Systematical quantitative evaluation of left ventricular mechanical dysfunction in patients with cirrhosis using ultrasonic layer-specific strain imaging / 中华超声影像学杂志

Objective:To assess the left ventricular (LV) myocardial mechanical dysfunction in patients with cirrhosis using ultrasonic layer-specific strain imaging and to explore its value in clinical application.Methods:A total of 80 consecutive cirrhosis patients without cardiovascular diseases were prospectively enrolled from October 2020 to March 2021 in Sichuan Provincial People′s Hospital, 39 of whom were assigned to the compensated group and 41 were assigned to the decompensated group according to the occurrence of portal hypertension. Forty-three healthy volunteers during the same period were randomly recruited as the control group. Transthoracic echocardiography was performed to assess the LV configuration and functional parameters. LV global longitudinal strain in endocardial, middle and epicardial myocardium (GLSendo, GLSmid, GLSepi), and longitudinal strain (LS) in basal, middle and apical segments, and peak strain dispersion (PSD) were obtained using ultrasonic layer-specific strain imaging. ΔLS was calculated by the formula of GLSendo-GLSepi. Then, the differences of related parameters among three groups were compared.Results:①Conventional echocardiography: compared with the control group, the interventricular septum end-diastolic thickness (IVSTd), left ventricular posterior wall end-diastolic thickness (LVPWd), left ventricular mass (LVM) and LVM index (LVMI) were increased in compensated and decompensated groups (all P<0.05), while no significant differences in conventional echocardiographic parameters were identified between the two cirrhosis groups (all P>0.05). ②Global layer-specific strain: compared with the control group, GLSendo, GLSmid, GLSepi and ΔLS were decreased and PSD was increased in compensated and decompensated groups (all P<0.05); Moreover, the decompensated group showed a more impaired GLSendo, GLSmid and GLSepi than compensated group (all P<0.05), whereas there were no significant differences of ΔLS and PSD between the two groups(all P>0.05). ③Segmental layer-specific strain: compared with the control group, LS values of three layers in compensated and decompensated groups were reduced at basal, middle and apical levels (all P<0.05); Compared with the compensated group, LS values of three layers in decompensated group tended to be reduced at above there levels, but only apical segments had significant differences (all P<0.05). Conclusions:There are different degrees of LV mechanical dysfunction in patients with variable severity of cirrhosis. Ultrasonic layer-specific strain imaging has the potential to quantitatively assess the state of cardiac involvement in patients with cirrhosis and to provide visual evidence for the early and accurate diagnosis of myocardial injuries.

Effect of Denticleless E3 Ubiquitin Protein Ligase on Proliferation and Colone Formation of Multiple Myeloma Cells / 中国医学科学院学报

