Design, docking optimization, and evaluation of biotin-PEG4-1,8-naphthalimide as a potent and safe antitumor agent with dual targeting of ferroptosis and DNA.

A set of biotin-polyethylene glycol (PEG)-naphthalimide derivatives 4a-4h with dual targeting of ferroptosis and DNA were designed and optimized using docking simulation as antitumor agents. Docking simulation optimization results indicated that biotin-PEG4-piperazine-1,8-naphthalimide 4d should be the best candidate among these designed compounds 4a-4h, and therefore, we synthesized and evaluated it as a novel antitumor agent. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and MGC-803 and U251 xenograft models identified 4d as a good candidate antitumor agent with potent efficacy and safety profiles, compared with amonafide and temozolomide. The findings of the docking simulations, fluorescence intercalator displacement (FID), western blot, comet, 5-ethynyl-2'-deoxyuridine (EdU), flow cytometry, transmission electron microscopy, and BODIPY-581/591-C11, FerroOrange, and dihydroethidium (DHE) fluorescent probe assays revealed that 4d could induce DNA damage, affect DNA synthesis, and cause cell cycle arrest in the S phase in MGC-803 cells. Also, it could induce lipid peroxidation and thus lead to ferroptosis in MGC-803 cells, indicating that it mainly exerted antitumor effects through dual targeting of ferroptosis and DNA. These results suggested that it was feasible to design, optimize using docking simulation, and evaluate the potency and safety of biotin-PEG-1,8-naphthalimide as a antitumor agent with dual targeting of ferroptosis and DNA, based on a multi-target drug strategy.

[Association between maternal gestational diabetes mellitus and the risk of autism spectrum disorder in offspring]. / æ¯äº²å¦Šå¨ ç³–å°¿ç— ä¸Žå­ä»£å­¤ç‹¬ç—‡è°±ç³»éšœç¢å‘ç”Ÿé£Žé™©çš„å ³è”åˆ†æž.

OBJECTIVES: To explore the association between maternal gestational diabetes mellitus (GDM) exposure and the development of autism spectrum disorder (ASD) in offspring. METHODS: A case-control study was conducted, recruiting 221 children with ASD and 400 healthy children as controls. Questionnaires and interviews were used to collect information on general characteristics of the children, socio-economic characteristics of the family, maternal pregnancy history, and maternal disease exposure during pregnancy. Multivariate logistic regression analysis was used to investigate the association between maternal GDM exposure and the development of ASD in offspring. The potential interaction between offspring gender and maternal GDM exposure on the development of ASD in offspring was explored. RESULTS: The proportion of maternal GDM was significantly higher in the ASD group compared to the control group (16.3% vs 9.4%, P=0.014). After adjusting for variables such as gender, gestational age, mode of delivery, parity, and maternal education level, maternal GDM exposure was a risk factor for ASD in offspring (OR=2.18, 95%CI: 1.04-4.54, P=0.038). On the basis of adjusting the above variables, after further adjusting the variables including prenatal intake of multivitamins, folic acid intake in the first three months of pregnancy, and assisted reproduction the result trend did not change, but no statistical significance was observed (OR=1.94, 95%CI: 0.74-5.11, P=0.183). There was an interaction between maternal GDM exposure and offspring gender on the development of ASD in offspring (P<0.001). Gender stratified analysis showed that only in male offspring of mothers with GDM, the risk of ASD was significantly increased (OR=3.67, 95%CI: 1.16-11.65, P=0.027). CONCLUSIONS: Maternal GDM exposure might increase the risk of ASD in offspring. There is an interaction between GDM exposure and offspring gender in the development of ASD in offspring.

Design, synthesis, and antitumor evaluation of morpholine substituted bisnaphthalimides as DNA targeting agents.

