Minocycline inhibits neuroinflammation and enhances vascular endothelial growth factor expression in a cerebral ischemia/reperfusion rat model

BACKGROUND： Brain ischemia involves secondary inflammation, which significantly contributes to the outcome of ischemic insults. Vascular endothelial growth factor （VEGF） may play an important role in the vascular response to cerebral ischemia, because ischemia stimulates VEGF expression in the brain, and VEGF promotes formation of new cerebral blood vessels. Minocycline, a tetracycline derivative, protects against cerebral ischemia and reduces inflammation, oxidative stress, and apoptosis. OBJECTIVE： To observe the influence of minocycline on VEGE interleukin-1 beta （IL-119 ）, and tumor necrosis factor alpha （TNF-α） expression in Wistar rats with focal cerebral ischemia/reperfusion injury, and to study the neuroprotection mechanism of minocycline against focal cerebral ischemia/reperfusion injury. DESIGN, TIME AND SETTING： Randomized, controlled experiment, which was performed in the Chongqing Key Laboratory of Neurology between March 2007 and March 2008. MATERIALS： A total of 36 female, Wistar rats underwent surgery to insert a thread into the left middle cerebral artery. Animals were randomly divided into sham-operation, minocycline treatment, and ischemia/reperfusion groups, with 12 rats in each group. Minocycline （Huishi Pharmaceutical Limited Company, China） was dissolved to 0.5 g/L in normal saline. METHODS： A 0.5-1.0 cm thread was inserted into rats from the sham-operation group. Rats in the ischemia/reperfusion group underwent ischemia and reperfusion. The minocycline group received minocycline （50 mg/kg） 12 and 24 hours following ischemia and reperfusion, whereas the other groups received saline at the corresponding time points. MAIN OUTCOME MEASURES： mRNA and protein expression of IL-1β and TNF-α was measured by reverse transcriptase-polymerase chain reaction （RT-PCR） and enzyme linked immnnosorbent assay （ELISA）, respectively. VEGF mRNA and protein expression was examined by RT-PCR, Western blot, and ELISA. RESULTS： Minocycline decreased the focal infarct volume. VEGF, IL-1β, and TNF-α expression was upregulated in the ischemia-perfusion group after injury. Following minocycline treatment, IL-1β and TNF- α expression was significantly downregulated, and VEGF was significantly upregulated, compared with the ischemia/reperfusion group （all P 〈 0.01）. Expression of VEGF, IL-1β, and TNF- α was greater in the ischemia-perfusion and minocycline treatment groups, compared with sham-operated animals （P 〈 0.01 ）. CONCLUSION： Minocycline can reduce expression of IL- 1β and TNF-α, and increase VEGF expression, in the rat brain following cerebral ischemia/reperfusion. From these findings, a hypothesis can be formed that minocycline attenuates neuroinflammation and enhances recovery of vascular integrity during the process of cerebral ischemia/reperfusion.

Riboflavin/ultraviolet A-induced collagen cross-linking in rabbit corneal scar

AIM: To evaluate the biomechanical stability of the corneal scar treating with riboflavin and ultraviolet A(UVA). METHODS: Totally 86 New Zeal rabbits were divided into control group(group A, n=8) and trauma groups [group B(n=27), group C(n=24) and group D(n=27)]. Then groups B, C and D were divided into three sub-groups according to the time points of sacrifice, i.e. groups Ba, Ca and Da(4 wk, n=8); Bb, Cb and Db(6 wk, n=8); Bc(n=11), Cc(n=8) and Dc(8 wk, n=11). The right corneas of these 78 rabbits in the trauma groups were penetrated. Group B were only sutured. Group C were treated with corneal cross-linking(CXL) immediately after suturing. Group D were treated with CXL seven days after suturing. The corneal scar strips of 4.0×10.0 mm2 were cut and the stress and Young's modulus at 10% strain were evaluated. Samples from the three rabbits of group Bc and three of group Dc were used to measure the expression of alpha smooth muscle action(α-SMA). RESULTS: The mechanical strength of the corneal scar increased with time, and was strongest at 8 wk after the injury. The ultimate stress of corneal scar(group D) were 2.17±0.52 MPa, 2.92±0.63 MPa, and 4.21±0.68 Mpa at 4 wk, 6 wk and 8 wk, respectively; Young's modulus were 10.94±1.57 MPa, 11.16±2.50 MPa, and 13.36±2.10 Mpa, which were higher than that of other groups except for normal control. The expression of α-SMA in group B and group D were 0.28±0.11 and 0.65±0.20, respectively, and the difference was statistically significant(P=0.048). CONCLUSION: CXL with riboflavin/UVA at seven days after suturing improved the biomechanical properties of corneal scars most effectively in the present study.

