OBJECTIVE To investigate the apoptosis-inducing effect of esculetin （Esc） on acute myeloid leukemia （AML） HL-60 cells by regulating the protein kinase B （AKT）/S-phase kinase-associated protein 2 （SKP2）/MutT homolog 1 （MTH1） pathway. METHODS AML HL-60 cells were randomly divided into control group （routine culture）， Esc low-concentration group （L-Esc group， 25 μmol/L Esc）， Esc medium-concentration group （M-Esc group， 50 μmol/L Esc）， Esc high-concentration group （H-Esc group， 100 μmol/L Esc）， and high-concentration of Esc+ SC79 （AKT agonist） group （100 μmol/L Esc+5 μmol/L SC79）. Cell proliferation in each group was detected by MTT assay and colony formation assay. The level of reactive oxygen species （ROS） in cells was measured by using the CM-H2DCFDA fluorescent probe. Cell apoptosis was analyzed by flow cytometry. Western blot assay was performed to detect the expression levels of apoptosis-related proteins [B-cell lymphoma 2 （Bcl-2）， Bcl-2-associated X protein （Bax）， cleaved caspase-3]， AKT/SKP2/MTH1 pathway-related proteins （p-AKT， AKT， SKP2， MTH1）， along with the upstream and downstream proteins of AKT phosphatidylinositol 3-kinase （PI3K）， cyclin-dependent kinase inhibitor 1 （P21） and cyclin-dependent kinase inhibitor 1B （P27）. RESULTS Compared with control group， the cell viability， colony number， and the phosphorylation levels of AKT and PI3K proteins as well as protein expressions of SKP2， MTH1 and Bcl-2 were significantly decreased （P＜0.05）， while ROS level， apoptosis rate， and the expression levels of Bax， cleaved caspase-3， P21 and P27 proteins were significantly increased （P＜0.05）. Moreover， the effects of Esc exhibited concentration-dependence （P＜0.05）. Compared with H-Esc group， above indexes of high-concentration of Esc+ SC79 group were reversed significantly （P＜0.05）. CONCLUSIONS Esc may promote massive ROS production and induce activation of apoptosis in HL-60 cells by inhibiting the AKT/SKP2/MTH1 pathway， thus inhibiting the proliferation of HL-60 cells.

Objective:To investigate the risk factors associated with one-year mortality following surgery for intertrochanteric fractures in elderly patients and develop a survival prediction model.Methods:A retrospective analysis was conducted on clinical data from 532 elderly patients with intertrochanteric fractures admitted to the People's Hospital of Xinjiang Uygur Autonomous Region and the People's Hospital of Xinyang between January 2020 and September 2022.Patient demographics, laboratory indicators, and surgical variables were documented.The primary outcome assessed was the one-year mortality rate.Risk factors were identified through univariate and multivariate Cox regression analyses, leading to the development of a prognostic model.The model's predictive performance was evaluated using the Concordance Index(C-Index), time-dependent receiver operating characteristic(ROC)curve, calibration curve, and decision curve analysis(DCA).Results:Multivariate Cox regression analysis identified several key factors associated with one-year mortality after intertrochanteric fractures in elderly patients.These factors included the modified five-item frailty index( OR=1.338, 95% CI: 1.147-1.561, P<0.001), ICU admission( OR=1.694, 95% CI: 1.230-2.333, P=0.001), preoperative hemoglobin levels( OR=1.281, 95% CI: 1.016-1.616, P=0.036), surgical waiting time( OR=1.570, 95% CI: 1.063-2.319, P=0.023), and age( OR=2.196, 95% CI: 1.712-2.816, P<0.001).The prediction model showed good consistency with a C-Index of 0.769(95% CI: 0.723-0.818)in the modeling group and 0.715(95% CI: 0.612-0.750)in the validation group.Time-dependent ROC areas under the curve were 0.802(95% CI: 0.722-0.850)and 0.718(95% CI: 0.640-0.808)for the modeling and validation groups, respectively.Calibration curves for both groups indicated a good model fit, and decision curve analysis demonstrated a positive net benefit, highlighting the clinical applicability of the model. Conclusions:The modified five-item frailty index, ICU admission, preoperative hemoglobin, surgical waiting time, and age independently predict one-year mortality after surgery for intertrochanteric fractures in elderly patients.This prognostic model, utilizing these factors, shows high predictive accuracy, assisting clinicians in quick personalized assessments and setting informed expectations in clinical practice.