A series of mono- and bisnaphthalimides derivatives containing 3-nitro and 4-morpholine moieties were designed, synthesized, and evaluated for their in vitro anticancer activities against four cancer cell lines. Some compounds exhibited relatively good antiproliferative activity on the cell lines tested, in comparison with mitonafide and amonafide. It is noteworthy that bisnaphthalimide A6 was identified as the most potent compound in anti-proliferation against MGC-803 cells, with an IC50 lowered to 0.09 µM, a far greater potency than that of mono-naphthalimide A7, mitonafide, and amonafide. A gel electrophoresis assay revealed that DNA and Topo I were the potential targets of compounds A6 and A7. The treatment of CNE-2 cells with compounds A6 and A7 resulted in an S phase cell cycle arrest, accompanied by the upregulation of the expression levels of the antioncogene p27 and the down-regulation of the expression levels of CDK2 and cyclin E. In addition, compounds A6 and A7-induced apoptosis was further confirmed by flow cytometry, ROS generation assay, and Hoechst 33,258 staining. In particular, in vivo antitumor assay results revealed that bisnaphthalimide A6 exhibited potent anticancer efficiency in an MGC-803 xenograft tumor model, in comparison with mitonafide, and had lower toxicity than mono-naphthalimide A7. In brief, the results suggested that bisnaphthalimide derivatives containing 3-nitro and 4-morpholine moieties might serve as DNA binding agents for the development of new antitumor agents.

[SPARC Overexpression Enhances the Sensitivity of SKM-1 Cells to Ara-C by Regulating CPBP/MLKL].

OBJECTIVE: To investigate the effect of SPARC gene overexpression on the chemotherapeutic sensitivity of AML-MDS cell line SKM-1 to Ara-C and to further explore its mechanism. METHODS: Subjects were divided into 6 groups: SKM-1 cells (Control), Negative control (LV-NC), SPARC overexpression (LV-SPARC), SKM-1 cells+30 ng/ml Ara-C (30 ng/ml Ara-C), LV-NC+30 ng/ml Ara-C and LV-SPARC+30 ng/ml Ara-C. Cell activity was detected by CCK-8 assay, cell cycle distribution and apoptosis were detected by flow cytometry, mRNA expression levels of SPARC, CPBP and MLKL were detected by RT-qPCR, and the expression levels of related protein were detected by Western blot. RESULTS: After co-treatment with SPARC overexpression and Ara-C, the cell viability decreased and apoptosis increased significantly, with obvious up-regulation of Bax and down-regulation of BCL-2 (P<0.05). Compared with the control group, the cell cycle of LV-SPARC+30 ng/ml Ara-C group was significantly arrested in S phase with obvious down-regulation of CDK2 and up-regulation of p27KIP1 (P<0.05). Compared with LV-SPARC group and 30 ng/ml Ara-C group, the mRNA and protein expression levels of CPBP and MLKL (p-MLKL) were significantly elevated in LV-SPARC+30 ng/ml Ara-C group (P<0.05). In addition, after co-treatment with SPARC overexpression and Ara-C, the protein expression level of p-AKT decreased and the protein expression level of p53 increased (P<0.05). CONCLUSION: SPARC overexpression enhanced the sensitivity of SKM-1 cells to Ara-C and promoted cell cycle arrest and apoptosis, the mechanism of which may be related to the regulation of CPBP/MLKL pathway.

[Clinical efficacy of oblique lumbar interbody fusion combined with unilateral pedicle screw fixation via Wiltse approach in the treatment of lumbar spinal stenosis].