Growth Inhibiton of Human Breast Cancer Cell Line MDA-MB-231 by Rosiglitazone through Activation of PPARγ

OBJECTIVE To investigate the anti-proliferative effect of rosiglitazone and its relationship to peroxisome proliferator-activated receptor γ （PPARγ） in human breast cancer cell line MDA-MB-231 and evaluate the potential application value of rosiglitazone for breast cancer therapy. METHODS The cytostatic effect of rosiglitazone on MDA- MB-231 cells was measured by the MTT assay. Cell-cycle kinetics was assessed by flow cytometry. Apoptotic cells were determined by the TUNEL assay. MDA-MB-231 cells were treated with rosiglitazone or in combination with the PPARy antagonist GW9662 to investigate the effect of rosiglitazone on cell proliferation and its relationship to PPARγ. RESULTS The results showed that rosiglitazone could inhibit growth of MDA-MB-231 cells in a dose- and time-dependent manner with an IC50 value of 5.2 μmol/L at 24 h after the drug was added into the culture. Cell cycle analysis showed that the percentage of G0/G1 phase cells increased, S phase cells decreased, and cells were arrested in G1 phase with increasing concentrations of rosiglitazone. Detectable signs of apoptotic cell death caused by rosiglitazone occurred at a concentration of 100 μmol/L and the apoptotic rate was （18 ± 3）%. PPARγ selective antagonist GW9662 could partially reverse the inhibitory effect of rosiglitazone on proliferation of MDA-MB-231 cells. CONCLUSION It was concluded that rosiglitazone can inhibit growth of MDA-MB-231 cells via PPARy activation and a high concentration of rosiglitazone can also induce MDA-MB-231 cell apoptosis. These results suggest that PPARy represents a putative molecular target for chemopreventive therapy and rosiglitazone may be effective in the treatment of breast cancer.

MG132 Inhibits Myocardial Ischemia-reperfusion Injury by Regulating Apoptotic Pathway

Objectives To administrated proteasome inhibitor-MG-132 prior to reperfusion in rat myocardial ischemia-reperfusion model to determine whether MG-132 could reduce myocytic apoptosis. Methods and results MG-132 （0. 75 mg/kg in 2 ml DMSO） injection 5 min prior to reperfusion resulted significant reduction of myocardial reperfusion injury. This effect was accompanied by reduced polymorphonuclear neutrophils （PMN） infiltration in myocardial region surrounding the myocardial infarct, reduced apoptosis in cardiac myocytes, reduced NF-κB activation, as determined by electron microscopy, histology, immunohistochemistry, the terminal deoxynucleotidyl transferase-mediated nick endlabeling （TUNEL） method, reverse transcription-polymerase chain reaction. Functional effects of MG-132 on PMN accumulation, activation of nuclear factor kappa B （p65 mRNA and protein levels ）, and apoptosis were characterized in rat myocardial tissue. MG132 time-dependently inhibited myocardial p65 mRNA expression and reduced myocardial apoptotic index （AI） after reperfusion for 2 h, 6 h and 24 h （ P 〈 0. 01 ）. Moreover, MG-132 time-dependently decreased Bax protein levels, while increased Bcl-2 protein levels in ischemic and reperfused myocardium （ P 〈 0. 05 ）, its effect peaked after reperfusion for 24 h. Conclusions Our results demonstrate that MG-132 reduced myocardial reperfusion injury by inhibiting myosytic apoptotic cell death and blocking activation of NF-κB, down-regulating Bax expression and up-regulating Bcl-2 expression as well as el evating Bcl-2/Bax ratio.

Isobaric tags for relative and absolute quantitation-based quantitative proteomic analysis of X-linked inhibitor of apoptosis and H2AX in etoposide-induced renal cell carcinoma apoptosis

