TBC1D15 deficiency protects against doxorubicin cardiotoxicity via inhibiting DNA-PKcs cytosolic retention and DNA damage

Clinical application of doxorubicin (DOX) is heavily hindered by DOX cardiotoxicity. Several theories were postulated for DOX cardiotoxicity including DNA damage and DNA damage response (DDR), although the mechanism(s) involved remains to be elucidated. This study evaluated the potential role of TBC domain family member 15 (TBC1D15) in DOX cardiotoxicity. Tamoxifen-induced cardiac-specific Tbc1d15 knockout (Tbc1d15CKO) or Tbc1d15 knockin (Tbc1d15CKI) male mice were challenged with a single dose of DOX prior to cardiac assessment 1 week or 4 weeks following DOX challenge. Adenoviruses encoding TBC1D15 or containing shRNA targeting Tbc1d15 were used for Tbc1d15 overexpression or knockdown in isolated primary mouse cardiomyocytes. Our results revealed that DOX evoked upregulation of TBC1D15 with compromised myocardial function and overt mortality, the effects of which were ameliorated and accentuated by Tbc1d15 deletion and Tbc1d15 overexpression, respectively. DOX overtly evoked apoptotic cell death, the effect of which was alleviated and exacerbated by Tbc1d15 knockout and overexpression, respectively. Meanwhile, DOX provoked mitochondrial membrane potential collapse, oxidative stress and DNA damage, the effects of which were mitigated and exacerbated by Tbc1d15 knockdown and overexpression, respectively. Further scrutiny revealed that TBC1D15 fostered cytosolic accumulation of the cardinal DDR element DNA-dependent protein kinase catalytic subunit (DNA-PKcs). Liquid chromatography-tandem mass spectrometry and co-immunoprecipitation denoted an interaction between TBC1D15 and DNA-PKcs at the segment 594-624 of TBC1D15. Moreover, overexpression of TBC1D15 mutant (∆594-624, deletion of segment 594-624) failed to elicit accentuation of DOX-induced cytosolic retention of DNA-PKcs, DNA damage and cardiomyocyte apoptosis by TBC1D15 wild type. However, Tbc1d15 deletion ameliorated DOX-induced cardiomyocyte contractile anomalies, apoptosis, mitochondrial anomalies, DNA damage and cytosolic DNA-PKcs accumulation, which were canceled off by DNA-PKcs inhibition or ATM activation. Taken together, our findings denoted a pivotal role for TBC1D15 in DOX-induced DNA damage, mitochondrial injury, and apoptosis possibly through binding with DNA-PKcs and thus gate-keeping its cytosolic retention, a route to accentuation of cardiac contractile dysfunction in DOX-induced cardiotoxicity.

Endoplasmic Reticulum Stress-Mediated p62 Downregulation Inhibits Apoptosis via c-Jun Upregulation

Cereblon (CRBN), a substrate receptor of cullin 4-RING E3 ligase (CRL4) regulates the ubiquitination and degradation of c-Jun, mediating the lipopolysaccharide-induced cellular response. However, the upstream signaling pathway that regulates this process is unknown. In this study, we describe how endoplasmic reticulum (ER) stress reversely regulates sequestosome-1 (p62)and c-Jun protein levels. Furthermore, our study reveals that expression of p62 attenuates c-Jun protein levels through the ubiquitinproteasome system. Conversely, siRNA knockdown of p62 elevates c-Jun protein levels. Immunoprecipitation and immunoblotting experiments demonstrate that p62 interacts with c-Jun and CRBN to form a ternary protein complex. Moreover, we find that CRBN knockdown completely abolishes the inhibitory effect of p62 on c-Jun. Using brefeldin A as an inducer of ER stress, we demonstrate that the p62/c-Jun axis participates in the regulation of ER stress-induced apoptosis, and that CRBN is required for this regulation. In summary, we have identified an upstream signaling pathway, which regulates p62-mediated c-Jun degradation. Our findings elucidate the underlying molecular mechanism by which p62/c-Jun axis regulates the ER stress-induced apoptosis, and provide a new molecular connection between ER stress and apoptosis.