OBJECTIVE: To evaluate the efficacy of oblique lumbar interbody fusion combined with unilateral pedicle screw fixation via Wiltse approach in the treatment of lumbar spinal stenosis. METHODS: From July 2017 to January 2019, 90 patients with lumbar spinal stenosis, including 38 males and 52 females, aged from 43 to 75 years old with an average of(59.9±8.8) years old, and were treated with oblique lumbar interbody fusion(OLIF) combined with Wiltse unilateral pedicle screw fixation. Surgical decompression and fixation was performed in 50 cases of single segment, 32 cases of double segments and 8 cases of three segments. The distribution of responsible segments included 8 cases of L2-L3, 12 cases of L3-L4 and 30 cases of L4-L5 on single segment, 10 cases of L2-L4 and 22 cases of L3-L5 on double segments, and 8 cases of L2-L5 on three segments. The operation time, blood loss and occurrence of complications were recorded, Visual analogue scale(VAS), Oswestry Disability Index(ODI) and SF-36 scale were used to evaluate clinical efficacy. Lumbar X-ray and MRI were taken at three days after operation, interverterbral space height, intervertebral foraminal height, interverterbral foraminal area, and spinal canal area were measured, and interbody fusion was evaluated according to CT at half a year after operation. RESULTS: All patients were followed up from 12 to 33 months, with an average of (20.2±6.6) months. Mean operation time was (103.3±35.9) min, and mean intraoperative blood loss was (70.4±17.8) ml. VAS of low back pain leg pain, and ODI decreased from 6.2±1.1, 6.1±0.9 and (59.9±4.2)% to 2.7±0.5, 2.5±0.5 and (31.3±8.8)%. SF-36 scale significantly increased from (37.2±3.1) to (54.9±6.1) at the six months postoperation(P<0.05). The intervertebral space height, intervertebral foraminal height, intervertebral foraminal area, and spinal canal area were significantly improved at 3 days after operation(P<0.05). Six months after operation, CT scan showed well fusion in 87 cases, but 3 cases with poor fusion, including 1 case of single segment, 2 cases of multi-segments. The total fusion rate was 96.7% (87/90), the single segment fusion rate was 98.0% (49/50), and the multi-segments fusion rate was 95.0%(38/40). The overall incidence of complications was 17.8%(16/90), including transient iliopsoas muscle weakness in 5 cases (5.6%), endplate fracture in 2 cases (2.2%), peritoneal injury in 1 case (1.1%), postoperative hematoma in 1 case (1.1%), adjacent segment disease in 1 case(1.1%), and fusion cage subsidence in 6 cases (6.7%). Three patients was followed up for recurrent nerve root pain and the symptoms were relieved after revision operation. All complications were relieved or disappeared in varying degrees during the follow-up period, and there were no complications such as cage displacement and screw fracture. CONCLUSION: OLIF combined with unilateral pedicle screw fixation via Wiltse approach is effective in treating lumbar spinal stenosis with minimally invasive advantages such as less trauma and less complications. Under the premise of strictly grasping the indications, this method could also achieve satisfactory clinical results in multi-segments oprations.

Distinctive Impact of Heat Treatment on the Mechanical Behavior of Nacreous and Crossed-Lamellar Structures in Biological Shells: Critical Role of Organic Matrix.

As biological ceramic composites, mollusk shells exhibit an excellent strength-toughness combination that should be dependent on aragonite/organic matrix interfaces. The mechanical properties and fracture mechanisms of the nacreous structure in the Cristaria plicata (C. plicata) shell and crossed-lamellar structures in the Cymbiola nobilis (C. nobilis) shell were investigated, focusing on the critical role of the organic matrix/aragonite interface bonding that can be adjusted by heat treatments. It is found that heat treatments have a negative impact on the fracture behavior of the nacreous structure in the C. plicata shell, and both the bending and shear properties decrease with increasing heat-treatment temperature because of the loss of water and organic matrix. In contrast, for the crossed-lamellar structure in C. nobilis shell, the water loss in heat treatment slightly decreases its bending properties. When the organic matrix is melted after an appropriate heat treatment at 300°C for 15 min, its bending properties can be greatly enhanced; in this case, remarkable toughening mechanisms involving crack deflection and the fiber pull-out are exhibited, although the interfacial bonding strength reduces. Therefore, an appropriate heat treatment would bring about a positive impact on the fracture behavior of crossed-lamellar structure in the C. nobilis shell. The major research findings have provided an important inspiration that the inducement of moderately weak interfaces rather than all strong interfaces might enhance the comprehensive mechanical properties of fiber-reinforced ceramic composites.

An Ingenious Microstructure Arrangement in Deep-Sea Nautilus Shell against the Harsh Environment.