Objective To determine the effect of denticleless E3 ubiquitin protein ligase(DTL)on the proliferation and clone formation of multiple myeloma(MM)cells and investigate the related mechanism. Methods Mononuclear cells were extracted from 34 MM patients.Mononuclear cells harvested from 14 healthy volunteers were used as controls.Quantitative polymerase chain reaction was used to detect the change of DTL at mRNA level.Furthermore,12 MM patients and 2 controls were selected,in whom the change of DTL at protein level was detected by Western blot.Human MM cell line RPMI8226 was divided into control(CON)group and DTL-short hairpin RNA(DTL-shRNA)group,which was infected with the CON and DTL-shRNA virus,respectively,for 48 hours.The infection efficiency was detected by using flow cytometry,the knock-down efficiencies at mRNA and protein levels were detected by quantitative polymerase chain reaction and Western blot,the change of cell counts in the next 0,24,48,72,96 hours were measured with CCK8 assay.The CON and DTL-shRNA cells were cultured in semisolid medium.Ten days later,inverted phase microscopy was used to measure the number of colones that contain more than 50 cells,annexin V/propidium iodide double staining to detect apoptosis,and propidium iodide staning to detect cell cycle.Finally,Western blot was empoyed to detect the phosphorylation of P65 and inhibitory subunit-κBα(IκBα)in nuclear factor-κB(NF-κB)pathway and electrophoretic mobility shift assay(EMSA)to detect the NF-κB transcriptional ability. Results The DTL expression was(1.00±0.12)and(9.36±3.71),respectively in the bone marrow mononuclear cells of healthy volunteers and in the CD138+cells of MM patients(t=3.65,P=0.0024).DTL was also highly expressed in MM CD138+positive cells at protein level.After RPMI8226 was infected by CON and DTL-shRNA virus for 48 hours,green fluorescent protein-positive cells accounted for more than 90%.The relative expression of DTL was(1.00±0.01)and(0.21±0.04)(t=33.19,P<0.0001)at mRNA level and(0.52±0.13)and(0.11±0.02)at protein level(t=5.399,P=0.0057).CCK8 revealed that CON and DTL-shRNA cells proliferated by(1.00±0.03)vs.(1.00±0.02),(2.19±0.28)vs.(1.47±0.13),(3.50±0.14)vs.(2.24±0.19),(5.43±0.41)vs.(3.08±0.14),(7.42±0.17)vs.(4.29±013)after 0,24,48,72,and 96 hours(F=24.58,P=0.001).The number of colone containing more than 50 cells was in 76±4 in CON group and 0 in DTL-shRNA group(P<0.01).The proportion of G1 stage cells was(28.61±8.64)% in CON group and(57.25±10.37)% in DTL-shRNA group(t=3.675,P=0.0213).The proportion of annexin V+in CON and DTL-shRNA groups was(3.21±0.89)% vs.(34.71±18.68)%(t=2.895,P=0.0443).After RPMI8226 was infected with CON or DTL-shRNA virus for 48 hours,the relative expression of phosphorylation P65 was(1.52±0.14)vs.(0.82±0.11)(t=6.81,P=0.0024),the P65 relative expression was(0.25±0.04)vs.(0.24±0.08)(t=0.19,P=0.85),the CON and DTL-shRNA phosphorylation-IκBα relative expression was(0.19±0.03)vs.(0.13±0.02)(t=2.882,P=0.0449),and the IκBα was(0.22±0.05)vs.(1.01±0.06)(t=17.52,P<0.0001).Detection of the transcriptional ability of DTL-shRNA NF-κB by EMSA further confirmed the down-regulation of DTL suppressed the NF-κB transcriptional ability. Conclusions DTL is highly expressed in MM cells,and down-regulation of DTL suppresses the cell proliferation,inhibit the colony formation,and induce cell apoptosis and cell cycle arrest.The effect of DTL on the biological functions of MM cells is related to the change of NF-κB pathway.

Ribosomal Protein S9 Expression in Multiple Myeloma and Its Effect on Cell Biological Function / 中国医学科学院学报

Objective To identify the expression of ribosomal protein S9(RPS9)in multiple myeloma(MM)and explore its effect on the biological characteristics of myeloma cells and the corresponding mechanisms. Methods Bone marrow mononuclear cells were harvested in 10 healthy volunteers(CON group)and bone marrow CD138 +cells from 30 MM patients(CD138+group).Quantitative polymerase chain reaction(qPCR)was performed to detect RPS9 expression at mRNA level.In three cases from CON group and 11 cases from CD138+group,Western blot was performed to detect RPS9 at protein level.GSE19784 dataset was employed to detect the relationships of RPS9 expression with the overall survival rate,nuclear factor-κB(NF-κB),small ubiquitin-like modifier(SUMO),and ubiquitin pathway.After the RPS9 knock-down vector was constructed,flow cytometry was performed to detect the infection efficiency and qPCR and Western blot to detect the knock-down efficiency.RPMI8226 was divided into CON group and RPS9-short hairpin RNA(shRNA)group,in which annexin V allophycocyanin/propidium iodide(PI)double staining was performed to detect the change of apoptosis,CCK8 to detect the proliferation change,and PI staining to detect cell cycle change.After sentrin-specific protease 1(SENP1)overexpression vector was constructed,Western blot was performed to detect the phosphorylation of P65 and inhibitory subunit-κBα(IκBα)from NF-κB pathway in CON,RPS9-shRNA,and RPS9-shRNA-SENP1 cells;in addition,annexin V/PI double staining was also performed to detect the apoptosis in these three cells. Results The relative expression of RPS9 in CON group and CD138+group was(1.00±0.12)and(5.45±0.71),respectively(t=4.291,P=0.0036).Western blot showed RPS9 expression was high in most myeloma CD138+cells.The high expression of RPS9 was associated with both extramedullary invasion and overall survival in GSE19784 dataset.After RPMI8226 was infected with CON or RPS9-shRNA lentivirus for 48 hours,flow cytometry confirmed that the infection efficiencies were above 90% in both groups.qPCR and Western blot confirmed that RPS9 expression was inhibited at both mRNA and protein levels.After RPMI8226 CON and RPS9-shRNA infected with virus for 48 hours,the proportion of annexin V-positive cells in CON and RPS9-shRNA cells was(3.47±0.37)% and(18.60±64.00)%(t=9.015,P=0.0008).The proliferation index significantly differed between CON group and RPS9-shRNA group at 72 hours(t=6.846,P=0.0024).When CON and RPS9-shRNA were infected with virus for 48 hours,the proportion of G2 phase cells was(29.28±3.42)% and(10.43±1.43)%,respectively(t=9.329,P=0.0007).The RPS9 expression was positively correlated with SENP1 in GSE19784 dataset and negatively correlated with IκBα coding gene NFKBIA.Western blot further confirmed that RPS9 knockdown inhibited the expression of SENP1,inhibited the phosphorylation of NF-κB subunit P65 and inhibitor IκBα,and promoted the expression of IκBα.Overexpression of SENP1 not only impeded this effect but also reduced RPS9-induced apoptosis. Conclusions RPS9 is highly expressed in MM CD138+cells and is associated with overall survival and extramedullary infiltration.Inhibition of RPS9 can promote apoptosis,cell cycle arrest,and proliferation of myeloma cells.RPS9 can affect the activation of NF-κB pathway and cell apoptosis through SENP1,suggesting that SENP1 may be a key factor in the biological effect of RPS9.