Background:X-linked inhibitor of apoptosis(XIAP)is a vital factor in the anti-apoptosis mechanism of tumors and is highly expressed in renal cell carcinoma(RCC).However,the mechanism through which XIAP regulates DNA damage repair is unknown.This study investigated the regulatory mechanism of XIAP in etoposide-induced apoptosis in two Caki-1 cell lines with high or low XIAP expression.Methods:The two cell lines were established using RNA interference technology.The differentially expressed proteins in the two cell lines were globally analyzed through an isobaric tags for relative and absolute quantitation-based quantitative proteomics approach.Proteomic analysis revealed 255,375,362,and 5 differentially expressed proteins after 0,0.5,3,and 12 h of drug stimulation,respectively,between the two cell lines.The identified differentially expressed proteins were involved in numerous biological processes.In addition,the expression of histone proteins(H1.4,H2AX,H3.1,H3.2,and H3.3)was drastically altered,and the effects of XIAP silencing were accompanied by the marked downregulation of H2AX.Protein-protein interactions were assessed and confirmed through immunofluorescence and Western blot analyses.Results:The results suggested that XIAP may act as a vital cell signal regulator that regulates the expression of DNA repair-related proteins,such as H2AX,and influences the DNA repair process.Conclusions:Given these functions,XIAP may be the decisive factor in determining the sensitivity of RCC cell apoptosis induction in response to chemotherapeutic agents.

Transarterial chemoembolization with PD-(L)1 inhibitors plus molecular targeted therapies for hepatocellular carcinoma(CHANCE001)

There is considerable potential for integrating transarterial chemoembolization(TACE),programmed death-(ligand)1(PD-[L]1)inhibitors,and molecular targeted treatments(MTT)in hepatocellular carcinoma(HCC).It is necessary to investigate the therapeutic efficacy and safety of TACE combined with PD-(L)1 inhibitors and MTT in real-world situations.In this nationwide,retrospective,cohort study,826 HCC patients receiving either TACE plus PD-(L)1 blockades and MTT(combination group,n=376)or TACE monotherapy(monotherapy group,n=450)were included from January 2018 to May 2021.The primary endpoint was progression-free survival(PFS)according to modified RECIST.The secondary outcomes included overall survival(OS),objective response rate(ORR),and safety.We performed propensity score matching approaches to reduce bias between two groups.After matching,228 pairs were included with a predominantly advanced disease population.Median PFS in combination group was 9.5 months(95%confidence interval[CI],8.4-11.0)versus 8.0 months(95%CI,6.6-9.5)(adjusted hazard ratio[HR],0.70,P=0.002).OS and ORR were also significantly higher in combination group(median OS,19.2[16.1-27.3]vs.15.7 months[13.0-20.2];adjusted HR,0.63,P=0.001;ORR,60.1%vs.32.0%;P<0.001).Grade 3/4 adverse events were observed at a rate of 15.8%and 7.5%in combination and monotherapy groups,respectively.Our results suggest that TACE plus PD-(L)1 blockades and MTT could significantly improve PFS,OS,and ORR versus TACE monotherapy for Chinese patients with predominantly advanced HCC in real-world practice,with an acceptable safety profile.

介入治疗动脉导管未闭109例临床观察

目的：观察介入治疗动脉导管未闭（PDA）近中期疗效，探讨介入治疗 PDA 的术后心脏结构改变情况及其安全性。方法回顾分析2010-2014年于遵义医学院附属医院心内科成功行经皮穿刺PDA介入封堵治疗患者109例，通过经胸超声心动图了解术前及术后3 d、1个月、3个月、6个月、1年心脏结构变化情况，通过并发症的发生情况评价其安全性。结果所有手术患者随访期内均健康生存，无介入治疗相关严重并发症。穿刺部位血管并发症发生2例（1.83%），术后残余分流发生9例（8.25%）。术后3 d超声心动图示收缩末期左心房内径（LAS）、左心室舒张期内径（LVD）、肺动脉内径（PA）较术前明显下降（P＜0.05）。左心室收缩期内径（LVS）术后1个月以后下降值具有统计学意义（P＜0.05）。≥15岁合并肺动脉高压患者术后3 d LAS、LVD、PA值与肺动脉平均压正常患者比较，下降更为明显（P＜0.05）。结论在严格掌握介入治疗适应证的情况下，经导管封堵治疗安全可靠，术后早期即可获益。

Individualized red-cell transfusion strategy for non-cardiac surgery in adults:a randomized controlled trial

