Objective To investigate the changes of cardiac function and myocardial apoptosis after limb ischemia reperfusion in rabbits.Methods Thirty rabbits were randomly divided into 3 groups:sham-operated control group (group SC),ischemia group (groupⅠ,8 hours of bilateral hind limb ischemia) and ischemia reperfusion group (group IR,4 hours of bilateral hind limb ischemia and 4 hours of reperfusion).Immunohistochemical studies and TUNEL were done to evaluate cell apoptosis.The SOD activity and MDA content in myocardial tissue and plasma LDH activity were determined. The left ventricular function were observed by physiology recording instrument.The cardiac histopathologic changes in experimental rabbits were observed.Results In group IR,the apoptosis index and expression of Bax were significantly increased compared with the other two groups.Similarly,ischemia reperfusion reduced SOD activity and enhanced LDH activity and MDA level,and the left ventricular function decreased in group IR.There was a positive correlation among the MDA content and the LDH activity,the expression of bax,the number of apoptosis cells.Under light microscope, myocardial impairment was found in group IR.Conclusion The myocardial injury occured and the left ventrieular function decreased after limb ischemia reperfusion,and they were associated with oxidative damage and apoptosis.

Objective To explore the characteristic of fractional amplitude of low frequency fluctuation (fALFF) and the relationship with the severity of depression, suicidal ideation and suicide risk in depression patients with suicidal ideation with resting-state functional magnetic resonance imaging (rs-fMRI). Methods Resting state functional magnetic resonance imaging maps were conducted using fractional amplitude of low frequency fluctuation (fALFF) in 52 depression patients (30 with suicidal ideation and 22 without) and 21 healthy controls (HCs). The severity of depression was evaluat-ed by using Hamilton Depression scale(HAMD). The suicidal ideation, the suicide risk in depression patients with sui-cidal ideation were both assessed by the Beck Scale for Suicide Ideation. The correlation between the fALFF value and the score of HAMD and the Beck Scale for Suicide Ideation was analyzed. Results MRI revealed significant differences in fALFF in the left superior/middle occipital gyrus and the right middle/inferior occipital gyrus (P<0.05, AlphaSim cor-rected)between depression patients with suicidal ideation and the HCs. Compared to the HCs, depression patients with-out suicidal ideation showed a higher fALFF in the left middle occipital gyrus (P<0.05, AlphaSim corrected). MRI re-vealed significant differences in fALFF in the left middle occipital gyrus (P<0.01, AlphaSim corrected)and the right mid-dle occipital gyrus (P<0.01, AlphaSim corrected) between depression patients with suicidal ideation and without. The fALFF of left middle occipital gyrus (r=0.366, P=0.046) and right middle occipital gyrus (r=0.513, P=0.004) were posi-tively correlated with the scores of HAMD, respectively whereas were not correlated with suicidal ideation and suicide risk. Conclusions Depression patients with suicidal ideation have an abnormal spontaneous activity in their left and right middle occipital gyrus. The increased activity in these brain areas are probably associated with the severity of de-pression whereas are not associated with suicidal ideation or suicide risk.

