Using phage-assisted continuous evolution (PACE) to evolve human PD1.

PD1/PDL1 pathway plays a critical role in cancer immune responses. The immune checkpoint inhibitors of PD1/PDL1 have been well explored and developed for immunotherapies of solid tumors. Recently, various monoclonal antibodies targeting the PD1/PDL1 pathway have emerged and achieved remarkable success in clinical trials. However, challenges with these monoclonal antibodies have appeared during cancer therapies, including predictors of response, patient selection, and innate resistance. Thus, a competitive antagonist of native PD1/PDL1, with smaller size and lower side-effect, is required for future cancer therapies. In this study, we utilized a protein evolution system of phage-assisted continuous evolution (PACE) to evolve PD1 continuously. Our results indicated that the newly evolved PD1 bound to PDL1 with higher affinity. The interactome analysis further suggested that these evolved PD1s exhibited higher specificity with PDL1. Therefore, these evolved PD1s may be applied as a new tool for tumor immunotherapy.

Is Propranolol Safe and Effective for Outpatient Use for Infantile Hemangioma? A Prospective Study of 679 Cases From One Center in China.

BACKGROUND: The protocol for the treatment of infantile hemangioma with propranolol varies among different clinical centers. METHODS: Six hundred seventy-nine patients who were 1 to 12 months old were recruited in this prospective study to receive propranolol treatment. The response to the propranolol therapy was classified as 4 levels. The results were primarily evaluated using color Doppler ultrasound examinations before and after propranolol treatment. RESULTS: The response was excellent in 176 (25.9%), good in 492 (72.5%), stable in 5 (0.7%), and poor in 6 (0.9%) of the patients. The mean age at the initiation of the therapy was 3.3 months (range, 1 to 10.9 months) and the mean duration of the therapy was 7.1 months (range, 3-17 months). The mean duration of the follow-up time after the discontinuation of the therapy was 5.3 months (range, 3-17 months). Regrowth of the hemangioma was observed in 92 cases (13.5%). Seventy-nine (11.6%) of the parents complained of their child's minor discomfort during the therapy. CONCLUSIONS: Propranolol (2 mg/kg per day) may significantly reduce the size of a hemangioma. As an outpatient therapy, propranolol was found to be safe for Chinese children and to have minor side effects.

Recurrence of infantile hemangioma after termination of propranolol treatment.

Propranolol has been the first-line treatment for problematic infantile hemangioma (IH) since 2008. The recurrence of IHs after stopping treatment with propranolol was reported in several studies. Mechanisms underlying this phenomenon have not been elucidated so far. Our study is the first to show the general pathology of recurrence of IH after stopping treatment with propranolol. It can provide help for the clinical treatment of IHs.

Celastrol alleviates airway hyperresponsiveness and inflammation in obese asthma through mediation of alveolar macrophage polarization.

Obese asthma is a unique asthma phenotype that decreases sensitivity to inhaled corticosteroids, and currently lacks efficient therapeutic medication. Celastrol, a powerful bioactive substance obtained naturally from the roots of Tripterygium wilfordii, has been reported to possess the potential effect of weight loss in obese individuals. However, its role in the treatment of obese asthma is not fully elucidated. In the present study, diet-induced obesity (DIO) mice were used with or without ovalbumin (OVA) sensitization, the therapeutic effects of celastrol on airway hyperresponsiveness (AHR) and airway inflammation were examined. We found celastrol significantly decreased methacholine-induced AHR in obese asthma, as well as reducing the infiltration of inflammatory cells and goblet cell hyperplasia in the airways. This effect was likely due to the inhibition of M1-type alveolar macrophages (AMs) polarization and the promotion of M2-type macrophage polarization. In vitro, celastrol yielded equivalent outcomes in Lipopolysaccharide (LPS)-treated RAW264.7 macrophage cells, featuring a reduction in the expression of M1 macrophage makers (iNOS, IL-1ß, TNF-α) and heightened M2 macrophage makers (Arg-1, IL-10). Mechanistically, the PI3K/AKT signaling pathway has been implicated in these processes. In conclusion, we demonstrated that celastrol assisted in mitigating various parameters of obese asthma by regulating the balance of M1/M2 AMs polarization.

The Effect of Transcutaneous Electrical Acupoint Stimulation on High-Risk Patients with PONV Undergoing Laparoscopic Gynecologic Surgery: A Randomized Controlled Trial.

BACKGROUND: Postoperative nausea and vomiting (PONV) is one of the most common complications after general anesthesia. The traditional comprehensive management of PONV usually uses one or two drugs, but this regimen fails to meet the requirements of the latest version of PONV guidelines. The purpose of this study was to evaluate the effect of transcutaneous electrical acupoint stimulation (TEAS) on high-risk PONV patients who are undergoing laparoscopic gynecological surgery. METHODS: In total, 162 high-risk PONV patients were randomly divided into an experimental group (n = 81) and a control group (n = 81). Both groups were injected with 4 mg of dexamethasone and 0.25 mg of palonosetron. In the experimental group, Nei-guan (PC6) and He-gu (LI4) were stimulated by a transcutaneous acupoint electrical stimulation instrument (HANS200E) 30 min before the surgery. The control group also received electrodes but no stimulation. Variance analysis and rank sum test were used to compare the differences between the two groups. RESULTS: The results of the incidence of postoperative nausea, vomiting, NRS score, degree of abdominal distension, and time to first flatus in the experimental group were lower than those in the control group. Nursing satisfaction of the experimental group was higher than that of the control group. CONCLUSIONS: The study demonstrates that TEAS combined with dexamethasone and palonosetron can effectively prevent PONV, reduce postoperative abdominal distension and postoperative pain, and shorten the first postoperative flatus time in high-risk patients with PONV. At the same time, it can improve nursing satisfaction.

