Objective To investigate the effect of intravitreal injection of fibrillin-2(FBN2)recombinant protein on FBN2-deficient retinopathy.Methods Thirty-two SPF-grade C57BL/6J mice were randomly divided into 4 groups:nor-mal control group,negative control group,FBN2 knockdown group,and FBN2 recombinant protein group,with 8 mice in each group.The right eyes were taken as the experimental eyes.Mice in the normal control group did not receive any inter-vention,mice in the negative control group were intravitreally injected with 3 μL empty vector(1 mg·L-1),and mice in the FBN2 knockdown group and FBN2 recombinant protein group were intravitreally injected with 3 μL adeno-associated vi-rus(1 mg·L-1).After 4 weeks,mice in the FBN2 recombinant protein group were intravitreally injected with 3 μL FBN2 recombinant protein(1 mg·L-1).Then,electroretinogram(ERG)and optical coherence tomography(OCT)were used to measure the amplitude of Rod-b and Max-a waves and the changes in the retinal structure.Real-time quantitative poly-merase chain reaction(RT-PCR)and Western blot were used to detect changes in FBN2,microfibril-associated glycopro-tein 2(MAGP-2),collagen I(COL1)mRNA and protein expression in the mouse retina.Results The ERG findings showed that compared with the negative control group and normal control group,the amplitude of Rod-b and Max-a waves in the retina of mice in the FBN2 knockdown group and FBN2 recombinant protein group decreased(all P＜0.05);com-pared with the FBN2 knockdown group,the amplitude of Rod-b and Max-a waves in the retina of mice in the FBN2 recom-binant protein group significantly increased(both P＜0.05).The OCT findings showed that compared with the FBN2 knock-down group,the structure of the retinal pigment epithelium and the light reflex in the FBN2 recombinant protein group be-came more regular.The RT-PCR detection results showed that compared with the FBN2 knockdown group,the expression of FBN2 mRNA in the retinal tissue of mice in the FBN2 recombinant protein group significantly increased,while the ex-pression of COL1 and MAGP-2 mRNA significantly decreased(all P＜0.05).Western blot assay results showed that com-pared with the FBN2 knockdown group,the expression of FBN2 protein in the retinal tissue of mice in the FBN2 recombi-nant protein group increased significantly,while the expression of COL1 and MAGP-2 proteins decreased significantly(all P＜0.05).Conclusion Intravitreal injection of FBN2 recombinant protein can compensate for the endogenous deficiency of FBN2 in mice with FBN2-deficient retinopathy and achieve therapeutic effects by regulating COL1 and MAGP-2 expres-sion.

Objective:To investigate the expression of latent transforming growth factor-β-binding protein (LTBP), transforming growth factor-β (TGF-β), cyclin-dependent kinase 2 (CDK2) and cyclin D2 (CCND2) in fibrillin-2 ( FBN2) interfering induced mouse retinopathy. Methods:Twenty-seven 8-week-old C57BL/6J mice were randomly divided into normal control group, empty vector group and FBN2 interference group according to the random number table method, with 9 mice in each group.The normal control group was not treated.The empty vector group and FBN2 interference group were intravitreally injected with 3 μl empty vector and 3 μl adeno-associated virus (AAV) carrying the sh-FBN2 interference plasmid in the right eye, respectively.The structural and functional changes of the retina were detected at 4 weeks after injection by optical coherence tomography (OCT) and full-field electroretinography (ERG).The expression and distribution of FBN2 protein in the retina were detected by immunofluorescence staining.The mRNA and protein expression levels of FBN2, LTBP-1, TGF-β2, CDK2 and CCND2 in mouse retina were detected by real-time fluorescence quantitative PCR and Western blot.All experiments complied with the ARVO statement.The research scheme was approved by the Experimental Animal Ethics Committee of Shandong University of Traditional Chinese Medicine (No.2019036).Results:Four weeks after injection, the results of OCT examination showed that compared with normal control and empty vector groups, the retinal pigment cortex of the FBN2 interference group was irregular with high density reflection areas.Full-field ERG results showed that compared with normal control and empty vector groups, the amplitude of Rod-a, Rod-b, Max-a and Max-b waveforms in FBN2 interference group decreased, and the differences were statistically significant (all at P<0.05).The results of immunofluorescence staining showed that FBN2 was expressed in the whole retina, and the fluorescence intensity of FBN2 was weaker in FBN2 interference group than that in normal control and empty vector groups.The fluorescence intensity of FBN2 in normal control group, empty vector group and FBN2 interference group was 16.21±2.21, 15.57±3.63 and 5.32±1.06, respectively, with a statistically significant overall difference ( F=66.03, P<0.05).The fluorescence intensity of FBN2 protein in FBN2 interference group was significantly lower than that in empty carrier group and normal control group (both at P<0.05).Compared with normal control and empty vector groups, the relative expression levels of LTBP-1 and TGF-β2 mRNA and protein were significantly increased in FBN2 interference group, while the relative expression levels of FBN2, CDK2 and CCND2 mRNA and protein were significantly decreased, and the differences were statistically significant (all at P<0.05). Conclusions:The increase of LTBP-1 and TGF-β2 and the decrease of G1/S phase related proteins CDK2 and CCND2 are involved in the development of FBN2-deficient retinopathy.