Objective To investigate the correlation between the proportion of peripheral blood follicular T helper cells(Tfh cells)and intracellular interleukin-21(IL-21)content with blue light retinal injury.Methods Brown Norway(BN)rats were randomly divided into 4 groups and were exposed to blue light for 3 hours a day to establish retinal light damage model.According to the duration of illumination,the rats was divided into 0 days(control group),3 days(3 d group),7 days(7 d group)and 14 days(14 d group).The proportion of Tfh cells and content of IL-21 in Tfh cells in peripheral blood of each group was detected by flow cytometry and ELISA sepa-rately after illumination.Electroretinogram(ERG)was used to evaluate retinal function.The changes of fundus in rats were observed by fundus photography.The thickness of outer nuclear layer of retina was analyzed by HE staining.Results After retinal blue light injury,with the extension of illumination time,the proportion of Tfh cells in peripheral blood and intracellular IL-21 content both increased(P<0.05).ERG showed that retinal function decreased after light damage and aggravated with the extension of illumination time,the latency and ampli-tudes of A-wave and B-wave increased and decreased respectively(P<0.05).The retinal fundus of rats showed depigmentation in 3 d,and the retinal vessels became thinner and exudate with the extension of illumination time.HE staining showed that the outer nuclear layer of retina(ONL)became thinner(P<0.05).Correlation analy-sis indicated that the proportion of Tfh cells in peripheral blood and the intracellular IL-21 content could jointly reflect the degree of injury(P<0.000 1),and the proportion of Tfh cells in peripheral blood was negatively corre-lated with ONL thickness and the amplitude of a and b waves,positively correlated with the peak time of a and b waves,(P<0.0001).Conclusion The proportion of Tfh cells in peripheral blood and the intracellular IL-21 content were increased after blue light damage to retina,and were significantly increased with the extension of light time with a certain correlation.

Objective:To understand the current state of renal replacement therapy (RRT) in intensive care unit (ICU) of Tianjin public hospital, and to provide scientific evidence and direction for homogenized management and overall level improvement of RRT in Tianjin.Methods:The questionnaires were distributed to the chief or key staff of 33 ICUs from 32 public hospitals in Tianjin by clinical quality control center for critical care medicine of Tianjin and ICU of Tianjin Third Central Hospital. The RRT data of ICUs from January 2020 to December 2021 was collected, including the type and size of ICU, the number of patients undergoing RRT, reasons for initiating RRT, the RRT modes, the anticoagulation and the complications of RRT and so on.Results:A total of 33 valid questionnaires were obtained, with a recovery rate of 100%. The result showed that there were 38 803 patients admitted to the selected ICUs during investigation, and 5 456 of them (14.06%) received RRT. In most ICUs, the reasons of initiating RRT were renal failure, sepsis and volume overload. The mode of RRT was mainly continuous venovenous hemofiltration (CVVH), which was followed by continuous venovenous hemodiafiltration (CVVHDF). Carbonate replacement fluid was the first choice. Heparin was the dominant anticoagulant, and there was an increasing trend in the use of citrate anticoagulation simultaneously. However, heparin-free anticoagulation used mostly in bleeding patients. Overall, the RRT modes and anticoagulation methods were single. Thrombosis was the main iatrogenic factor interrupting RRT in most ICUs, and also the reasons for complications related to catheter or circulation pipeline. It still showed an ineffective anticoagulation of RRT even after increasing the dosage of anticoagulants.Conclusions:RRT is an important organ support method in ICU, which has been widely carried out in ICUs of Tianjin and continues to expand. Despite the positive performance, it still needs to be improved and standarized in some aspects, such as the diversification of RRT modes, anticoagulation, and the complication prevention.

The in vivo assembly of ribosomal subunits is a highly complex process, with a tight coordination between protein assembly and rRNA maturation events, such as folding and processing of rRNA precursors, as well as modifications of selected bases. In the cell, a large number of factors are required to ensure the efficiency and fidelity of subunit production. Here we characterize the immature 30S subunits accumulated in a factor-null Escherichia coli strain (∆rsgA∆rbfA). The immature 30S subunits isolated with varying salt concentrations in the buffer system show interesting differences on both protein composition and structure. Specifically, intermediates derived under the two contrasting salt conditions (high and low) likely reflect two distinctive assembly stages, the relatively early and late stages of the 3' domain assembly, respectively. Detailed structural analysis demonstrates a mechanistic coupling between the maturation of the 5' end of the 17S rRNA and the assembly of the 30S head domain, and attributes a unique role of S5 in coordinating these two events. Furthermore, our structural results likely reveal the location of the unprocessed terminal sequences of the 17S rRNA, and suggest that the maturation events of the 17S rRNA could be employed as quality control mechanisms on subunit production and protein translation.
Cryoelectron Microscopy ; Escherichia coli ; metabolism ; Escherichia coli Proteins ; genetics ; metabolism ; GTP Phosphohydrolases ; genetics ; metabolism ; Mass Spectrometry ; Protein Structure, Secondary ; Protein Structure, Tertiary ; RNA, Ribosomal ; analysis ; metabolism ; Ribosomal Proteins ; chemistry ; genetics ; metabolism ; Ribosome Subunits, Small, Bacterial ; chemistry ; metabolism ; ultrastructure ; Salts ; chemistry