Irinotecan is an anticancer topoisomerase I inhibitor that acts as a prodrug of the active metabolite, SN-38. Unfortunately, the limited utility of irinotecan is attributed to its pH-dependent stability, short half-life and dose-limiting toxicity. To address this problem, a novel trivalent PEGylated prodrug (PEG-[Irinotecan]₃) has been synthesized and its full-profile pharmacokinetics, antitumor activity and toxicity compared with those of irinotecan. The results show that after intravenous administration to rats, PEG-[Irinotecan]₃ undergoes stepwise loss of irinotecan to form PEG-[Irinotecan]_3‒x (x = 1,2) and PEG-[linker] during which time the released irinotecan undergoes conversion to SN-38. As compared with conventional irinotecan, PEG-[Irinotecan]₃ displays extended release of irinotecan and efficient formation of SN-38 with significantly improved AUC and half-life. In a colorectal cancer-bearing model in nude mice, the tumor concentrations of irinotecan and SN-38 produced by PEG-[Irinotecan]₃ were respectively 86.2 and 2293 times higher at 48 h than produced by irinotecan. In summary, PEG-[Irinotecan]₃ displays superior pharmacokinetic characteristics and antitumor activity with lower toxicity than irinotecan. This supports the view that PEG-[Irinotecan]₃ is a superior anticancer drug to irinotecan and it has entered the phase II trial stage.

Objective: To carry out a investigation on molecular epidemiological features of tick-borne Brucella in Xinjiang Uygur Autonomous Region (Xinjiang), and to provide a scientific basis for formulation of effective preventive and control measures. Methods: In 2016-2018, parasitic ticks (including engorged females) were collected on the body surface of livestock in 10 counties (cities) along the border of Xinjiang. The free-living ticks were collected by flagging method in Alashankou. The engorged female was placed in a breathable insect tube for spawning, each egg batch was divided into two parts: one part was tick eggs, while the second part was allowed further larval development. All ticks were identified by molecular biology (16S rRNA) identification. Tick DNA was extracted, PCR was performed based on Brucella omp22 and IS711, and amplification products were sequenced and analyzed by BLAST. Results: A total of 1 084 ticks were collected in 11 counties (cities), of them 747 were parasitic ticks (including 34 engorged females) and 337 were free-living ticks. Based on 16S rRNA identification, 1 084 ticks belonged to 4 genera and 5 species, and the proportions of Dermacentor nuttalli, Dermacentor marginatus, Haemaphysalis punctata, Hyalomma asiaticum and Rhipicephalus turanicus were 29.43% (319/1 084), 16.51% (179/1 084), 10.42%(113/1 084), 37.27% (404/1 084), and 6.37% (69/1 084), respectively. A total of 214 Brucella-positive nucleic acid samples were detected, the positive rate was 19.74%. The parasitic ticks' positive rate was 25.30% (189/747), and the free-living ticks' positive rate was 7.42% (25/337), parasitic ticks' positive rate was higher than that of free-living ticks (χ²=46.873, P < 0.05). Two Brucella melitensis nucleic acid samples were detected in 34 "engorged females-tick eggs" developmental stage, and one Brucella melitensis nucleic acid sample was detected in 22 "tick eggs-larvae" developmental stage. Conclusions Brucella is widely distributed in parasitic ticks and free-living ticks in Xinjiang border areas, and the parasitic ticks' positive rate is obviously higher than that of free-living ticks. The Brucella melitensis has potential transovarian transmission and transstadial transmission in ticks. In the prevention and control of livestock brucellosis, the work of killing ticks should be strengthened, including parasitic ticks on the body surface and free-living ticks in the environment.

Objective To explore the miRNA regulating the potential cancer-promoting gene CCL18 in cutaneous malignant melanoma.Methods Bioinformatics analysis was conducted by using online software miRanda and TargetScan,so as to predict the miRNA targeting CCL18 gene.Three kinds of C CL18 3'UTR dual-luciferase reporter vectors,including mutant 3'UTR vector (mutant 3'UTR group),wildtype 3'UTR vector (wild-type 3'UTR group) and empty vector (blank control group),as well as miRNA vectors carring selected miRNAs were constructed according to human gene sequence analysis,and then were used to co-transfect 293T cells.After 48-hour treatment,the cells were lysed for detection of luciferase activity.Real-time fluorescence-based quantitative PCR was performed to measure the expression of CCL 18 and selected miRNA in 14 fresh malignant melanoma tissue specimens and 14 paracancerous normal skin tissue specimens (control tissues),and their correlations were analyzed.Results Online software analysis showed that some miRNAs were identified to target the 3'UTR of CCL18 gene,including miR-183,miR-128 and miR-33a.Luciferase reporter vectors and miRNA vectors were constructed successfully.As luciferase activity assay showed,when miR-183 and miR-128 were bound to the CCL18 3'UTR,the luciferase activities were significantly higher in their mutant 3'UTR groups (11.63 ± 0.42;8.80 ± 0.49) than in their wild-type 3'UTR groups (4.86 ± 0.39;5.01 ± 0.54;both P ＜ 0.05) and blank control groups (2.41 ± 0.13;2.39 ± 0.05;both P ＜ 0.01),while there were no significant differences between miR-33a-hinding mutant 3'UTR group (6.41 ± 0.47) and miR-33a-binding wild-type 3'UTR group (6.16 ± 0.22,P ＞ 0.05).Real-time fluorescence-based quantitative PCR revealed higher mRNA expression of the CCL18 gene (3.52 ± 1.68),but lower expression of miR-183 (0.49 ± 0.32),miR-128 (0.30 ± 0.20) and miR-33a (0.46 ± 0.40) in the malignant melanoma tissues compared with the control tissues.The mRNA expression of the CCL18 gene was negatively correlated with the expression of miR-128 (rs =-0548,P ＜ 0.05),but showed no significant correlations with the expression of miR-183 and miR-33a (both P ＞ 0.05).Conclusion miR-128 may play a role in regulating the potential malignant melanoma-promoting gene CCL18.

