Cardiotoxicity is a serious complication of antineoplastic drugs. With the continuous development of antineoplastic drugs, immune checkpoint inhibitors have been used in the treatment of a variety of cancers. PD-1/PD-L1 immune checkpoint inhibitors have been widely concerned in recent years. This article reviews the manifestations and possible mechanisms of cardiotoxicity induced by PD-1/PD-L1 immune checkpoint inhibitors, and the detection methods and treatment of cardiotoxicity.

Objective To investigate the potential effects and mechanism on peroxisome proliferator-activated receptor γ-toll-like receptor 4-tumor necrosis factor-α (PPARγ-TLR4-TNF-α) targeted pathway on hyperglycemia induced myocardium inflammation and oxidative stress. Methods Thirty-two Japanese healthy adult rabbits were randomly divided into four groups with 8 rabbits in each group: normal control group (NC group), diabetes mellitus group (DM group), diabetes mellitus + pioglitazone 4 mg·kg-1·d-1 and 8 mg·kg-1·d-1 groups (DM+PGZ 4 mg and 8 mg groups). DM model was reproduced by alloxan of 150 mg/kg through auricular vein injection. On the same day of successful DM model reproduction, the diabetic rabbits were fed with corresponding dose of pioglitazone in DM+PGZ 4 mg and 8 mg groups, but the rabbits in NC group were not challenged. After 8 weeks of feeding, venous blood of left jugular vein bifurcation and myocardium tissue were harvested respectively for the determination of inflammation and oxidative stress parameters. TNF-α, interleukin-1 (IL-1), adiponectin (ADP), nitric oxide (NO) and total nitric oxide synthase (NOS) levels were determined by enzyme linked immunosorbent assay (ELISA), myeloperoxidase (MPO) activity was determined by colorimetric method, superoxide dismutase (SOD) activity was determined by hydroxylamine method, malondialdehyde (MDA) was determined by thiobarbituric acid colorimetric method, and catalase (CAT) activity was determined by UV spectrophotometry method. In addition, the mRNA expressions of TNF-α and TLR4 were determined by real-time quantitate reverse transcription-polymerase chain reaction (RT-qPCR). Results ① IL-1 and TNF-α in serum and myocardium of model rabbits were significantly increased, ADP was significantly decreased, and the mRNA expressions of TNF-α and TLR4 in myocardium were significantly increased, indicating a significant inflammatory reaction. The inflammatory reaction in pioglitazone intervention groups was significantly reduced, TNF-αand IL-1 levels in serum and myocardium of DM+PGZ 4 mg and 8 mg groups were significantly decreased as compared with those of DM group [serum: TNF-α(ng/L) was 268.33±46.57, 261.34±33.73 vs. 331.40±69.05, myocardium: TNF-α (ng/L) was 144.72±26.90, 139.59±14.59 vs. 177.48±27.40; serum: IL-1 (ng/L) was 24.40±2.56, 23.35±3.13 vs. 30.08±5.44, myocardium: IL-1 (ng/L) was 21.26±2.85, 20.54±2.75 vs. 24.78±3.60, all P < 0.05], and ADP levels were significantly increased [serum (μg/L): 19.64±8.85, 20.54±7.47 vs. 15.45±3.06, myocardium (μg/L): 10.31±2.22, 11.49±3.42 vs. 7.76±1.77, all P < 0.05], and the mRNA expressions of TNF-α and TLR4 in myocardium were significantly decreased (TNF-αmRNA: 0.15±0.05, 0.14±0.06 vs. 0.25±0.09; TLR4 mRNA: 0.57±0.17, 0.40±0.18 vs. 0.75±0.35, all P < 0.05). ②Oxidative stress in serum and myocardium of model rabbits was significantly increased, SOD, NO, and total NOS levels were significantly decreased while the serum CAT and MDA levels were significantly increased without effect on MPO. Compared with the DM group, SOD and NO levels in serum and myocardium were significantly increased in DM+PGZ 4 mg and 8 mg groups [serum: SOD (U/L) was 571.39±40.85, 609.28±54.47 vs. 535.10±37.08, myocardium:SOD (U/mg) was 55.74±8.12, 53.60±9.87 vs. 42.26±12.34; serum: NO (μmol/L) was 2.95±0.51, 2.99±0.43 vs. 2.03±0.78, myocardium: NO (nmol/mg) was 1.95±0.37, 2.11±0.26 vs. 1.56±0.33, all P < 0.05], the serum MDA levels were significantly decreased (μmol/L: 20.11±2.34, 19.70±2.02 vs. 23.07±3.06, both P < 0.05), while no significant effect on CAT. There was no significant difference in parameter of inflammatory and oxidative stress between the two pioglitazone intervention groups. Conclusion 4 mg·kg-1·d-1 pioglitazone could activate PPARγ-TLR4-TNF-α targeted pathway, thus inhibit inflammatory and oxidative stress factors expression, and down-regulate hyperglycemia induced myocardium inflammatory and oxidative stress level, but the effect did not show a dose dependent manner.