Background:Red-cell transfusion is critical for surgery during the peri-operative period;however,the transfusion threshold remains controversial mainly owing to the diversity among patients.The patient’s medical status should be evaluated before making a transfusion decision.Herein,we developed an individualized transfusion strategy using the West-China-Liu’s Score based on the physiology of oxygen delivery/consumption balance and designed an open-label,multicenter,randomized clinical trial to verify whether it reduced red cell requirement as compared with that associated with restrictive and liberal strategies safely and effectively,providing valid evidence for peri-operative transfusion.Methods:Patients aged>14 years undergoing elective non-cardiac surgery with estimated blood loss>1000 mL or 20%blood volume and hemoglobin concentration<10 g/dL were randomly assigned to an individualized strategy,a restrictive strategy following China’s guideline or a liberal strategy with a transfusion threshold of hemoglobin concentration<9.5 g/dL.We evaluated two primary outcomes:the proportion of patients who received red blood cells(superiority test)and a composite of in-hospital complications and all-cause mortality by day 30(non-inferiority test).Results:We enrolled 1182 patients:379,419,and 384 received individualized,restrictive,and liberal strategies,respectively.Approximately 30.6%(116/379)of patients in the individualized strategy received a red-cell transfusion,less than 62.5%(262/419)in the restrictive strategy(absolute risk difference,31.92%;97.5%confidence interval[CI]:24.42-39.42%;odds ratio,3.78%;97.5%CI:2.70-5.30%;P<0.001),and 89.8%(345/384)in the liberal strategy(absolute risk difference,59.24%;97.5%CI:52.91-65.57%;odds ratio,20.06;97.5%CI:12.74-31.57;P<0.001).No statistically significant differences were found in the composite of in-hospital complications and mortality by day 30 among the three strategies.Conclusion:The individualized red-cell transfusion strategy using the West-China-Liu’s Score reduced red-cell transfusion without increasing in-hospital complications and mortality by day 30 when compared with restrictive and liberal strategies in elective non-cardiac surgeries.Trial registration:ClinicalTrials.gov,NCT01597232.

Bevacizumab biosimilar LY01008 compared with bevacizumab(Avastin)as first-line treatment for Chinese patients with unresectable,metastatic,or recurrent non-squamous non-small-cell lung cancer:A multicenter,randomized,double-blinded,phase Ⅲ trial

Background:Previous studies have demonstrated the preclinical pharmacological and toxicological consistency,and clinical pharmacokinetic equivalence of bevacizumab biosimilar LY01008 with reference bevacizumab(Avastin).This randomized controlled trial aimed to compare the efficacy and safety of LY01008 with Avastin in first-line treatment of Chinese patients with advanced or recurrent non-squamous non-small cell lung cancer(NSCLC).Methods:StageⅢB-ⅣNSCLC patients with evaluable lesions,good physical status,and adequate organ functions from 67 centers across China were randomized in a ratio of 1:1 to receive LY01008 or Avastin 15 mg/kg intravenously in combination with paclitaxel/carboplatin(combined treatment)for 4-6 cycles,followed by maintenance monotherapy with LY01008 until disease progression,intolerable toxicity,or death.The primary endpoint was objective response rate(ORR)in accordance with Response Evaluation Criteria in Solid Tumors(RECIST)version 1.1 confirmed by independent radiological review committees(IRRC).Secondary endpoints included disease control rate(DCR),duration of response(DoR),progression-free survival(PFS),overall survival(OS),and safety.This study was registered in Clinical Trials.gov(NCT03533127).Results:Between December 15^(th),2017,and May 15^(th),2019,a total of 649 patients were randomized to the LY01008(n=324)or Avastin(n=325)group.As of September 25th,2019 for primary endpoint analysis,589 patients received ORR evaluation,with a median number of combined treatment cycles of 5(range 1-6)andmedian duration of treatment of 3.0(range 0.0-5.1)months.ORRof responseevaluable patients in the LY01008 and Avastin groups were 48.5% and 53.0%,respectively.The stratified ORR ratio was 0.91(90%CI 0.80-1.04,within the prespecified equivalence margin of 0.75-1.33).Up to May 15^(th),2020,with a median follow-up of 13.6(range 0.8-28.4)months,no notable differences in DCR,median DoR,median PFS,median OS,and 1-year OS rate were observed between the LY01008 and Avastin groups.There were no clinically meaningful differences in safety and immunogenicity across treatment groups.Conclusions:LY01008 demonstrated similarity to Avastin in terms of efficacy and safety in Chinese patients with advanced or recurrent non-squamous NSCLC.LY01008 combined with paclitaxel/carboplatin is expected to become a new treatment option for unresectable,metastatic,LY01008 and Avastin groups.There were no clinically meaningful differences in safety and immunogenicity across treatment groups.Conclusions:LY01008 demonstrated similarity to Avastin in terms of efficacy and safety in Chinese patients with advanced or recurrent non-squamous NSCLC.LY01008 combined with paclitaxel/carboplatin is expected to become a new treatment option for unresectable,metastatic,or recurrent non-squamous NSCLC patients in the first-line setting.