Objective To investigate the location of receptor interacting protein 3( RIP3) in Necroptosis and its function in this signal passage, and explore the relationship between receptor interacting protein 1 ( RIP1 ) and RIP3 in nuclear translocation. Methods Primary cerebrocortical neurons were cultured for 12 days,then pre-treated with zVAD-fmk(20μ,mol/L) for half an hour to block apoptosis. ①Extracting nuclear and cytoplasmic protein after neurons were exposed to TNF for different time ,then protein levels of RIP3 were analyzed by western blot and immunofluorescence for qualitative observation;②In the following research,the neurons were treated with Nec-1 and shRlPl ,then the protein level of RIP1 and RIP3 with western blot were analyzed, cell viability were determined by measuring LDH levels. Results ①In signaling pathways of necroptosis, the protein level of RIP3 in cytoplasmic decreased gradually with prolonged TNF exposure, to the corresponding it rolled up in nucleus and a-chieved the peak in 12 hours of TNF treatment ( Cytoplasmic 0. 45 ± 0. 03 ,0. 41 ± 0. 02,0. 73 ± 0. 03 ,0. 90 ± 0.01,1.15 ±0.04,1.30 ±0.02,0.99 ±0.03,0.63 ±0. 03;Nucleus 0. 07 ±0.02,0. 26 ±0.02,0. 57 ±0. 02,0. 68 ± 0.02,0. 80 ± 0.01,0.92 ± 0.02,1.28 ± 0.03,0. 87 ± 0.02) (P < 0.01). ②Blocking the relationship between RIP1 and RIP3 with necrostatin-1 and shRIPl , nuclear translocation of RIP3 decreased and caused a great increase in cell viability( 1.00 ±0.05,0.39 ±0.03,0.50 ±0. 03) (P<0. 01). Conclusion RIP3 mainly locates in cy-tolymph of normal cells,it translocates into nucelus as necroptosis takes place. RIP1 function with RIP3 in nuclear translocation. Block nuclear translocation of RIP3 is a potential way to protect cells.

Objective To explore the features and influencing factors of ambulatory blood pressure in chronic kidney disease (CKD) patients.Methods A total of 540 CKD patients from May 2010 to May 2012 in our department were enrolled in this study.Ambulatory blood pressure monitoring was carried out.Blood pressure (BP),proteinuria and other clinical parameters were measured regularly.Ultrasonography was used to evaluate cardiac structure and function,carotid intima-media thickness and plaque.Univariate and multivariate analysis were used to examine the association between BP and clinical parameters.Results 63.9％ of CKD patients was non-dipper BP pattern,and 36.1％ was dipper BP pattern.As compared to dipper BP patients,those with non-dipper BP had higher ratio of nighttime/daytime proteinuria (0.51±0.29 vs 0.42±0.21,P ＜ 0.01),lower estimated glomerular filtration rate (eGFR) [(56.2±48.2) vs (75.5±56.5) ml· min 1 · (1.73 m2)-1,P ＜ 0.01],higher serum cystatin C[(2.8±2.0) mg/L vs (2.1±2.0) mg/L,P ＜ 0.01],higher left ventricular mass index [(53.7±23.2) vs (45.1± 16.3) g/m2,P ＜ 0.01] and severely damaged left ventricular diastolic function and higher carotid intima-media thickness [(0.7±0.3) vs (0.6±0.2) mm,P＜ 0.01].Nighttime blood pressure was independent predictor for proteinuria,eGFR and left ventricular mass index.Conclusions Nondipper blood pressure pattern is very common in CKD patients.Nighttime pressure is closely associated to renal damage and cardiovascular injuries.

