Preliminary clinical application of Keluoxin capsule for treatment of moderate and severe non-proliferative diabetic retinopathy / 中华眼底病杂志

Objective:To observe the safety and efficacy of Keluoxin capsules in the treatment of moderate to severe non-proliferative diabetic retinopathy (NPDR).Methods:An open-label, multi-center, single-arm, phase Ⅱa clinical trial. From May 2014 to December 2016, the patients diagnosed with moderate to severe NPDR who received Keroxin treatment in General Hospital of Central Theater Command, Affiliated Eye Hospital to Nanchang University, Xiyuan Hospital of China Academy of Chinese Medical Sciences, and Eye Hospital China Academy of Chinese Medical Sciences were divided into moderate NPDR group and severe NPDR group. The baseline data of the patients were obtained, best-corrected visual acuity (BCVA), optical coherence tomography, fundus fluorescein angiography and fundus photography were performed. On the basis of maintaining the original diabetes treatment, all patients took Keluoxin capsules orally for 24 weeks; 24 weeks after treatment was used as the time point for evaluating the efficacy. BCVA letters, central macular thickness (CMT) and 6 mm diameter total macular volume (TMV), retinal vascular leakage area, and retinal non-perfusion (RNP) area within an average diameter of 6 mm were compared between the two groups at baseline and 24 weeks after treatment. Independent sample Mann-Whitney U test was used to compare continuous variables between groups. Categorical data were compared by χ2 test. Results:A total of 60 NPDR patients and 60 eyes were included, 9 cases were lost to follow-up, and 51 cases and 51 eyes were finally included, including 37 eyes in the moderate NPDR group and 14 eyes in the severe NPDR group, respectively. At baseline, BCVA in moderate NPDR group and severe NPDR group were (80.1±6.8), (81.4±6.3) letters, respectively. CMT were (249.5±32.1), (258.9±22.2) μm, respectively. TMV were (8.79±1.09), (8.95±1.31) mm 3, respectively. Retinal vascular leakage areas were (7.69±10.63), (10.45±7.65) mm 2, respectively. RNP area were (2.48±5.74), (10.63±20.06) mm 2, respectively. There were 11 (29.7%, 11/37) and 4 (28.6%, 4/14) eyes with diabetic macular edema (DME), respectively; 24 weeks after treatment, BCVA in moderate NPDR group and severe NPDR group increased by (1.3±5.2), (3.2±3.0) letters, respectively. Compared with baseline, there was a statistically significant difference in the severe NPDR group ( t=-3.986, P=0.033). CMT were (252.1±45.6), (269.8± 57.2) μm, respectively. There were no significant differences compared with baseline ( t=-0.567, -0.925; P>0.05). TMV were (9.96±1.16), (10.09±1.32) mm 3, respectively. There were no significant differences compared with baseline ( t=-0.996, -1.304; P>0.05). Retinal vascular leakage area decreased (0.19±6.90), (1.98±7.52) mm 2, respectively. There were no significant differences compared with baseline ( t=0.168, 0.983; P>0.05). RNP area were (3.01±6.47), (10.36±19.57) mm 2, respectively. Compared with baseline, the differences were statistically significant ( t=-1.267, 0.553; P>0.05). There were 8 (21.6%, 8/37) and 3 (21.4%, 3/14) eyes with DME, respectively. Compared with baseline, the difference was statistically significant ( χ2=11.919, 4.571; P=0.001, 0.033). Conclusion:Keluoxin capsules can stabilize or improve BCVA, CMT, TMV and RNP area in patients with moderate and severe NPDR, and reduce the area of retinal vascular leakage.