Monitoring changes of knee in amateur marathon athletes using synthetic MRI: a preliminary study / 中华放射学杂志

Objective:To explore the value of synthetic MRI in quantitative monitoring of knee joint structural and cartilage changes of amateur marathon runners before and after the whole marathon.Methods:Totally 26 amateur marathon enthusiasts from Zhuhai City, Guangdong Province were recruited from October 2019 to January 2020. The right knee joints were scanned 1 week before the race and within 48 h after the race. The scanning sequence included the three-dimensional proton density weighted image with isotropic (3D-CUBE-PD) sequence and synthetic MRI sequence. The conventional contrast weighted images T 1WI, T 2WI, proton density (PD) weighted imaging, short-T 1 inversion recovery (STIR) and T 1, T 2, PD mapping were obtained by the latter scans. The 3D-CUBE-PD sequence was used as a reference to evaluate the detection of knee joint lesions. The knee articular cartilage was divided into 8 subregions: central medial femoral condyle (CMFC), posterior medial femoral condyle (PMFC), central lateral femoral condyle (CLFC), posterior lateral femoral condyle (PLFC), medial tibia plateau (MTP), lateral tibia plateau (LTP), patella and trochlear. Based on the synthetic MRI quantitative mapping, the T 1, T 2 and PD values of each cartilage subregion were measured independently by 2 radiologists. The ICC was used to evaluate the consistency of the measurement between observers. The T 1, T 2 and PD values of knee cartilage before and after marathon exercise were compared by Wilcoxon signed rank test. Results:The 2 radiologists had good consistency in the measurement of T 1, T 2 and PD values of knee articular cartilage with the ICC values of 0.912, 0.933 and 0.954, respectively. The synthetic MRI quantitative mapping sequence can detect all cartilage damage ( n=3) and joint effusion ( n=15), and 7 of 9 meniscus injuries were detected. The T 1, T 2 and PD values of the knee cartilage as a whole before the race were higher than those after race, and the differences were statistically significant (all P<0.05). The T 1 values were statistically significant except patellar cartilage and trochlear cartilage, and T 2 values were significantly different in the CMFC, LTP, MTP ( P<0.05). Conclusion:Synthetic MRI has a good display of knee joint structural lesions, and its quantitative parameters T 1, T 2 and PD can detect the changes of knee cartilage before and after marathon.

Efficacy and pharmacoeconomical analysis of three platinum based chemotherapy regimens for advanced non small cell lung cancer / 中国基层医药

Objective To explore the cost-effectiveness and clinical effect of three platinum based chemotherapy regiments for advanced non small cell lung cancer (NSCLC).Methods 100 patients who were diagnosed as NSCLC,were randomly divided into four groups.The group Ⅰ received NP which was given NVB and DDP.The group Ⅱreceived GP which was given GEM and DDP.The group Ⅲ received TP which was given taxotere and DDP.The clinical effect,adverse reaction and cost effectiveness of the three groups were assessed.Results The clinical effective rates of the three groups were 31.43%,36.36%,37.50% from Ⅰ to Ⅲ group.The adverse events of the group Ⅰ and group Ⅱ were more than those of the group Ⅲ.In the adverse effects of treatment,the major cytotoxicity was digestive reaction and leukopenia in the two groups,but they were tolerable.The ratios of cost effectiveness in the four groups were 550.22yuan,556.48yuan,583.23yuan from Ⅰ to Ⅲ group.Conclusion The NP group is the best one in total cost.

Effect of Autophagy on Process of Phosphate Induced Vascular Smooth Muscle Cell Calcification in Experimental Rats / 中国循环杂志

Objective: To explore the effect of autophagy on process of high phosphate salt induced vascular smooth muscle cell (VSMC) calciifcation in experimental rats. Methods: Rats’ model of VSMC calciifcation was induced by phosphate incubation. VSMC were divided into 3 groups:①Control group,②Calciifcation group which included 3 subgroups as 4-day subgroup, the cells were cultured by 3.2 mmol/L phosphate for 4 days, 6-day subgroup and 8-day subgroup,③Calciifcation+ 3-MA (autophagy inhibitor) group, in which the 8-day cells were cultured with 5mmol/L 3-MA. Calcium nodule formation and calcium deposition in VSMC were measured by Alizarin red staining and o-cresolphthaleincomplexone method, protein expressions of Runx2, α-SMA and LC3 II were examined by Western blot analysis, autophagosome formation in VSMC was measured by transmission electron microscope and the localization and expression of Runx2 and LC3 II in VSMC were observed by immunolfuorescent microscope. Results: Compared with Control group, the cells at 8-day subgroup showed more calcium nodules, higher calcium deposition, increased protein expressions of Runx2, LC3 II, more autophagosome and decreased α-SMA expression, allP<0.05. Compared with 8-day subgroup, the cells in Calcification+3-MA group presented increased calcium deposition, decreased lfuorescence distribution of LC3 II and more cells with positive Runx2 protein expression, all P<0.05. Conclusion: Autophagy has the protective effect on process of phosphate induced VSMC calciifcation in experimental rats.