Objective To explore whether hypoxia-inducible factor-1α( HIF-1α )is involved in oxygenglucose deprivation (OGD)induced caspase-independent cell death in primary cortical neurons and whether their expression is infected by necrostatin-1 ( Nec-1 ).Methods ( 1 ) Primary cerebrocortical neurons were cultured for 14 days.Pretred z-VAD.Fmk (z-VAD)and Nec-1 with 0.1,1,5,10,25 and 50 μ mol/L before the neurons were exposed to OGD for 2 hours and reoxygenated for 12 hours,then cell viability was determined by measure LDH level.(2)Pretred z-VAD before the neurons were exposed to OGD for 2 hours,then reoxygenated for 0,2,6,12,24and 48 hours.Then western blot analysis protein level of HIF-1 α ;rt-PCR check its RNA level.(3)Pretred z-VAD and Nec-1 with 25μ mol/L before the neurons were exposed to OGD for 2 hours and reoxygenated for 12 hours.Then western blot analysis protein level of HIF-1 α; rt-PCR check its RNA level.Result ( 1 ) When cells were pretread Nec-1 with 5 μ mol/L(6.97 ± 0.06),the level of LDH was lower than cells untreated( 14.23 ± 0.08 ) (P＜ 0.05);At 25 μmol/L( 2.21 ± 0.05),the level of LDH was essentially the same as that of the control( 1.03 ±0.03 ) (P＞0.05).(2)The protein level of HIF-1 αwas different from normal (0.24 ±0.01 ) when exposed to OGD for 2 hours and reoxygenated for 2 hours (0.57 ± 0.09) and was highest after cells were exposed to OGD for 2 hours and reoxygenated for 12 hours(0.91 ± 0.08 ) (P＜ 0.05 ).The RNA level of HIF-1 α when cells were exposed to OGD was not deferent from normal (P ＞ 0.05 ).( 3 ) When cells were pretread with Nec-1 (0.32 ± 0.04 ),the protein level of HIF-1α were lower than untreated(0.83 ±0.03) (P＜0.05),but the RNA level of HIF-1α had no deference(P ＞ 0.05).Conclusion HIF-1α was involved in cell' s caspase-independent cell death;Nec-1 can protect neurons through inhibiting the expression of HIF-1α.

Objective To evaluate the diagnostic value of the metabolic parameters for differentiating focal autoimmune pancreatitis (F-AIP) and pancreatic cancer (PC) by dual time 18F-FDG PET/CT scan.Methods Ten F-AIP patients and 20 PC patients in Changhai Hospital from May 2011 to November 2014 were enrolled in this study.All the AIP patients were histological confirmed or diagnosed by clinical follow up.The PC patients were histological confirmed and gender-and age-matched with F-AIP patients.50％ SUVmax was set as the threshold to fine-tune the boundary of interest.The extracted parameters included SUV SUV metabolic tumor volume (MTV),total lesion glycolysis (TLG),target-to-background ratio (TBR) and the retention indexes(RI) of all the parameters above.The PET/CT imaging features were also observed.Results The high metabolic lesions were observed in both F-AIP patients and PC patients.There were 6 F-AIP patients whose lesion was located in pancreas head,4 F-AIP patients whose lesion was located in pancreas body and tail.There were 12 PC patients whose lesion was located in pancreas head,8 PC patients whose lesion was located in pancreas body and tail.In F-AIP patients,2 cases had dilated pancreatic duct,6 had dilated biliary duct,8 had increased metabolism in mediastinal lymph node and 2 had abdominal lymphadenopathy,which were 8,5,5 and 14 cases in PC patients.The positive rate of mdeiastinal lymphadenopathy in F-AIP patients was statistically higher than that in PC patients,while the positivity rate of abdominal lymphadenopathy in AIP patients was lower than that in PC patients.The difference was statistically significant (both P ＜ 0.05).There were no statistical differences on the positivity rate of the dilated pancreatic duct,intra-and extra-hepatic bile duct between two groups.SUVmax,SUVmean and MTV in F-AIP were 5.37 ± 0.88,3.48 ± 0.66,21.79 ±15.60 in early stage and 6.45 ±1.51,4.23 ± 1.10,19.36 ± 14.63 in delayed stage,and those in PC were 8.31 ±3.08,5.41±1.95,9.26±8.35 in early stage,and 9.75±3.86,6.36±2.56,9.09±10.71 in delayed stage.SUVmax and SUVmean in F-AIP were lower than those in PC,whereas MTV were larger in F-AIP than that in PC.ROC curves for SUVmax,SUVmean and MTV were made.The AUC of SUV was the highest at 0.85,the cut-off value was 4.45,the corresponding sensitivity was 65％ and the specificity was 90％.TLG,TBR and RI of all the parameters were not statistically different in F-AIP and PC.Conclusions The 18F-FDG PET/CT metabolic parameters,such as SUVmax,SUVmean,MTV,could be of special diagnostic significance in discriminating F-AIP from PC.