Objective:To evaluate the value of neutrophil to lymphocyte and platelet ratio (N/LPR) for predicting 28-day mortality in sepsis patients.Methods:A retrospective analysis was conducted. The clinical data of 154 sepsis patients admitted to intensive care unit (ICU) of the Affiliated Hospital of Jiangsu University from June 2017 to June 2020 were enrolled. The time of first diagnosis of sepsis in ICU was taken as the research starting point, and the death or 28 days as the end point. The 28-day outcomes of patients were recorded. The counts of peripheral blood neutrophil (NEU), lymphocyte (LYM) and platelet (PLT) were collected from all the enrolled patients within 3 days after diagnosis of sepsis. The ratios of N/LPR and NEU/LYM (NLR) were calculated respectively. The differences of N/LPR and NLR between survival group and death group were compared. Receiver operating characteristic (ROC) curve analysis was used to analyze the value of N/LPR and NLR on predicting the 28-day mortality of sepsis patients. According to the best cut-off value of ROC curve analysis, the 28-day mortality of patients with sepsis was analyzed by subgroup analysis, and the 28-day cumulative survival of patients with sepsis was analyzed by Kaplan-Meier survival curve.Results:Of the 154 sepsis patients, the patients with age < 18 years, pregnancy, blood disease, taking aspirin or other antiplatelet drugs within 1 week, taking leucocyte drugs within 1 week, length of ICU stay < 3 days and incomplete data were excluded. Finally, 50 patients were enrolled. Among them, 30 patients survived on the 28th day and 20 died. Compared with the survival group, the levels of N/LPR and NLR in the death group were significantly increased (N/LPR: 23.85±11.99 vs. 12.41±5.25, NLR: 17.83±8.69 vs. 10.75±3.63), with statistical differences (both P < 0.01). ROC curve analysis indicated that the area under ROC curve (AUC) of N/LPR for predicting 28-day death of sepsis patients was 0.827, it was higher than that of NLR (AUC = 0.762). Base on N/LPR≥15.48 as a predictor of cut-off value of death in 28 days of sepsis patients, the sensitivity was 75.0% and the specificity was 80.0%, respectively. Base on NLR≥10.65 as a predictor of cut-off value of death in 28 days of sepsis patients, the sensitivity was 75.0% and specificity was 56.7%, respectively. Subgroup analysis showed that the 28-day mortality in the patients with N/LPR≥15.48 ( n = 21) was significantly higher than those with N/LPR < 15.48 ( n = 29; 71.4% vs. 17.2%, χ 2 = 14.901, P < 0.01); and the 28-day mortality in the patients with NLR≥10.65 ( n = 28) was also significantly higher than those with NLR < 10.65 ( n = 22; 53.6% vs. 22.7%, χ 2 = 4.884, P < 0.05). The results were consistent with Kaplan-Meier survival curve analysis. Conclusion:Peripheral blood N/LPR has a good predictive value for 28-day mortality of sepsis patients, and which is better than NLR.