BACKGROUND:A short-peptide small molecule hydrogel (SMH) developed in the previous study has more obvious advantages than other hydrogels to improve local microenvironment, carry bioactive substances and interfere with stem cell signal transduction pathways. OBJECTIVE:To explore the effect of SMHs on bone marrow mesenchymal stem cells (BMSCs) proliferation, apoptosis and differentiation into myocardial cells. METHODS: (1) Passage 9 rat BMSCs in vitro were divided into control group and experimental group, followed by routine culture and culture in SMHs, respectively. At 7 days of culture, cell proliferation and apoptosis were detected. Cells in the two groups were exposed to anaerobic environment for 12 hours, and expression levels of Bcl-2, Bax and Caspase-3 in BMSCs were detected. (2) Passage 9 BMSCs were divided into four groups and then cultured in 5-azacytidine, SMHs, SMHs+5-azacytidine, and L-DMEM (normal control), respectively. After 4 weeks of induction, expression of CTnT, desmin and Cx-43 proteins was detected and expression levels of early cardiac transcription factors, NKX2.5 and GATA-4, were also measured. RESULTS AND CONCLUSION: (1) Compared with the control group, better proliferation and lower apoptosis of BMSCs were found in the experimental group. Under anaerobic conditions, the number of survival cells was reduced in both groups, but less apoptosis or necrosis was found in the experimental group than the control group (P < 0.05). Moreover, the level of Bcl-2 was higher in the experimental group than the control group (P < 0.01), while the levels of Bax and Caspases-3 protiens were lower in the experimental group than the control group (P < 0.01). (2) NKx2.5 and GATA-4 mRNA expression was found in both 5-azacytidine and SMHs+5-azacytidine groups, and moreover, the mRNA levels of early cardiac transcription factors were significantly higher in the SMHs+5-azacytidine group than in the 5-azacytidine group (P < 0.05). In the normal control group, cTnT expressed negatively, and desmin and Cx-43 expressed weakly. The expression of cTnT, desmin and Cx-43 proteins was higher in the SMHs+5-azacytidine group than in the 5-azacytidine and SMHs groups, while there was no significant difference between the latter two groups. To conclude, SMHs as a culture medium is conducive to the proliferation of BMSCs, reduces cell apoptosis, and promotes myocardial differentiation of BMSCs.

Objective Using single direction dispersion breathless DWI, to analyze the value of DWI for vertebral bone marrow infiltration in patients with acute leukemia (AL). MethodsForty-two patients with AL and 15 healthy volunteers received vertebral sagittal DWI with single shot spin-echo echoplan imaging (SS-SE-EPI) sequence( b value ＝ 0,650 s/mm2) at a GE Signa Excite 1. 5 T scanner with 8 channels body coil. DWI for all patients were performed from three directions, including from superior to inferior (S/I), from anterior to posterior (A/P) and from right to left (R/L). The apparent diffusion coefficient (ADC) value was measured on ADC map from each direction using GE-Function tool DWI software. Forty two patients consisted of 13 onset with untreated patients and 29 treated patients (7 nonremission,8 complete remission and 14 consolidation therapy). The ADC values among the three diffusion directions were compared. Analysis of variance and t test were used to compare the ADC values in different AL stages, Pearson correlation analysis was used to analyze the correlation between ADC values and the percentage of bone marrow progenitor cells. Results The ADC values from S/I, A/P and R/L of 362 vertebras in the 57 subjects are (0. 758 ±0. 009) × 10-3 mm2/s, (0. 732 ±0. 009) × 10 -3 mm2/s and (0. 758 ±0. 009) × 10 -3 mm2/s, respectively. There is no statistical significance( F ＝ 2. 958, P ＞ 0. 05 ).The ADC values from S/I of 94 vertebras in 15 healthy volunteers is (0. 697 ± 0. 122) × 10 -3 mm2/s, of 85 vertebras in 13 untreated AL patients is (0. 592 ±0. 071 ) × 10-3mm2/s. There is statistical significance between them ( t ＝ 2. 568, P ＜ 0. 05 ) ; The ADC value of 183 vertebras in 29 treated AL patients [ ( 0. 796 ±0. 225 ) × 10-3mm2/s]is higher than that in untreated patients with statistical significance (t ＝ -1. 332,P ＜0. 05). One hundred and forty vertebras in patients with complete remission and consolidation therapy were [ (0. 786 ±0. 184) × 10-3 mm2/s],and 43 vertebras in patients with non-remission(NR) [ (0. 804 ±0. 327 ) × 10 - 3 mm2/s], there was not statistical significance between them ( t ＝ - 0. 160, P ＞ 0. 05 ). The ADC values from S/I direction of untreated patients showed significant negative correlation with the proportion of the blast cell in the bone marrow ( median value 26. 4％. Min 7.9％, Max 48. 2％ ) ( r ＝- 0. 524, P ＜ 0. 05 ). ConclusionsDWI of vertebral bone marrow is isotropy. ADC value is a non-invasive and quantitative index for evaluating the pathogenetic condition of AL.