Mollusk shells generally consist of several macro-layers with different microstructures. To explore the specific role that different macro-layers play in the overall mechanical properties of shells, the microstructures, hardness distribution, and three-point bending behavior in the deep-sea Nautilus shell were investigated. It is found that the shell presents a hierarchical structure comprising three layers in thickness, that is, the outer, middle, and inner layers, which exhibit homogeneous, prismatic, and nacreous structures, respectively. Among them, the homogeneous structure in the outer layer is harder, which is beneficial for the shell to enhance resistance to wear and perforation. Furthermore, both the bending strength and fracture energy for group Up (loading from outer to inner surfaces) are far higher than those for group Down (loading from inner to outer surfaces), indicating that the inner nacreous layer is not only stronger but also tougher. Cracks tend to deflect at the interfaces in nacreous structure, and nacreous structure is thereby more resistant to breakage. Hence, the nacreous structure in the inner layer could protect the shell from breaking catastrophically in the deep sea with high pressure. In brief, the combination of a harder outside layer and a tougher inside layer provides an effective protective structure for the deep-sea shell, and the excellent environment adaptability of Nautilus shell can thus be interpreted in terms of its ingenious microstructure arrangement.

Treatment of Degenerative Lumbar Scoliosis with Oblique Lumbar Interbody Fusion in Conjunction with Unilateral Pedicle Screw Fixation via the Wiltse Approach.

OBJECTIVE: To evaluate the clinical outcomes of oblique lumbar interbody fusion (OLIF) in conjunction with unilateral pedicle screw fixation (UPSF) via the Wiltse approach in treating degenerative lumbar scoliosis (DLS). METHODS: The article is a retrospective analysis. Twelve patients with DLS who underwent combined OLIF and UPSF between July 2017 and December 2018 were included. The study included 2 male and 10 female patients, with a mean age at the time of the operation of 67.2 ± 9.1 years. The surgical characteristics and complications were evaluated. The clinical and radiological data such as the correction of deformity, coronal and sagittal profile were analyzed. RESULTS: The mean follow-up time of the study was 26.8 ± 1.8 months. At the final follow-up, all patients who underwent combined OLIF and UPSF achieved statistically significant improvements in coronal Cobb angle (from 19.6° ± 4.8° to 6.9° ± 3.8°, P < 0.01), distance between the C7 plumb line and central sacral vertebral line (from 2.5 ± 1.7 cm to 0.9 ± 0.6 cm, P < 0.01), sagittal vertebral axis (from 4.3 ± 4.3 cm to 1.5 ± 1.0 cm, P = 0.03), lumbar lordosis (from 29.4° ± 8.6° to 40.8° ± 5.8°, P < 0.01), pelvic tilt (from 27.6° ± 10.8° to 18.3° ± 7.0°, P < 0.01), pelvic incidence-lumbar lordosis mismatch (from 23.3° ± 10.5° to 11.9° ± 8.4°, P < 0.01), and cross-sectional area of the dural sac (from 87.33 ± 39.41 mm2 to 124.70 ± 39.26 mm2 , P < 0.01). The visual analogue score for back and leg pain and Oswestry Disability Index of all patients significantly improved postoperatively (P < 0.01). One case of lumbar plexus injury was found after surgery. During the follow-up period, one patient had cage subsidence. A fusion rate of 100% and good positioning of the pedicle screws were achieved in all patients at the final follow-up. CONCLUSION: OLIF in conjunction with UPSF is a safe and effective minimally invasive procedure for correcting both coronal and sagittal deformities, as it results in an improved quality of life in patients with DLS.

Measurement consistency of dynamic stretching muscle stiffness evaluated using shear wave elastography: comparison among different stretched levels and ROI sizes.