The epidemiological characteristics and the source of infection of reemerge human rabies from 2012 to 2017 in Qinghai / 中华疾病控制杂志

Objective To understand the epidemic situation and the source of infection of the reemerge human rabies in Qinghai. Methods We collected the data on human rabies and the data on the cases of multi- victims bitten by the identical dog, and also the laboratory data of the nucleoprotein ( N) gene of rabies virus from the samples which were detected by reverse transcription-polymerase chain reaction (RT-PCR) and direct immunofluorescence assay (DFA) from 2012 to 2017, to describe the epidemiological characteristics of human rabies and the prevalence of rabies virus in host animals, and to explore the source of infection of reemerge human rabies. Results A total of 7 human cases were reported in 2012-2017 in Qinghai province, among which 1 was bitted by wolf, 2 were bitted by stray dogs, 3 were bitted by domestic dogs which injured by stray dogs or wolfs. A total of 892 canine brain tissue samples were collected, from which 46 positive samples were detected with the positive rate of 5.16% (95% CI：3.70%-6.61%). The positive samples were collected from the nomadic region, which were consistent had the location of the human rabies. The samples collected from the cases of multi-victims bitten by the identical dog/animal had the positive rate of 73.08%, and 4 out of 7 human rabies were exposed to the cases of multi-victims bitten by the identical dog/animal. Genetic sequencing of the rabies virus detected from canine brain tissue samples were belong to China IV lineage, which was closely related to the Arctic clade. Conclusions The reemerging rabies happened in nomadic region of Qinghai province could be a consequence of spillover from wildlife especially from wolfs. The better surveillance system covering the human, livestock and wildlife should be set up to mitigate the rabies virus spread from the wildlife.

Analysis of risk factors of infection for complex tibial plateau fractures after operation / 中国骨伤

<p><b>OBJECTIVE</b>To analyze the risk factor of infection for complex tibial plateau fractures after operation.</p><p><b>METHODS</b>Totally clinical data of 293 patients with complex tibial plateau fractures underwent open reduction and internal fixation were retrospectively analyzed from September 2010 to March 2015, including 199 males and 94 females, ranging in age from 17 to 80 years old with an average of 47.3 years old. The possible risk factors such as gender, age, smoking, diabetes, type of fracture(open/closed), classification of open fracture(Gustilo-Anderson classification), classification of soft tissue injury in closed fracture (Tscherne-Gotzen classification), fracture classification(Schatzker V/VI), osteofascial compartment syndrome, ASA score, anesthesia, timing of surgery, operative time(<=150 min/>150 min), surgical approach, combined approach or not, internal fixation site were studied. The multivariate Logistic regression model was used to analyze the risk factors.</p><p><b>RESULTS</b>Twelve patients were infected of all 293 patients after operation, the infection rate was 4.10%. Univariate analysis showed that fracture type(χ ² =14.496,=0.001), fracture classification(χ²=4.560,=0.033), osteofascial compartment syndrome(χ²=15.631,=0.001), operative time(χ²=11.233,=0.001) were correlated with complex tibial plateau fractures postoperative infection. Multivariate analysis showed that open fractures(χ²=9.696,=0.002) and osteofascial compartment syndrome(χ²=9.119,=0.003) were complex tibial plateau fracture risk factors for infection after operation.</p><p><b>CONCLUSIONS</b>Open fractures and osteofascial compartment syndrome are risk factor of complex tibial plateau fracture for infection after operation. While through debridement for open fracture patients, early diagnosis and promt treatment for osteofascial compartment syndrome could reduce incidence of infection.</p>