The role and preliminary mechanism of Heparin-binding protein in the develoment of acute pancreatitis / 中华急诊医学杂志

Objective:To explore the role and preliminary mechanism of heparin-binding protein (HBP) in the development of acute pancreatitis (AP) through clinical analysis and animal models.Methods:(1) Clinical research: Blood samples were collected from AP patients admitted to the Second Affiliated Hospital of Anhui Medical University from January 1 to December 31, 2021 within 30 min of admission, including 20 patients with severe acute pancreatitis (SAP) and 20 patients with non-severe acute pancreatitis (NSAP). Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of HBP, syndecan-1 and hyaluronic acid (HA). Modified CT severity index (MCTSI), another 20 healthy volunteers were selected as controls (HC). Spearman correlation analysis was used to analyze the correlation between HBP and syndecan-1, HA and MCTSI. Receiver operating characteristic (ROC) curve was used to evaluate HBP to predict AP severity. (2)Animal experiment: The rat model of acute pancreatitis was prepared by intraperitoneal injection of L-arginine. In the normal control group (NC, n=8), the low molecular weight heparin (LMWH) intervention group ( n=8), and the acute pancreatitis group ( AP, n=8), the rats were euthanized 12 h later, and peripheral venous blood was collected to detect the levels of HBP, syndecan-1 and HA. Lung tissue and pancreas tissue were collected to observe the pathological damage, and the polysaccharide coating damage of vascular endothelial cells was observed under a fluoroscopy electron microscope. Results:The level of HBP at admission was significantly higher in the AP group than in the HC group, and the increase in the SAP group was more obvious. Correlation analysis showed that HBP was positively correlated with syndecan-1, HA and MCTSI. Animal studies found that the levels of HBP, syndecan-1 and HA in the AP group were significantly higher than those in the NC group. The pancreatic pathological score showed that the AP group was significantly increased, and the fluoroscopy electron microscope showed that the vascular polysaccharide coating was complete in the NC group, and the structure of the AP group was severely damaged. After LMWH intervention, the structure shedding and damage were significantly reduced, and the difference was statistically significant.Conclusions:HBP can promote the progression of AP, which is related to the destruction of the polysaccharide coating structure of endothelial cells and the increase of vascular permeability caused by HBP.

The protective effects of cyclosporin A on vascular permeability in sepsis rats / 药学学报

The protective effects of cyclosporin A (CsA), an inhibitor of mitochondrial permeability transition pore (MPTP), on vascular permeability in sepsis rats were investigated. Cecal ligation and puncture (CLP)-induced sepsis rats were used for in vivo studies, and the effects of CsA (1 and 5 mg·kg-1) on vascular permeability of lung, kidney, and intestine, mitochondrial respiratory control ratio, and the survival of the sepsis rats were observed. Lipopolysaccharide (LPS) was used for stimulating vascular endothelial cells (VECs) in vitro, and the effects of CsA on leakage of microvascular, immunofluorescence of zonula occludes-1 (ZO-1), and transendothelial electrical resistance (TER) were observed. All the animal welfare and experimental procedures are in accordance with the regulations of the Animal Ethics Committee of the Army Medical University. Compared with sham-operated group, the vascular permeability of lung, kidney, and intestine in sepsis rats increased significantly (P<0.05). Compared with conventional treatment group, CsA could significantly decrease the vascular permeability of lung, kidney, and intestine (P<0.05 or P<0.01), and prolong the survival period. The results of microcirculation also showed that CsA could significantly reduce the permeability of mesenteric venules in sepsis rats. At the cellular level, LPS stimulation significantly increased the permeability of vascular endothelial cells, including the decrease of transmembrane resistance and protein expression of ZO-1 (P<0.05). CsA can significantly reduce the increase of permeability of vascular endothelial cells induced by LPS stimulation (P<0.01). The function of mitochondria in the kidneys and intestines of sepsis rats was obviously impaired, and the respiratory control ratio of mitochondria was decreased. LPS significantly increased MPTP opening of VECs, while CsA significantly inhibited MPTP opening and improved mitochondrial function. CsA may protect mitochondrial function by inhibiting the opening of MPTP and play a protective role in the vascular permeability of sepsis rats. This study will provide an insight for the treatment of sepsis vascular leakage.