Objective To investigate changes in the number and function of endothelial progenitor cells (EPCs) in elderly patients with permanent atrial fibrillation(AF).Methods This prospective study included 35 elderly patients in each the permanent atrial fibrillation group and the control group.The numbers of circulating CD34+/KDR+ cells in the two groups were determined by flow cytometry.After two sets of peripheral blood samples were taken,mononuclear cells were isolated through density gradient centrifugation and cultured in vitro.EPC colonies were identified by the methylthiazolyldipheny-tetrazolium(MTT) assay and adhesion assay.The proliferation,adhesion and vasculogenesis of EPC colonies were determined by Matrigel culture.Enzyme-linked immunosorbent assay and nitric acid reductase assay were used to measure the secretion of vascular endothelial growth factor (VEGF) and nitric oxide (NO) in EPCs.Results The numbers of CD34+/KDR+ cells were lower in the AF group than in the control group (20.0±12.7)/104 vs.(77.9±58.9)/104 (P＜0.05).The number of EPC colonies in the atrial fibrillation group was significantly lower than that in the control group (1.8 ± 0.6) CFU vs.(3.5 ± 0.8) CFU (P ＜ 0.01).The proliferation,adhesion and vasculogenesis of EPC colonies in the AF group decreased,compared with the control group (each P＜0.01 or 0.05).Paracrine secretion of VEGF in the AF group (27.4±9.9)ng/L was lower than that in the control group (41.9±7.3)ng/L (P＜0.01) and paracrine production of NO in the AF group also decreased (P＜0.05).Conclusions EPCs In elderly patients with permanent atrial fibrillation show decreased numbers and reduced proliferation,adhesion and vasculogenesis.Paracrine VEGF and NO secretion is down as well.

OBJECTIVE To investigate the cytotoxic activity of Arca subcrenata Lischke anticancer protein(ASAP)constituents on human myeloid leukemia K562 cells in vitro and analyze its anticancer mechanisms. METHODS ASAP was extracted by low temperature water and ammonium sulfate precipitation. Protein concentration of ASAP was detected by Bradford method. Morphological changes of cultured K562 cells treated with ASAP were observed under the inverted phase-contrast micro?scope. The cell and nucleus changes were analyzed by Giemsa staining. The cytotoxicity of ASAP on K562 cells was detected by MTT assay. Flow cytometry was used to detect apoptosis and cell cycle of K562 cells treated with ASAP. The expression of apoptosis and cell cycle related proteins procaspase 3, caspase 3,P53 and programmed cell death 4(PDCD4)were analyzed by Western blotting. RESULTS ASAP exhibited significant cytotoxic effect on K562 cells in a time- and concentration-dependent manner. The concentration-effect correlation coefficient of ASAP 50,100 and 200 mg · L-1 on K562 cells for 24, 48 and 72 h was 0.851,0.8977 and 0.8997,respectively. Under an optical microscope,K562 cells showed cytomorphosis,or nuclear fragmentation after treatment with ASAP 200 mg · L-1 for 48 h. Flow cytometry analysis and Giemsa staining assay indicated that apoptotic cells increased and G2/M phase cells accumulated significantly with the increase of ASAP concentration. After treatment with ASAP 200 mg · L-1 for 48 h,the early and late apoptosis cell rate increased to(32.8 ± 0.1)%and(31.2 ± 2.2)%vs control group(3.7 ± 1.1)% and (9.9 ± 0.8)%(P<0.01),respectively,and the G2/M phase cells increased to (55.2 ± 1.7)% vs (15.3 ± 0.8)% in control group(P<0.01). After treatment with ASAP 200 mg · L-1 for 0-40 h,the expression of apoptotic protein procaspase 3 was down-regulated and its active form caspase 3 was significantly up-regulated at 32 h,while PDCD4 and P53 protein expression was down-regulated significantly in 0-40 h. CONCLUSION Apoptosis and cell cycle arrest induced in G2/M phase may account for ASAP cytotoxic activity to K562 cells. K562 cell apoptosis induced by ASAP depends on caspase 3 signal pathway. Down-regulated expression of PDCD4 and P53 proteins may be related to K562 cell apoptosis and cell cycle arrest in G2/M phase by ASAP.