Objective To investigate the effect of laparoscopic cholecystectomy (LTCBDE) in treatment of patients with secondary extrahepatic bile duct stones. Methods Eighty-seven cases of our hospital patients with secondary to extrahepatic bile duct stones were randomly divided into the laparoscopic bile duct exploration and T tube drainage surgery (LCBDE) treatment group and the laparoscopic transcystic duct exploration of common bile duct lithotomy (ltcbde) treatment group. The observation focused on the operation time, bleeding volume , postoperative transfusion , postoperative drainage time , postoperative hospitalization time , cost of hospitalization, postoperative recovery time and complications compared clinical efficacies. Results LTCBDE group of patients in operation time (2.1 ± 0.5) was longer than that of the control group (1.6 ± 0.4), (P <0.001), while the bleeding volume, postoperative fluid volume, postoperative drainage time, postoperative hospitalization time, hospitalization expenses and postoperative recovery time were (17.4 ± 5.4), (6 550.4 ± 1 076.9), (3.5. 1.6), (4.1 ± 1.7), (12 243.5 ± 2 379.6), (11.3 ± 3.5) were lower than that of the group LCBDE (22.1 ± 7.5), (8 304.2 ± 1 394.8), (32.9 ± 10.4), (6.4 ± 2.4), (14 098.1 ± 2 897.3), (16.1 ± 5.7) P, respectively (P values were defined as 0.001, 0, 0, 0.015, 0.001, 0 individually); LTCBDE group of patients with bile leakage, acute peritonitis rates were 1/46,1/46,in which those were lower than the corresponding LCBDE in 6/41, 7/41 (P values were 0.033, 0.016, separately). Conclusion According to indications, LTCBD surgery has the advantages of less injury, less cost, less complications and so on. It has important significance to improve the condition of patients with secondary extrahepatic bile duct stones.

OBJECTIVETo explore the effects of exogenous carbon monoxide-releasing molecules 2 (CORM-2) on LPS-induced abnormal activation of platelets in peripheral blood of healthy human donors and its possible molecular mechanism.

METHODSVenous blood samples were collected from a healthy volunteer, and platelet-rich plasma (PRP) from the blood were isolated by differential centrifugation. The PRP was subpackaged into siliconized test tubes and then divided into control group, LPS group, inactive CORM-2 (iCORM-2) group, 10 µmol/L CORM-2 group, and 50 µmol/L CORM-2 group according to the random number table, with 3 tubes in each group. The PRP in control group did not receive any treatment. The PRP in LPS group received LPS (20 mL, 10 µg/mL) stimulation, and the PRP in iCORM-2 group, 10 µmol/L CORM-2 group, and 50 µmol/L CORM-2 group underwent the same LPS stimulation and treatment of 50 µmol/L iCORM-2, 10 µmol/L CORM-2, and 50 µmol/L CORM-2, respectively, with the dosage of 20 mL. After being cultured for 30 min, the platelet adhesion rate was determined by glass bottle method, the number of platelet spreading on fibrinogen was determined with immunofluorescent method, and the platelet aggregation rate was measured by turbidimetric method. The platelet poor plasma (PPP) was prepared from PRP, the levels of ATP in PPP and platelets were determined by chemical fluorescein method. The expressions of platelet glycoprotein I bα (GPIbα) and GPVI were analyzed by flow cytometer. The expressions of glycogen synthase kinase 3β (GSK-3β) and phosphorylated GSK-3β were determined by Western blotting and immunoprecipitation, respectively. Measurement of the above indices was repeated for 3 times. Data were processed with one-way analysis of variance and SNK test.