Objective To investigate the effect of overexpression of miR-30b on the proliferation,cell cycle,apoptosis and invasion of gastric cancer cell line SGC-7901 and AGS,and the inhibitory effect on the tumor formation in vivo.Methods SGC-7901 and AGS cells were transfected with miR-30b mimics and miR-control,and qRT-PCR was used to detect the expression levels of miR-30b.Western blot assay was used to detect the expression of eIFSA2 protein.CCK-8 assay was used to measure the cell proliferation.Flow cytometry was used to analyze cell cycle and apoptosis.Transwell assay was used to detect cell invasion.In addition,the SGC-7901 and AGS cells transfected with miR-30b mimics and miR-control were injected into nude mice to observe the tumor formation and the expression of eIFSA2 protein in vivo.Results Results of qRT-PCR showed that the relative expression of miR-30b was significantly higher than that of miR-control group (P ＜ 0.05).Western blot assay showed that the expression of eIF5A2 protein was decreased in miR-30b mimics group.CCK-8 assay showed that cell proliferation was inhibited in miR-30b mimics group.The result of flow cytometry showed that the cell cycle decreased and the apoptosis increased in miR-30b group.Transwell assay showed that the cell invasion was significantly lower in miR-30b group than that of control group (P ＜ 0.05).Overexpression of miR-30b inhibited the formation of tumor and decreased the expression of eIF5A2 protein in vivo.Conclusion Overexpression of miR-30b inhibits the proliferation,invasion and tumor formation of gastric cancer cells,and reduces the expression of eIF5A2 protein,which provides a potential target for gastric cancer treatment.

Objective:To compare the effect of systematic nursing in operating room and routine nursing in reducing intraoperative stress injury in patients with spinal fracture and spinal cord injury.Methods:A retrospective case-control study was conducted to analyze the clinical data of 285 patients with cervical or thoracolumbar fracture associated with spinal cord injury admitted to Honghui Hospital Affiliated to Xi'an Jiaotong University from January 2018 to December 2019, including 168 males and 127 females, with the age of 38-59 years [(47.8±8.5)years]. All patients underwent posterior decompression and fusion with internal fixation. Of all, 138 patients received systematic nursing in operating room including systematic evaluation and management before, during and after operation (observation group), and 147 patients received routine nursing including only intraoperative preventive care of pressure ulcer (control group). The incidence of pressure injury on the day after operation, degree of injury and location of injury at postoperative 3 days, and area of injury on the day after operation and at postoperative 3 days were compared between the two groups. The degree of injury was evaluated using the new stress injury staging assessment published by the American National Pressure Ulcer Advisory Panel (NPUAP).Results:The incidence of pressure injury in observation group [5.1% (7/138)] was lower than that in control group [12.2% (18/147)] on the day after operation ( P<0.05). The incidence of stage I, stage II, and stage III pressure injury in observation group [2.9% (4/138), 2.2% (3/138), 0.0%] was also lower than that in control group [8.2% (12/147), 3.4% (5/147), 0.6% (1/147)] at postoperative 3 days ( P<0.05). There was no significant difference in the incidence of pressure injury in the knee, chest, face and anterior superior iliac spine between the two groups ( P>0.05). The total proportion of pressure injury in the face and anterior superior iliac spine was 71% (5/7) in observation group, showing no significant difference from that in control group [83% (15/18)] ( P>0.05). The area of injury was (3.2±1.2)cm 2 and (3.2±1.1)cm 2 in observation group on the day after operation and at postoperative 3 days, lower than that in in control group [(5.1±1.5)cm 2 and (5.1±1.4)cm 2] ( P<0.01). Conclusion:Compared with the routine nursing, systematic nursing in operating room can significantly reduce the incidence, degree and area of intraoperative pressure injury in patients with spinal fracture accompanied by spinal cord injury, and deserves clinical promotion.