Arg-Leu-Tyr-Glu Suppresses Retinal Endothelial Permeability and Choroidal Neovascularization by Inhibiting the VEGF Receptor 2 Signaling Pathway

Vascular endothelial growth factor (VEGF) plays a pivotal role in pathologic ocular neovascularization and vascular leakage via activation of VEGF receptor 2 (VEGFR2). This study was undertaken to evaluate the therapeutic mechanisms and effects of the tetrapeptide Arg-Leu-Tyr-Glu (RLYE), a VEGFR2 inhibitor, in the development of vascular permeability and choroidal neovascularization (CNV). In cultured human retinal microvascular endothelial cells (HRMECs), treatment with RLYE blocked VEGF-A-induced phosphorylation of VEGFR2, Akt, ERK, and endothelial nitric oxide synthase (eNOS), leading to suppression of VEGF-A-mediated hyper-production of NO. Treatment with RLYE also inhibited VEGF-A-stimulated angiogenic processes (migration, proliferation, and tube formation) and the hyperpermeability of HRMECs, in addition to attenuating VEGF-A-induced angiogenesis and vascular permeability in mice. The anti-vascular permeability activity of RLYE was correlated with enhanced stability and positioning of the junction proteins VE-cadherin, β-catenin, claudin-5, and ZO-1, critical components of the cortical actin ring structure and retinal endothelial barrier, at the boundary between HRMECs stimulated with VEGF-A. Furthermore, intravitreally injected RLYE bound to retinal microvascular endothelium and inhibited laser-induced CNV in mice. These findings suggest that RLYE has potential as a therapeutic drug for the treatment of CNV by preventing VEGFR2-mediated vascular leakage and angiogenesis.

β₂-Adrenoceptor Blockade Deteriorates Systemic Anaphylaxis by Enhancing Hyperpermeability in Anesthetized Mice

PURPOSE: Patients treated with propranolol, a nonselective β-adrenoceptor antagonist, develop severe anaphylaxis, but the mechanism remains unknown. We determined effects of β₁- and β₂-adrenoceptor antagonists on the anaphylaxis-induced increase in vascular permeability in mice. METHODS: In anesthetized ovalbumin-sensitized C57BL mice, mean arterial blood pressure (MBP) was measured, and Evans blue dye extravasation and hematocrit (Hct) were assessed at 20 minutes after antigen injection. The following pretreatment groups (n=7/group) were studied: (1) sensitized control (non-pretreatment), (2) propranolol, (3) the selective β₂-adrenoceptor antagonist ICI 118,551, (4) the selective β₁-adrenoceptor antagonist atenolol, (5) adrenalectomy, (6) the selective β₂-adrenoceptor agonist terbutaline, and (7) non-sensitized groups. RESULTS: The antigen injection decreased MBP, and increased Hct and vascular permeability in the kidney, lung, mesentery, and intestine, but not in the liver or spleen. Pretreatment with ICI 118,551, propranolol and adrenalectomy, but not atenolol, reduced the survival rate and augmented the increases in Hct and vascular permeability in the kidney, intestine, and lung as compared with the sensitized control group. Pretreatment with terbutaline abolished the antigen-induced alterations. Plasma epinephrine levels were increased significantly in the sensitize control mice. CONCLUSIONS: Blockade of β₂-adrenoceptor can deteriorate systemic anaphylaxis by augmenting hyperpermeability-induced increase in plasma extravasation by inhibiting beneficial effects of epinephrine released from the adrenal glands in anesthetized mice.