RESULTSCompared with those in control group, the platelet adhesion rates, numbers of platelets spreading on fibrinogen, platelet aggregation rates, expressions of GPIbα and GPVI in PRP, levels of ATP in PPP in LPS and iCORM-2 groups were significantly increased, while levels of ATP in platelets were significantly decreased (with P values below 0.05). Compared with those in LPS group, the former 7 indices in iCORM-2 group showed no significant differences (with P values above 0.05), while the levels of ATP in platelets in the 10 µmol/L CORM-2 and 50 µmol/L CORM-2 groups were significantly increased, and the other 6 indices in 10 µmol/L CORM-2 and 50 µmol/L CORM-2 groups were significantly decreased (with P values below 0.05). The expression levels of GSK-3β of the platelets in PRP in control, LPS, iCORM-2, 10 µmol/L CORM-2, and 50 µmol/L CORM-2 groups were 0.550 ± 0.060, 1.437 ± 0.214, 1.210 ± 0.108, 0.720 ± 0.010, and 0.670 ± 0.010, respectively, and the expression levels of the phosphorylated GSK-3β of the platelets in PRP in the above 5 groups were 0.950 ± 0.070, 1.607 ± 0.121, 1.420 ± 0.040, 1.167 ± 0.015, and 0.513 ± 0.122, respectively. Compared with those in control group, both the expression levels of GSK-3β and phosphorylated GSK-3β of the platelets in PRP in LPS and iCORM-2 groups were significantly increased (with P values below 0.05). The expression levels of GSK-3β and phosphorylated GSK-3β of the platelets in PRP between LPS group and iCORM-2 group were similar (with P values above 0.05). The expression levels of GSK-3β and phosphorylated GSK-3β of the platelets in PRP in 10 µmol/L CORM-2 and 50 µmol/L CORM-2 groups were significantly decreased compared with those in LPS group (with P values below 0.05).

CONCLUSIONSLPS stimulation can abnormally activate the platelets in peripheral blood of healthy human, but the abnormal activation can be inhibited by CORM-2 intervention, and the mechanism of the latter may involve the phosphorylation of GSK-3β mediated by GP.

Blood Platelets ; drug effects ; metabolism ; Carbon Monoxide ; metabolism ; Glycogen Synthase Kinase 3 ; Glycogen Synthase Kinase 3 beta ; Humans ; Lipopolysaccharides ; pharmacology ; Organometallic Compounds ; pharmacology ; Phosphorylation ; drug effects ; Platelet Activation ; drug effects ; Platelet Aggregation ; drug effects ; Platelet-Rich Plasma

Objective To evaluate the effect of Tirofiban on CRP levels in patients with acute myocardial infarction (AMI) after primary emergency percutaneous coronary intervention (PCI). Methods Eighty-four AMI patients admitted on emergency were randomly divided into two groups: (1) early-treated group (n=45), immediately receiving Tirofiban intravenously on admission and (2) late-treated group (n=39), receiving Tirofiban intravenously after coronary angiography was performed. TIMI grading before and after PCI in beth groups were compared, CRP levels before and three days after PCI were estimated. The major adverse cardiovascular events (MACEs) occurred during hospitalization and following-up period of three months were recorded. Results Before PCI, TIMI grade 3 forward flow rate in early-treated group was significantly higher than that in late-treated group, while no significant difference existed between two groups after PCI. Three days after PCI, CRP level in early-treated group was markedly lower than that in late-treated group. During hospitalization, the occurrence of MACEs in early-treated group was lower than that in late-treated group, while no marked difference was found between two groups during the following-up period of three months. Conclusion In treating AMI patients with primary PCI, Tirofiban should be used as early as possible, which is safe and effective for PCI and can also significantly improve forward blood flow in target vessels, decrease the ClIP level and reduce the occurrence of MACEs during hospitalization.