Objective To explore the clinical efficacy and safety of modified Ponticelli regimen in treating patients with idiopathic membranous nephropathy (IMN).Methods A retrospective analysis was performed in 90 patients with IMN (type Ⅰ / Ⅱ,79/11 respectively) diagnosed by clinical data and renal biopsy.The patients were divided into modified Ponticelli group (n =23),steroid plus cyclophosphamide (CTX) (CTX group,n =39) and steroid plus cyclosporine A(CsA) (CsA group,n =28) according to the treatment.Liver function,renal function,serum lipid,proteinuria were recorded before and after treatment.Efficacy and adverse reactions were evaluated in three groups.Results (1) In all three groups,the quantity of proteinuria after treatment for 3 months [(3.33 ± 1.53) g/d,(4.70 ± 2.97) g/d,(3.92 ± 2.57) g/d],6 months [(1.60 ± 1.10) g/d,(2.34 ± 1.61) g/d,(2.25 ± 1.78) g/d] was significantly decreased compared with baseline level[(7.26 ± 2.06) g/d,(7.50 ± 2.55) g/d,(7.54 ± 2.70) g/d;P ＜ 0.05].Serum albumin levels at 3 months [(31.42 ± 3.86) g/d,(30.59 ± 5.79) g/d,(30.90 ± 7.87) g/d],6 months [(36.25 ± 4.20) g/d,(34.70 ± 6.70) g/d,(35.36 ± 8.29) g/d] were significantly increased compared with baseline levels [(24.13 ± 2.61) g/d,(23.98 ± 3.79) g/d,(22.94 ± 4.57) g/d;P ＜ 0.05],whereas serum creatinine at 3 and 6 months had no significant changes (P ＞ 0.05).(2) After treatment for 3 months,partial remission rates in modified Ponticelli group,CTX group and CsA group were 39.1％,35.9％,35.7％ respectively and complete remission rates were 8.7％,5.1％,10.7％,which were not statistically significant in all three groups (P ＞ 0.05).At 6 months,partial remission rates in three groups were 56.5％,41.0％,42.9％ respectively and complete remission rates were 21.7％,20.5％,28.6％,which did not suggested significant difference in all three groups either (P ＞ 0.05).(3) In modified Ponticelli group,steroid diabetes,impaired liver dysfunction,infections and gastrointestinal adverse events occurred in 1,1,2 and 2 patients,respectively.In CTX group,steroid diabetes,infections and gastrointestinal adverse events occurred in 5,8 and 2 patients,respectively.In CsA group,steroid diabetes and infections occurred in 1 and 3 patients,respectively.Conclusion Modified Ponticelli regimen to treat patients with IMN has a trend of better outcome than classic CTX regimen.The efficacy is not inferior to CsA regimen with fewer side effects.

Objective To investigate the effects of pioglitazone on atrial ionic channel remodeling in alloxan-induced diabetic rabbit models.Methods A total of 32 rabbits were randomly (random number) divided into control (CN) group,diabetes mellitus (DM) group,diabetes mellitus + pioglitazone 4 mg/ (d · kg) (DPG) group and diabetes mellitus + double pioglitazone 8 mg/ (d · kg) (DPI) group.The diabetic state was examined by quantitative determination of blood glucose levels of ≥ 14 mmol/L.Langendorff-perfused rabbit hearts were used to isolate single atrial myocyte,and whole-cell patch-clamp technique was used to record action potential duration (APD) and atrial ionic channel currents (ICa,L and INa).Variables with normal distribution were compared with One-way ANOVA and LSD-t test.Results Compared with controls,APD90 and APDS0 of left atrial myocytes were significantly prolonged in DM group (P ＜0.05 vs.CN),and there was no significant difference in APD90 frequency adaptation between them (P ＞0.05 vs.CN).The densities of INa were reduced and the densities of ICa,L were increased in DM group (P ＜ 0.01 vs.CN).The above variables were markedly attenuated in DPG and DPI group.Conclusions Pioglitazone may inhibits atrial ionic channel remodeling in diabetic rabbit models.

Objective To construct vectors expressing small interfering RNA targeting the Toll like receptor-4 (TLR4) gene and obtain TLR4 knock downed vascular smooth muscle cells (SMC). Methods Three small hairpin RNA (shRNA) targeting the TLR4 gene were designed, synthesized and cloned into the pSilence 2.1-U6 neo vector. Positive clones were verified with double enzyme digestion and sequencing. Then the recombinants were transfected to SMC by the cationic lipid method respectively.SMC were stably transfected with an expression plasmid and screened by G418. TLR4 mRNA and protein expression were detected by RT-PCR and Western blot methods. Results The pSilence2.1-siTLR4 ex-pression vectors were successfully constructed and a TLR4 knock-downed SMC cell line was established. RT-PCR and Western blot analysis confirmed that the expression of TLR4 was significantly down-regulated in the infected SMC cell line, and pSilence2.1-siTLR4-1was the most efficacious recombinant vector.Conclusion Recombinant vectors carrying shRNA targeting the TLR4 gene were successfully constructed and the TLR4 expression in vascular SMCs was inhibited.