Objective:To evaluate the risk factors of clinical cure and biochemical recurrence (BCR) after radical prostatectomy (RP).Methods:The clinical data of 896 patients who underwent RP at Peking University First Hospital from April 2001 to December 2020 were retrospectively analyzed. Average age was (65.90±6.3) years, median preoperative prostate specific antigen (PSA) was 10.75 (0.36-264.20) ng/ml, median prostate volume was 40.0 (12.0-220.9) ml, median PSA density (PSAD) was 0.27 (0.02-3.42) ng/(ml·g). Clinical staging: 432 cases in T 1c stage, 333 cases in T 2a/bstage, 76 cases in T 2c stage, and 55 cases in ≥T 3 stage. Preoperative Gleason score of biopsy: 193 cases in 3+ 3, 315 cases in 3+ 4, 162 cases in 4+ 3, 226 cases in ≥8. The RP surgery was operated by open or laparoscopic or robot-assisted approach. Clinical cure and BCR were used as the end points for analysis. Clinical cure was defined as a decrease in serum PSA level below 0.03 ng/ml 6 weeks after surgery. BCR was defined as the 2 consecutive serum PSA >0.2ng/ml during the follow-up after RP. Multivariate logistic regression was used to analyze the independent risk factors of clinical cure. The Kaplan-Meier method was used to draw the biochemical recurrence-free survival curve, the log-rank method was used for univariate analysis of BCR, and the Cox regression analysis was used for multivariate analysis. Results:All 896 patients were followed-up for 58 (5-241) months, 678 cases (75.7%) achieved clinical cure. Based on univariate analysis and multivariate analysis, among the preoperative indicators, whether the proportion of positive biopsy needles ≥33% ( P=0.007) and preoperative Gleason score of biopsy ( P=0.041) were independent risk factors of clinical cure. A total of 890 cases were included in the analysis of risk factors of BCR, of whom 172 cases (19.3%) had BCR. The 1-, 5-, and 10-year biochemical recurrence-free survival(BFS)rates were 98.1%, 83.1% and 68.4% respectively. The median BFS has not been reached, and the average BFS was 181 months (95% CI 172-189). The results of univariate and multivariate analysis showed that whether achieved clinical cure ( P=0.001) and postoperative pathological staging ( P<0.001) were independent risk factors of BCR. Conclusions:Whether the proportion of positive biopsy needles≥33% and preoperative Gleason score of biopsy were independent risk factors of clinical cure. Postoperative pathological staging and whether achieved clinical cure may be independent risk factors of BCR.

The human Shwachman-Diamond syndrome (SDS) is an autosomal recessive disease caused by mutations in a highly conserved ribosome assembly factor SBDS. The functional role of SBDS is to cooperate with another assembly factor, elongation factor 1-like (Efl1), to promote the release of eukaryotic initiation factor 6 (eIF6) from the late-stage cytoplasmic 60S precursors. In the present work, we characterized, both biochemically and structurally, the interaction between the 60S subunit and SBDS protein (Sdo1p) from yeast. Our data show that Sdo1p interacts tightly with the mature 60S subunit in vitro through its domain I and II, and is capable of bridging two 60S subunits to form a stable 2:2 dimer. Structural analysis indicates that Sdo1p bind to the ribosomal P-site, in the proximity of uL16 and uL5, and with direct contact to H69 and H38. The dynamic nature of Sdo1p on the 60S subunit, together with its strategic binding position, suggests a surveillance role of Sdo1p in monitoring the conformational maturation of the ribosomal P-site. Altogether, our data support a conformational signal-relay cascade during late-stage 60S maturation, involving uL16, Sdo1p, and Efl1p, which interrogates the functional P-site to control the departure of the anti-association factor eIF6.
Crystallography, X-Ray ; GTP Phosphohydrolases ; chemistry ; metabolism ; Humans ; Protein Domains ; Ribosome Subunits, Large, Eukaryotic ; chemistry ; metabolism ; Saccharomyces cerevisiae ; chemistry ; metabolism ; Saccharomyces cerevisiae Proteins ; chemistry ; metabolism