AIM: To investigate the reliability of quantitative analysis of dynamic stretching muscle stiffness using shear wave elas-tography (SWE), and to evaluate the influence of stretched levels and region of interest (ROI) sizes on the repeatability of SWE measurements. MATERIALS AND METHODS: SWE videos of the gastrocnemius medius were collected during ankle movement from plantar flexion (PF) 40° to dorsiflexion (DF) 30°. Shear wave imageswere collected of ankle angles at PF 25°, 0°, DF 15°, and DF 30°, representing the slack status, mildly stretched level, moderately stretched level, and maximal stretched level of the gastrocnemius medius, respectively. ROI circles with diameters of 2 mm, 5 mm, and 8 mm were applied to measure the shear modulus. Intra-observer, and inter-observer repeatability of the measurements were compared among different stretched levels and ROI sizes. RESULTS: Twenty-one healthy volunteers were enrolled. Muscle stiffness increased as the ankle DF increased. Intraclass correlation coefficients (ICCs) of intra-observer and inter-observer repeatability obtained for ROI sizes of 2 mm, 5 mm and 8 mm indicated good to excellent repeatability at all stretched levels. CONCLUSIONS: Shear wave elastography appeared to be a reliable tool to evaluate the dynamic stretching muscle stiffness with satisfactory repeatability at various stretched levels of gastrocnemius medius. Good to excellent repeatability was found using different ROI sizes.

Quantitative evaluation of passive muscle stiffness by shear wave elastography in healthy individuals of different ages.

OBJECTIVES: To investigate age-related changes on passive muscle stiffness in healthy individuals and measure the shear modulus in different age groups. METHODS: Shear wave elastography (SWE) movies of gastrocnemius medialis (GM) were collected during passive stretching induced by ankle rotation from plantarflexion (PF) to dorsiflexion (DF). A series of SWE images at ankle angles of PF 40°, PF 30°, PF 20°, PF 10°, 0°, DF 10°, DF 20°, and DF 30° were collected and shear moduli measured accordingly for analyses. RESULTS: Eighty-six healthy volunteers (27 children, 31 middle-aged adults, and 28 older people) were recruited. No significant difference was observed in the shear modulus between the three groups at ankle angles of PF 40°, PF 30°, PF 20°, PF 10°, and 0° (p > 0.05). The difference in the shear modulus among the three groups became significant as DF increased. At ankle angles of DF 10°, DF 20°, and DF 30°, the shear modulus was the greatest in the older group, followed by the middle-aged group and then the children group (p = 0.007, 0.000, and 0.000, respectively). CONCLUSIONS: Passive muscle stiffness increases with age, and the difference between age groups was pronounced only after reaching a certain degree of stretching. KEY POINTS: • The influence of age on passive muscle stiffness becomes pronounced only after reaching a certain degree of stretching. • Age should be considered when evaluating passive muscle stiffness in muscular disorders.

[Inhibitory Effects of Metformin on Proliferation of SKM-1 Cells and The Related Underlying Mechanisms].

OBJECTIVE: To investigate the effects of metformin on the proliferation of AML-MDS cells (SKM-1 cells) and its related mechanisms. METHODS: CCK-8 was used to test the cell proliferation; Flow cytometry was used to detect the cell apoptosis and cell cycle; Western blot was used to test the expression level of AMPK and cell cycle regulatory proteins. RESULTS: Metformin could inhibit the proliferation of SKM-1 cells, which may be attributed to metformin-induced cell cycle arrest in G0/G1 but not to metformin induced cell apoptosis. The expression levels of G1-related protein CyclinD1 and CDK4 were down-regulated, while the expression levels of P53, P21CIP1 and P27kIP1 were up-regulated. Moreover, the phosphorylation level of AMPK was up-regulated. CONCLUSION: Metformin inhibits the proliferation of SKM-1 cells, which may relate with AMPK-induced cell cycle arrest. However, future studies are necessary to further explore the related mechanisms.

[Changes in Metabolism and Function of Platelets Stored at Different Temperature].