Clinical research of chronic pelvic cavity pain syndrome treated with acupoint catgut embedding therapy / 中国针灸

<p><b>OBJECTIVE</b>To explore the clinical efficacy of acupoint catgut embedding therapy on chronic pelvic cavity pain syndrome differentiated as kidney deficiency and stagnation of damp heat, and explore the impacts on plasma P substance (SP), plasma beta-endorphin (β-EP).</p><p><b>METHODS</b>One hundred and eighty cases were randomly divided into a catgut embedding group (90 cases) and a western medication group (90 cases). In the western medication group, tamsulosin capsules 0. 2 mg were prescribed for oral administration, once a day; indometacin sustained release tablets, 25 mg, three times a day. Totally, the oral administration for 8 weeks was required. In the catgut embedding group, the acupoint catgut embedding therapy was applied to Qugu (CV 2), Shenshu (BL 23), Zhibian (BL 54), Huiyin (CV 1) and Sanyinjiao (SP 6), once every two weeks; the treatment of 4 weeks made one session, and two sessions were required. Before and after treatment, TCM symptom score, NIH-CPSI (the National Institute of Health Chronic Prostatitis Symptom Index) score, lecithin body numbers in prostatic fluid, score in SAS (self-rating anxiety scale), score in SDS (self-rating depression scale), the levels of SP and β-EP, etc. were observed in the two groups, and the clinical efficacy was assessed in the two groups.</p><p><b>RESULTS</b>(1) Ten cases were dropped in either group. The total effective rate was 91. 25% (73/80) in the catgut embedding group, higher than 78. 75% (63/80) in the western medication group (P<0. 05). (2) After treatment, TCM symptom score, total score in NIH-CPSI, pain score and the scores in SAS and SDS were all reduced as compared with those before treatment in the two groups (all P<0. 05). After treatment, TCM symptom score, total score and pain score in NIH-CPSI, and the scores in SAS and SDS in the catgut embedding group were both lower than those in the western medication group (all P<0. 05). (3) After treatment, the lecithin body numbers were both increased as compared with those before treatment in the two groups (both P<0. 05), and the result in the catgut embedding group was higher than that in the western medication group (P<0. 05). (4)After treatment, the SP level was lower than that before treatment in the two groups (both P<0. 05); the level of p-EP was increased as compared with that before treatment (both P<0. 05). The SP level in the catgut embedding group was lower than that in the western medication group (P<0. 05); the level of β-EP was higher than that in the western medication group (P<0. 05).</p><p><b>CONCLUSION</b>The acupoint catgut embedding therapy apparently relieves the clinical symptoms of chronic pelvic cavity pain syndrome differentiated as kidney deficiency and stagnation of damp heat as well as the condition of anxiety and depression, increases lecithin body numbers in prostatic fluid and β-EP level and reduces SP level.</p>

Modified anterior approach for treatment of ulnar coronoid process fracture / 中华创伤杂志

Objective To investigate the clinical effect of modified anterior approach to manage fracture of the ulnar coronoid process via the space of brachial artery and vein with median nerve.Methods From June 2012 to January 2013,11 patients with ulnar coronoid fracture were fixed via the modified anterior approach.The operation time,intraoperative blood loss and postoperative complications were recorded.Flexion and rotation range of motion about the injured and normal elbow were observed during postoperative follow-up period.Function of elbow joint was evaluated by mayo elbow performance index (MEPI).Results There was approximate 8 cm in length and 5 cm in width between the brachial vessels and median nerve.Operated angle from radial to ulnar side was fifty degrees and from proximal to distal end was sixty degrees.All the patients were available for follow-up.The fracture healed,that is the elbow flexion restored [(130.7 ±5.0) °] was 96.6％ of the unaffected elbow,elbow extension restored [(7.6 ± 8.1) °] was 84.0％ of the unaffected elbow,pronation restored [(86.9 ± 3.8) °] was 98.2％ of the unaffected side,and supination restored [(85.6 ± 6.0) °] was 96.7％ of the unaffected side.MEPI of the elbow joint was over 75 points.Conclusion Modified anterior approach is relatively safe and simple in operation and results in satisfactory function recovery of the elbow joint,providing a new surgical approach for treatment of coronoid process fracture.

