lesions in pairs.Results The ADC and ADCslowvalue of breast lesions were decreased in the order of cyst (2.369 ± 0.380)×10-3mm2/s,(1.294 ± 0.439)×10-3mm2/s,normal gland(1.785 ± 0.298)×10-3mm2/s,(0.946 ± 0.321)×10-3mm2/s,benign tumor(1.647 ± 0.296)×10-3mm2/s,(0.944 ± 0.256)×10-3mm2/s,malignant tumor(1.146 ± 0.191)×10-3mm2/s,(0.584 ± 0.139)×10-3mm2/s and mastitis(1.062 ± 0.231)×10-3mm2/s,(0.567 ± 0.115)×10-3mm2/s.The ADCfastvalue of malignant group(4.445 ± 1.616)×10-3mm2/s was significantly higher than that of other groups,and the difference was statistically significant.The ffastvalue of malignant group 0.364 ± 0.148 was significantly lower than that of benign group,normal gland and cyst.ADCfastvalue in the inflammatory group(3.321 ± 0.660)× 10-3mm2/s was significantly lower than that in the malignant group,and the difference was statistically significant.There was no significant difference in ADC,ADCslowand ffast.Conclusion Compared with the single b-value DWI,the parameters of multiple b-value DWI reflected the pathological features of different breast lesions better.The value of ADCfasthas higher diagnostic value in the identification of mastitis and breast cancer.

Objective To investigate the effect of a three-tier delirium care management process in patients with acute stroke in neurology intensive care unit(NICU).Methods A total of 50 patients with acute stroke admitted to the NICU of the Fourth Hospital of Changsha from May to September 2021 were assigned to the control group.The patients in the control group received routine NICU nursing care to prevent delirium.Another 50 patients with acute stroke admitted to the NICU from December 2021 to April 2022 were assigned to the trial group.They were managed with the three-tier delirium nursing management process on top of the routine NICU nursing care for the control group.The incidence of ICU delirium(DICU),duration of DICU,length of stay in NICU and the incidence of delirium-related adverse events were compared between the two groups.The degree of delirium and cognitive function before and after the intervention were compared between the two groups as well.Results The trial group had significantly shorter duration of DICU and NICU stay(both P<0.05)and lower incidence rate of delirium-related adverse events(P<0.05)compared to the control group.After the intervention,the trial group showed significantly lower scores on the intensive care delirium screening checklist(ICDSC)and significantly higher scores of cognitive function compared to those of the control group(both P<0.05).Conclusion The three-tier delirium nursing management process can lower the occurrence of delirium in NICU patients with acute stroke,shorten the NICU stay,reduce the safety risk in nursing,and improve the cognitive function.