Development of extravascular leakage of 6％ hydroxyethyl starch 130/0.4 when used for acute hemodilution in pediatric patients undergoing open heart surgery / 中华麻醉学杂志

Objective To evaluate the development of extravascular leakage of 6％ hydroxyethyl starch (HES) 130/0.4 when used for acute hemodilution in the pediatric patients undergoing open heart surgery.Methods Forty-eight American Society of Anesthesiologists physical status Ⅰ or Ⅱ pediatric patients,aged 2-12 yr,weighing 12-53 kg,scheduled for elective surgical repair of ventricular or atrial septal defect,were divided into 2 groups (n =24 each) according to age:preschool group (2 yr ≤ age ≤ 6 yr) and school-age group (6 yr＜age ≤ 12yr).After anesthesia induction and endotracheal intubation,a volume of 6％ HES 130/0.4 equivalent to 10％ of the blood volume was infused via the central veins at 0.5 ml · kg-1 · min-1 in two groups.Immediately before infusion of HES (T0) and at 15 and 30 min after the end of infusion (T1,2),blood samples were collected fron the central vein for determination of plasma colloid osmotic pressure (COP) and hemnoglobin (Hb) concentrations.The concentrations of 6％ HES 130/ 0.4 in plasma at T1 and T2 and in urine at T2 were measured by the anthranone colorimetric method.Results Compared with the baseline value at T0,the concentrations of Hh in plasma were significantly decreased at T1,2 in preschool group,and the concentrations of Hb in plasma were significantly decreased and plasma COP was increased at T1,2 in school-age group (P＜0.05).There were no significant differences in plasma Hb concentrations or COP at each time point between two groups (P＞0.05).The plasma 6％ HES 130/0.4 concentrations were significantly lower at T2 than at T1 in two groups (P＞0.05).Compared with school-age group,the plasma 6％ HES 130/0.4 concentrations were significantly decreased at T1,2 in preschool group (P＜0.05).There was no significant difference in 6％ HES 130/0.4 concentrations in urine between the two groups (P＞0.05).Conclusion When 6％ HES 130/0.4 is used for acute hemodilution,extravascular leakage happens after acute hemodilution and is more obvious in the preschool pediatric patients undergoing open heart surgery.

The role of heparin-binding protein in acute respiratory distress syndrome / 中华急诊医学杂志

Objective To study the role of heparin-binding protein (HBP) in sepsis-associated acute respiratory distress syndrome (ARDS),and to evaluate the prognostic value of HBP in ARDS.Method Sixty seven sepsis patients were enrolled in the prospective study.According to whether present ARDS,patients were divided into two groups:ARDS group and non-ARDS group.Blood samples were obtained within 2 hours after patients were diagnosed with sepsis.We measured the level of interleukin-6,interleukin8 and HBP by ELISA,counted the number of polymorphonuclear neutrophils (PMN),and calculated Acute Physiology and Chronic Health Evaluation Ⅱ score.We compared interleukin-6,interleukin-8,PMN and HBP between two groups by Student's t test.Correlation analysis was studied between HBP and other indicators by Spearman rank correlation coefficient.We also studied the early prognostic value on sepsisassociated ARDS by establishing receiver operating characteristic (ROC) curve.Results Interleukin-6 (t=4.25,P＜0.01),interleukin-8 (t=10.10,P＜0.01),PMN (t=3.47,P=0.0009) and HBP (t =0.0225,P =2.336,all showed significant difference between ARDS group and non-ARDS group.Interleukin-6 (r=0.535,P=0.002),interleukin-8 (r=0.419,P=0.017) and PMN (r=0.419,P =0.017) all had positive correlation with HBP.The area under curve (AUC) of HBP predicting ARDS was 0.796.Conclusions HBP level elevated in sepsis-associated ARDS patients and were correlated with interleuki-6,interleuki-8 and PMN.HBP show a high value on predicting emergence of sepsis-associated ARDS.HBP may play an important role in sepsis-associated ARDS pathological process.From this study we can conclude one of possible ARDS pathogenesis:inflammatory mediators,such as interleuki-6,interleuki8,activate PMN.Subseqoently PMN releases HBP.Then HBP resolts in vascolar leakage,which is one of the basic ARDS pathology.