Recently,the relationship between psoriasis and coronary heart disease(CHD)have been casting close attention.It has been demonstrated that the prevalence of CHD in patients with psoriasis is more than one time higher than that of healthy people. Meanwhile,the prevalences of hypertension,diabetes mellitus, hyperlipoidemia and smoking in patients with psoriasis are higher than those of normal people,and some drugs for treatment of psoriasis may induce CHD.Therefore,the relationship between psoriasis and CHD is still unclear.In this paper,two cases of psoriasis complicated with CHD are reported,and the relationship between psoriasis and CHD is explored.

An old male patient visited the hospital due to shortness of breath and palpitation for 6 h, with fever 3 days before and pump failure at admission. Having no risk factor of coronary diseases such as hypertension, diabetes mellitus and obesity, with ST-T changes and abnormal Q wave on ECC, the signs were compatible with those of acute anterior wall myocardial infarction, while the characteristics of cardiac biomarkers ( significant increase in Troponin I and creatine kinase's isoform, and normal creatine kinase) were not in accordance with those of acute myocardial infarction. Emergency angiography was performed, which indicated normal coronary artery, normal pulmonary artery and global systolic dysfunction of left ventricle. The diagnosis of acute severe myocarditis was established, and intra-aortic balloon pump (IABP) was employed to provide hemodynamic support. Severe myocarditis mimicking acute myocardial infarction may be fatal, and can be easily misdiagnosed. Careful analysis of clinical manifestations, early diagnostic angiography and possible IABP placement are important for the successful treatment.

Objective To compare the effects of higher dosage of atorvastatin alone and combination of lower dosage of atorvastatin with ezetimibe on levels of lipid and metalloproteinases in patients with coronary artery disease. Methods Forty-two patients with coronary artery stenosis (50% to 70% stenosis) without stent replacement were selected, and were randomly divided into atorvastatin group (treated with 20 mg or 40 mg atorvastatin alone, n = 19) and combined-therapy group(treated with 5 mg, 10 mg atorvastatin and 10 mg ezetimibe, n =23). The levels of serum lipid, hepatic and renal functions, creatine kinase, metalloproteinase (MMP)-2, MMP-9 and tissue inhibitor of MMP-1 (TIMP-1) were detected 12 weeks after treatment. Results Twelve weeks after treatment, the level of LDL-C in atorvastain group was (1.94 ± 0.49) mmol/L (37.82% decrease from baseline), and that in combined-therapy group was (1.92 ±0.54) mmol/L(38. 26% decrease from baseline), and there was no significant difference between these two groups (P >0.05). Twelve weeks after treatment, the levels of MMP-2 and MMP-9 were significantly decreased and that of TIMP-1 was significantly increased compared with baseline levels in atorvastatin, while no such changes were detected in combined-therapy group. Conclusion In patients with coronary artery stenosis, combined therapy with lower dosage of atorvastatin and ezetimibe may lead to the same lipid-lowing effects compared with therapy with higher dosage of atorvastatin, but it may not result in a significant reduction in serum levels of MMP-2, MMP-9 and increase of TIMP-1.

Objective To investigate the effects of tirofiban application time on middle-term clinical prognosis in patients with acute ST segment elevation myocardial infarction (STEMI)treated by primary percutsneous coronary intervention (PCI). Methods The study of tirofiban was carried out in 50 patients with STEM[in cardiology department from January to December 2006. Twenty-nine patients were randomized to receive tirofiban after PCI for 24 - 36 hours(short time group, STG) and 21 patients for 48 - 72 hours (long time group,LTG). Clinical characteristics, angiography data, main adverse cardiovascular events (MACE) and coronary restenosis rate in 6-month follow-up of the two groups were. compared. Results Follow-up data showed that there was less intractable angina pectoris (14.3% vs 24.1%, P< 0.05) in LTG. But there was no significant difference in coronary restenosis rate between two groups. Conclusion Long time application of tirofiban following PCI in patients with STEMI could improve middle-term clinical prognosis by alleviating the incidence of intractable angina pectoris.