AAbstractObjective:To compare and analyze the metabolic and functional changes in platelets stored at 4 ℃ and ones stored at 22 ℃ with agitation so as to provide an experimental basis for the cryopreservation technology of platelets. METHODS: Samples were collected from platelets stored at 4 ℃ in 2, 4, 6, 11, 15 and 21 days, and from ones stored at 22 ℃ with agitation during the same days, the metabolism indicators and thromboelastogram (TEM) were analysed. RESULTS: In metabolism, there were no significant changes of pH, GLU，PCO2, PCO2 and MPV levels of platelets stored at 4 ℃ for <6 days (P＞0.05), However, the Plt count decreased, the PDW and LDH level incrseased (P＜0.05). At the same time, only MPV had no changes of platelets stored at 22 ℃ during above-mentioned same days (P＞0.05), while the pH, PCO2, GLU, Plt all decreased, and PO2, LDH, PDW incrseased (P＜0.05). There were significant changes about the pH value, PO2, Plt, MPV, LDH, GLU levels between the two kinds of stored platelets during the same storing period (P＜0.05). The pH value and MPV of platelets stored at 4 ℃ were obviously lower than ones stored at 22 ℃, while GLU, PO2, LDH and Plt levels showed reverse changes (P＜0.05). Meanwhile, the PCO2 of platelets stored at 4 ℃ not could be detected and the Plt count reduced rapidly from d15. In function, the MA level of platelets stored at 4 ℃ was slower than that of platelets stored at 22 ℃, that is, the MA level of platelets stored at 4 ℃ were higher than that of platelets stored at 22 ℃ during the same storeing period (P＜0.05). CONCLUSION: Platelets stored at 4 ℃ have much slower metabolism than ones stored at 22 ℃, and the aggregation is stronger of platelets stored at 4 ℃ than that of ones at 22 ℃ during the same conservation period.

Carbapenem-resistant Klebsiella pneumoniae infection causing rupture of graft artery in solid organ recipients: Case report and review of literature.

RATIONALE: Donor-derived bacterial infection is a rare cause of morbidity after solid organ transplantation (SOT) but associated with significant morbidity and mortality, deaths caused by carbapenem-resistant Klebsiella pneumoniae (CRKP) infection account for a considerable proportion of postoperation mortality rate in liver and kidney recipients. The arterial rupture as a result of fungal arteritis is occasionally described, while the rupture of graft vascular anastomosis after SOT due to donor-derived CRKP infection is rarely reported. PATIENTS CONCERNS: We reported 1 patient with donor-derived CRKP infection following liver transplantation and 2 patients following renal transplantation (1 liver and 2 kidneys were from the same donor), who experienced sudden abdominal pain and abdominal hemorrhage almost at the same time after organ transplantation. DIAGNOSIS: The patients were diagnosed as graft arteries rupture due to corrosion caused by CRKP infection based on computed tomography scan, blood culture, laparotomy, and pulse-field gel electrophoresis. INTERVENTIONS: Anti-shock treatment, exploratory laparotomy, broad-spectrum antibiotics, and abdominal puncture and drainage were given. OUTCOMES: The liver recipient survived as well as the liver graft, still under treatment of multiple abdominal infections. The 2 renal recipients were alive after resection of the renal grafts and underwent hemodialysis. LESSONS: Rupture of graft artery should be foreseen when donor-derived CRKP infection was confirmed and broad-spectrum antibiotics and other interventions need to be considered.

Characterization of acute renal allograft rejection by proteomic analysis of renal tissue in rat.

Rapid and reliable biomarkers of renal allograft rejection have not been available. This study aimed to investigate biomarkers in renal allograft tissue using proteomic analysis. Orthotopic kidney transplantations were performed using Fisher (F344) or Lewis rats as donors and Lewis rats as recipients. Syngenic control group (Group I) constituted F344-to-F344 orthotopic kidney allo-transplantations (n = 8); and allogenic group (Group II) consisted of F344-to-Lewis orthotopic kidney allo-transplantations (n = 8). Renal tissues were harvested 7 days after transplantation. Samples were analyzed using 2-D electrophoresis and matrix assisted laser desorption ionization-time of flight mass spectrometry. 6 differentially expressed proteins were identified between allogenic group and syngenic control group. A rat model of acute renal allograft rejection was successfully set up. Differentially expressed proteins in renal allograft tissue of rat were detected using proteomic analysis and might serve as novel diagnostic and therapeutic targets in human. Quantitative proteomics, using MALDL-TOF-MS methodology has the potential to provide a profiling and a deeper understanding of acute renal rejection.