Objective To analyze the clinical features of patients with coronavirus disease 2019 (COVID-19) in Shanghai and to investigate the risk factors for disease progression to severe cases. Methods The clinical data of 292 adult patients with COVID-19 hospitalized in Shanghai Public Health Clinical Center from January 20, 2020 to February 10, 2020 were retrospectively analyzed, including 21 severe patients and 271 mild patients. The demographic characteristics, epidemiological history, history of underlying diseases and laboratory examinations were compared between the two groups. Measurement data were compared using t test or Mann-Whitney U test. The count data were compared using hi-square test. The binary logistic regression equation was used to analyze the risk factors for the progression of patients to severe cases. Results Among the 292 patients, 21 were severe cases with the rate of 7.2% (21/292). One patient died, and the mortality rate was 4.8% in severe patients. The severe patients aged (65.0±15.7) years old, 19 (90.5%) were male, 11 (52.4%) had underlying diseases, 7 (33.3%) had close relatives diagnosed with COVID-19. The mild patients aged (48.7±15.7) years old, 135 (49.8%) were male, 74 (27.3%) had underlying diseases, 36 (13.3%) had close relatives diagnosed with COVID-19. The differences between two groups were all significant statistically ( t =-4.730, χ 2 =12.930, 5.938 and 4.744, respectively, all P <0.05). Compared with the mild patients, the levels of absolute numbers of neutrophils, alanine aminotransferase, aspartate aminotransferase, lactate dehydrogenase, creatinine, serum cystatin C, C reactive protein (CRP), procalcitonin , D -dimer, pro-B-type natriuretic peptide (proBNP), serum myoglobin, creatine kinase (CK), creatine kinase isoenzyme (CK-MB), serum troponin I (cTnI) in severe patients were all significantly higher ( U =2 091.5, 1 928.0, 1 215.5, 729.0, 1 580.5, 1 375.5, 917.5, 789.5, 1 209.0, 1 434.0, 638.0, 964.5, 1 258.0 and 1 747.5, respectively, all P <0.05), while the levels of lymphocyte count, albumin, transferrin, CD3 + T lymphocyte count, CD8 + T lymphocyte count and CD4 + T lymphocyte count in severe patients were all significantly lower ( U =1 263.5, t =4.716, U =1 214.0, 962.0, 1 167.5 and 988.0, respectively, all P <0.05). Further logistic regression analysis showed that the albumin (odds ratio ( OR )=0.806, 95% CI 0.675-0.961), CRP ( OR =1.016, 95% CI 1.000-1.032), serum myoglobin ( OR =1.010, 95% CI 1.004-1.016), CD3 + T lymphocyte count ( OR =0.996, 95% CI 0.991-1.000) and CD8 + T lymphocyte count ( OR =1.006, 95% CI 1.001-1.010) at admission were independent risk factors for the progression of COVID-19 patients to severe illness (all P <0.05). Conclusions Severe cases of patients with COVID-19 in Shanghai are predominantly elderly men with underlying diseases. Albumin, CRP, serum myoglobin, CD3 + T lymphocyte count and CD8 + T lymphocyte count could be used as early warning indicators for severe cases, which deserve more clinical attention.