Objective:To explore the current situation and influencing factors of quality of life of septic patients in intensive care unit (ICU) after discharge, and to provide theoretical basis for clinical early psychological intervention and continuity of care.Methods:A prospective observational study was conducted. The septic patients who were hospitalized in the department of critical care medicine of the Affiliated Hospital of Jining Medical University and discharged with improvement from January 1 to December 31, 2022 were selected as the research objects. The demographic information, basic diseases, infection site, vital signs at ICU admission, severity scores of the condition within 24 hours after ICU admission, various biochemical indexes, treatment process, and prognostic indexes of all the patients were recorded. All patients were assessed by questionnaire at 3 months of discharge using the 36-item short-form health survey scale (SF-36 scale), the activities of daily living scale (ADL scale), and the Montreal cognitive assessment scale (MoCA scale). Multiple linear regression was used to analyze the factors influencing the quality of life of septic patients after discharge from the hospital.Results:A total of 200 septic patients were discharged with improvement and followed up at 3 months of discharge, of which 150 completed the questionnaire. Of the 150 patients, 57 had sepsis and 93 had septic shock. The total SF-36 scale score of septic patients at 3 months of discharge was 81.4±23.0, and the scores of dimensions were, in descending order, role-emotional (83.4±23.0), mental health (82.9±23.6), bodily pain (82.8±23.3), vitality (81.6±23.2), physical function (81.4±23.5), general health (81.1±23.3), role-physical (79.5±27.0), and social function (78.8±25.2). There was no statistically significant difference in the total SF-36 scale score between the patients with sepsis and septic shock (82.6±22.0 vs. 80.7±23.6, P > 0.05). Incorporating the statistically significant indicators from linear univariate analysis into multiple linear regression analysis, and the results showed that the factors influencing the quality of life of septic patients at 3 months after discharge included ADL scale score at 3 months after discharge [ β= 0.741, 95% confidence interval (95% CI) was 0.606 to 0.791, P < 0.001], length of ICU stay ( β= -0.209, 95% CI was -0.733 to -0.208, P = 0.001), duration of mechanical ventilation ( β= 0.147, 95% CI was 0.122 to 0.978, P = 0.012), total dosage of norepinephrine ( β= -0.111, 95% CI was -0.044 to -0.002, P = 0.028), mean arterial pressure (MAP) at ICU admission ( β= -0.102, 95% CI was -0.203 to -0.007, P = 0.036) and body weight ( β= 0.097, 95% CI was 0.005 to 0.345, P = 0.044). Conclusions:The quality of life of patients with sepsis at 3 months after discharge is at a moderately high level. The influencing factors of the quality of life of patients with sepsis at 3 months after discharge include the ADL scale score at 3 months after discharge, the length of ICU stay, the duration of mechanical ventilation, the total dosage of norepinephrine, MAP at ICU admission and body weight, and healthcare professionals should enhance the treatment and care of the patients during their hospitalization based on the above influencing factors, and pay attention to early psychological intervention and continued care for such patients.

The identification of clinical questions for clinical practice guidelines of Chinese patent medicine （CPM） is important for subsequent evidence retrieval， evaluation of evidence quality， formation of recommendations. This paper described a methodological proposal for the identification of clinical questions for CPM guidelines to highlight the characteristics of Chinese patent medicine and reflect its effect in specific stage of the disease. Considering four aspects， namely， the drug of Chinese patent medicine （D）， the specific disease stage （S）， comparison （C）， and specific outcome （O）， DSCO framework has been proposed to formulate the clinical questions. Multi-source information through scientific research， policy or standard documents， and clinical data are suggested for collecting clinical questions， and clear selection criteria should be set to finalize the clinical questions to be addressed by the guideline. In addition， the above process needs to be transparently and publicly reported in order to ensure the clarity and completeness of the guidelines.

OBJECTIVEUsing simultaneous multi-gene mutation screening to investigate the new method molecular epidemiological basis of 225 patients with nonsyndromic hearing loss in Tianjin, and verifying the for simultaneous multi-gene mutation screening.

METHODSTwo hundred and twenty-five patients with severe non-syndromic deafness from Tianjin CDPF and Association of the Deaf were included in the study. The single nucleotide polymorphisms scan, (SNPscan) technique was used for screening the 115 spots mutations in three common deafness-related genes (GJB2, SLC26A4, mtDNA 12S rRNA) of patients with nonsyndromic hearing loss in Tianjin. We verified the results by Sanger sequencing.

RESULTSAmong the 225 patients, there were 111 cases of deafness caused by mutation (49.3%). Using this method, up to 50% of the patients in our study were identified to have hereditary HL caused by mutations in the three genes. 56 patients with the GJB2 mutations were detected (24.9%), including 30 cases of homozygous mutations (13.3%), 26 patients (11.6%) of compound heterozygous mutations, and 21 cases (9.33%) of single heterozygous mutations. 50 patients with the SLC26A4 mutations were detected (22.2%), including 22 cases of homozygous mutations(9.8%), 28 patients (12.4%) of compound heterozygous mutations, and 22 cases (9.8%) of single heterozygous mutations. mtDNA 12S rRNA A1555G mutation was detected in 5 patients (2.2%). mtDNA 12S rRNA 1494C>T mutation was not detected. We verified the results by Sanger sequencing. The accuracy of the sequencing results was 100%. The SNPscan cost eight hours and 160 yuan (each sample).

CONCLUSIONSApplying SNPscan technology can be accurate, rapid and cost-effective diagnostic screening in patients with hearing loss for etiology investigation. It is expected to become an effective means of large-scale genetic testing for hereditary deafness.

Connexin 26 ; Connexins ; genetics ; DNA Mutational Analysis ; methods ; DNA, Mitochondrial ; genetics ; Deafness ; genetics ; Genetic Testing ; methods ; Heterozygote ; Homozygote ; Humans ; Membrane Transport Proteins ; genetics ; Mutation ; Polymorphism, Single Nucleotide ; RNA, Ribosomal ; genetics