Objective To explore the repair effect of mouse nerve growth factor (mNGF) on the neurological function of hand-foot-and-mouth disease (HFMD)children with central lesion. Methods 86 children with severe HFMD were divided into treatment group and control group ac-cording to the number of hospitalization,43 cases in each group.Both groups were treated with conventional comprehensive treatment for HFMD,while the treatment group was injected with mNGF on the basis of basic treatment.The disappearance time of symptoms associated with nervous system injury,neurological scores before treatment and on the third,seventh days of treatment as well as the levels of while cells and creatine kinase (CK)before and after treatment were compared, and the adverse reactions were observed.Results The disappearance time of vomiting,skittishness, limb shaking,myasthenia and somnolence as well as hospital stay in the treatment group were sig-nificantly shorter than those in the control group.Neurological scores on the third,seventh days of treatment decreased significantly in both groups,and neurological scores in treatment group were significantly lower than control group.After treatment,the levels of white cells and CK decreased significantly in both groups,and the decreased range in treatment group was larger.No obvious ab-normal reactions were observed by liver and kidney examination during treatment.Conclusion For children with HFMD and central lesion,application of mNGFcan effectively promote the repairmen of injured nervous tissue,improve the neurological function and shorten the course of disease.

Objective To explore the repair effect of mouse nerve growth factor (mNGF) on the neurological function of hand-foot-and-mouth disease (HFMD)children with central lesion. Methods 86 children with severe HFMD were divided into treatment group and control group ac-cording to the number of hospitalization,43 cases in each group.Both groups were treated with conventional comprehensive treatment for HFMD,while the treatment group was injected with mNGF on the basis of basic treatment.The disappearance time of symptoms associated with nervous system injury,neurological scores before treatment and on the third,seventh days of treatment as well as the levels of while cells and creatine kinase (CK)before and after treatment were compared, and the adverse reactions were observed.Results The disappearance time of vomiting,skittishness, limb shaking,myasthenia and somnolence as well as hospital stay in the treatment group were sig-nificantly shorter than those in the control group.Neurological scores on the third,seventh days of treatment decreased significantly in both groups,and neurological scores in treatment group were significantly lower than control group.After treatment,the levels of white cells and CK decreased significantly in both groups,and the decreased range in treatment group was larger.No obvious ab-normal reactions were observed by liver and kidney examination during treatment.Conclusion For children with HFMD and central lesion,application of mNGFcan effectively promote the repairmen of injured nervous tissue,improve the neurological function and shorten the course of disease.

Objective To explore the effect of clinical and pathological features on the incidence of Hyperuricemia (HUA) in renal glomerular disease.Methods A retrospective analysis was applied to review the clinical and pathological date collected from 3547 patients with renal glomerular disease.These patients were diagnosed as renal glomerular disease by renal biopsy from January 2007 to December 2011.Results (1) HUA incidence was 21.8％ (773/3547) in all of the patients,in which the incidence in secondary glomerular disease 27.2％ (240/882) was much higher than that in primary glomerular disease 20.7％ (552/2665),and the difference was significant (x2 =153.642,P ＜ 0.05).In primary glomerular disease,HUA incidence was the lowest in membranous nephropathy 14.4％ (96/665),while HUA incidence in lupus nephritis (LN) 45.3％(110/243) was the highest and small blood vessel infammation kidney damage 34.7％ (17/49) was the second in secondary glomerular disease.(2) With the increasing of glomerulosclerosis index,tubulointerstitial score,renal vascular lesions score and the stage of chronic kidney disease,HUA incidence increased (x2 =17.798-298.216,P =0.000).(3)Logistic regression analysis showed that high tubulointerstitial score,glomerulosclerosis index and renal dysfunction,male,overweight or obese,hypertension and hypertriglyceridemia were risk factors for hyperuricemia (OR:1.011-7.513,P ＜ 0.05).Conclusion The uric acid level is increased in nearly a quarter of patients with renal glomerular disease.Severe tubulointerstitial lesion,high glomerulosclerosis index,low glomerular filtration rate,male,overweight or obese,hypertension and hypertiglyceridemia were independent risk factors for HUA.