ObjectiveUsing dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) to evaluate the hemodynamic perfusion characteristics of bone marrow infiltration in patients with acute leukemia (AL). MethodsForty-seven patients with AL received coronal pelvic T1WI DCE-MRI with fast low angle shot (FLASH) sequence. Among them, 25 were initial onset untreated (IOU) patients, 22 were treated AL patients, including 14 with complete remission (CR) and 8 with non-remission ( NR). The hemodynamic perfusion parameters including maximum percentage of enhancement ( Emax ) and slope were determined based on enhancement-time curves ( ETCs ) of iliac and lumbar vertebra. The proportion of marrow myeloblasts was recorded.For all patients, quantitative perfusion parameters of bone marrow infiltration in ilium were compared with those in lumbar. The values of Emax and ES were compared among IOU,CR and NR patients.Correlations between perfusion parameters and histopathological results were assessed. ResultsIn all the 47 patients, the Emax values of bilateral iliac bone marrow ( 15.70 ± 7.06)were slightly higher than that of lumbar bone marrow ( 11. 28 ± 5.52 ), and the difference was statistically significant (P ＜0. 01 ).There was no significant difference in the slop value between bilateral iliac bone marrow (0. 82 ± 0. 12 ) and lumbar bone marrow (0. 80 ± 0. 09 ) ( P ＞ 0. 05 ). In the 25 untreated patients,the Emax and slop values were 17. 15 ± 5.75 and 0. 98 ± 0. 13, respectively; in the 14 CR patients, they were 8. 76 ±3.93 and 0. 26 ± 0. 04, respectively, and in the 8 NR patients, they were 21.62 ± 6. 50 and 1. 38 ± 0. 02, respectively. There was significant difference in the Emax and slop values among the three groups (P＜0. 05).Compared with IOU and NR patients, both the Emax and slop values decreased significantly in iliac bone marrow of AL patients with CR (P ＜ 0. 05 ). There was no significant difference between IOU and NR patients ( P ＞ 0. 05 ). A significant positive correlation was found between Emax value of iliac bone marrow and the proportion of marrow myeloblasts ( r ＝0. 501 ,P ＜0. 05 ). There was a negative correlation between slop value of iliac bone marrow and the proportion of marrow myeloblasts ( r ＝0. 235 ,P ＞0.05).ConclusionsDCE-MRI can beused for evaluating the hemedynamic characteristics of microcirculation of bone marrow infiltration in patients with AL, which can provide useful information in evaluating prognosis and monitoring therapeutic effect.

A mouse-anti-human monoclonal antibody was produced by using the membrane proteins of human lung carcinoma cell line A549 as the immunogen to generate monoclonal antibodies against lung carcinoma with the use of hybridoma techniques. McAb4E7 was prepared successfully. To identify its antigen, proteomic technologies such as two-dimenstional electrophoresis, western blotting and mass spectrometry were employed. The targeting antigen of McAb4E7 expressed positive in human lung cancer cell lines A549 and human hepatocarcinoma cell line HepG2, moreover, the expression of the antigen was stronger in A549 cells. Finally, we obtained one positive protein in A549 cell line that has strong affinity and specificity for McAb4E7, which was identified to be ATP synthase beta subunit. We identified ATP synthase beta subunit as the targeting antigen of lung carcinoma special monoclonal antibody McAb4E7.
Animals ; Antibodies, Monoclonal ; biosynthesis ; chemistry ; immunology ; Antibodies, Neoplasm ; immunology ; Antibody Specificity ; Antigens, Neoplasm ; genetics ; immunology ; Cell Line, Tumor ; Humans ; Lung Neoplasms ; immunology ; Membrane Proteins ; immunology ; Mice ; Mice, Inbred BALB C ; Mitochondrial Proton-Translocating ATPases ; immunology