Clinical observation of docetaxel combined with carboplatin as second -or third -line treatment for patients ;with advanced triple -negative breast cancer / 中国基层医药

Objective To observe the clinical efficacy and adverse reactions of docetaxel and carboplatin 21 day regimen (TC)as second -or third -line in treatment of patients with advanced triple -negative breast cancer (ATNBC).Methods 37 ATNBC patients with anthracyclines resistance were given docetaxel 70mg/m2 ,the first day of intravenous drip,carboplatin AUC =5,through intravenous drip on the first day,21 days for a course of treatment, until disease progression or not tolerated or most received six cycles of chemotherapy.Results 147 cycles of chemo-therapy were completed in 37 patients,and the median chemotherapy cycle was 4 cycles.The efficacy and adverse reactions could be evaluated in all patients.Among them,complete remission (CR)in 0 patient,partial remission (PR)in 12 patients (32.43%),stable (SD)in 11 patients (29.73%),and progression (PD)in 14 patients (41.18%).The effective rate(CR +PR)was 32.43%.The disease control rate (CR +PR +SD)was 62.15%,median TTP was 3.9 months (95%CI:4 -8months),and median OS was 10 months.Until the end of follow -up,there were 3 patients with no progress.The main adverse reactions were hematological toxicity (51.53%)and gastrointestinal reaction (56.76%).Conclusion TC regimen is effective and safe in the treatment of patients with ATNBC.

Effect of Polypeptide Extract from Scorpion Venom on MMP2 and MMP9 Expression in Leukemia -NOD/SCID Mice / 天津医药

Objective: To investigate the effect of polypeptide extract from scorpion venom (PESV) on the matrix metalloproteinase2（MMP2）and matrix metalloproteinase9（MMP9）expression in leukemia-NOD/SCID mice, and the intervention mechanism of PESV in the multiplication and infiltration of leukemic cells thereof. Methods: In order to establish the animal model of outer marrow infiltration of human leukemia，bone marrow mononuclear cells of leukemia patients, irradiated 270 cGy on body by ~(137)Cs, were injected into NOD/SCID mice. The mice were randomly divided into five groups. The groups I, II and III were treated with different concentrations of PESV. Group Ⅳ was the model group injected by the normal saline solution. GroupⅤ was taken as control. The peripheral white blood cell count and blood smear were observed in groups. All of the mice were killed after four-week observation and MMP2 and MMP9 expressions were examined using Real timePCRmethod. Results: The expression levels of MMP2 and MMP9 were significantly lower in group I, group II and group III than that of the model group(P < 0.05). The expression levels of MMP2 and MMP9 were related to the concentration of PESV. Moreover, the peripheral white blood cell count and blood smear were more normal in mice treated with PESV than those of mice of model group. Conclusion: PESV inhibited the overexpression of MMP2 and MMP9 in leukemia-NOD/SCID mice, which significantly inhibited the multiplication and infiltration of leukemic cells.

Study on 3-nminobenzamide enhancing the radiosensitivity of human esophageal carcinoma cell strain in vitro / 中华放射医学与防护杂志

Objective To evaluate the ability of 3-AB to sensitize the human esophageal carcinoma cell strain (CaEs-17) to radiation in v/tro and its mechanisms. Methods CaEs-17 cells were treated with 3-AB at 0, 2.5, 7.5 mmol/L and given irradiation O, 3, 6, 9, 12 Gy. 3-AB concentration in each group was made dose-survival curve using multi-target single-hit maiths model by clonogenie assay. MTT assay was performed to observe the survival of irradiated cells.comet assay and metaphase chromosome analysis were used to measure the DNA damage degree and chromosome aberration of CaEs-17 cell after 3-AB treatment and irradiation. Results Cell survival experiments showed SER of 1.21, 1.52 for 2.5 mmol/L, 7.5 mmol/L 3-AB respectively using multi-target single-hit maths model. The survival fraction of irradiated CaEs-17 cell was decreased after 3-AB treatment. DNA damage and the chromatid breakage number of irradiated CaEs-17 cells were increased after 3-AB treatment. Conclusions 3-AB, a PARP inhibitor, can enhance the radiosensitivity of human esophageal carcinoma cell strain (CaEs-17). DNA damage repair inhibition by 3-AB might be one of the mechanisms.

Hemodynamics of deep vein in lower limb during perioperative period of total hip arthroplasty / 中华创伤杂志

Objective To investigate hemodynamic changes of deep vein in lower limb during the perioperative period of total hip arthroplasty (THA) and early diagnosis of deep vein thrombosis (DVT). Methods Doppler ultrasound, hemorheology detection and plasma D-dimer testing were done on 62 patients treated with THA. Statistical analysis was carried out on the data of patients with or without DVT to study the early diagnosis of DVT. Results The results of Doppler ultrasound showed DVT in 8 patients. Compared with postoperative concentration of plasma D-dimer, the preoperative con-centration of plasma D-dimer was significandy higher in patients with or without DVT (P < 0.05). The levels of hemorheological indices were significantly increased at postoperative day 7 (P < 0.05). Con-clusions Doppler ultrasound combined with plasma D-dimer testing and hemorheology detection are helpful in early diagnosis of DVT.