Two-Z-epicanthoplasty in a three-dimensional model of Asian eyelids.

BACKGROUND: Epicanthoplasty is a procedure currently available for medial epicanthus correction. However, potential problems such as difficult designs, undercorrection, and unpleasant scarring in the medial canthus area make patients hesitant about undergoing epicanthoplasty. These barriers are challenges to surgeons. METHODS: From January 2007 to May 2010, epicanthoplasty was performed for 23 patients using two-Z-plasty in a three-dimensional surgery model. A total of 20 patients underwent a simultaneous double-eyelid operation when required. The medial canthal distance was measured preoperatively and 12 months postoperatively. The extent of postoperative scarring and improvement of the epicanthal fold were evaluated after surgery. RESULTS: The average intercanthal length decreased significantly from a mean of 37.0±2.1 mm preoperatively to 31.4±1.9 mm 12 months postoperatively (p<0.01, paired t test). Epicanthoplasty yielded excellent aesthetic results in terms of an open medial canthus without definite relapse, hypertrophic scarring, and injury of the lacrimal apparatus during the 12-month follow-up period. CONCLUSIONS: Two-Z-epicanthoplasty can be reproduced easily in a three-dimensional surgery model. This procedure is very effective for correction of the epicanthal fold, resulting in a pleasant-appearing, inconspicuous scar and a long-lasting outcome. LEVEL OF EVIDENCE IV: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors http://www.springer.com/00266.

Electroacupuncture prevents LPS- induced neuroinflammation via upregulation of PICK-TLR4 complexes in the microglia of hippocampus.

Sepsis-associated encephalopathy (SAE) is a common complication of sepsis caused by neuroinflammation. Electroacupuncture (EA) can be used to treat SAE, but the underlying mechanism is not clear. Lack of PICK1 further aggravates the inflammatory response in mice with sepsis. Therefore, we sought to investigate whether PICK1 is involved in the protective effects of electroacupuncture to SAE. In this study, mice were treated with EA after lipopolysaccharide (LPS) treatment. Behavioral tests; microglial activity of hippocampus; neuron survival and the inflammatory factors PICK1 and TLR4, as well as TLR4-related proteins, such as ERK, JNK, and P38, were assessed after EA treatment. PICK1, TLR4, and TLR4-related proteins, as well as PICK1-TLR4 complex levels were assessed in BV2 cells treated with LPS, PICK1 siRNA, or PICK1 polypeptide. The results indicated that EA could improve neurological assessment and reduce activation of microglial and TLR4 and expression of proinflammatory cytokines. EA also reduced the expression of TLR4 and phosphorylation of ERK/JNK/P38 while, increased the expression of PICK1 and TLR4 complexes. PICK1 knockdown further promoted the expression of TLR4 and phosphorylation of ERK/JNK/P38 in BV2 cells, but this effect was reversed by PICK1 polypeptides. These results suggest that EA may reduce neuroinflammation responses, decrease inflammatory factors, and finally, protect SAE by increasing the formation of PICK1-TLR4 complexes in microglia.

Ferroptosis is Involved in Hyperoxic Lung Injury in Neonatal Rats.

PURPOSE: To evaluate whether ferroptosis is involved in hyperoxic acute lung injury (HALI) and its mechanisms through the HALI model. METHODS: HE staining was used to assess lung injury pathology after the establishment of neonatal rat HALI model. ELISA was used to detect ROS, GPX4, and GSH expression. Prussian blue staining and Western Blot were used to detect iron deposition and the expression of ferroptosis-related proteins, respectively. RESULTS: The HALI group showed pathological changes with larger and fewer alveoli and thicker alveolar septa after HE staining. Prussian blue staining detected significant iron deposition in the lung tissue of the HALI group. GPX4, GSH, GSS, and SLC7A11 expressions were significantly decreased in the HALI group than in the normal control group. In contrast, ROS, TFRC, FHC, and FLC expressions showed opposite results (p<0.05). CONCLUSION: Ferroptosis may be involved in the pathological process of hyperoxic lung injury in neonatal rats.

Quantification of DNA methylation and hydroxymethylation in Alzheimer's disease mouse model using LC-MS/MS.

Ambiguous alteration patterns of 5-methylcytosine (5mC) and 5-hydroxymethylcytosine (5hmC) involved in Alzheimer's disease (AD) obstructed the mechanism investigation of this neurological disorder from epigenetic view. Here, we applied a fully quantitative and validated LC-MS/MS method to determine genomic 5mC and 5hmC in the brain cortex of 3 month-aged (12, 15, and 18 month) AD model mouse and found significant increases of 5mC and 5hmC levels in different months of AD mouse when compared with age-matched wild-type control and exhibited rising trend from 12-month to 18-month AD mouse, thereby supporting genomic DNA methylation and hydroxymethylation were positively correlated with developing AD.

[Topical timolol in the treatment of periocular superficial infantile hemangiomas: a prospective study].

OBJECTIVE: To investigate the efficacy and side effect of topical beta-blocker (Timolol Maleate) in the treatment of periocular hemangioma in a prospective study. METHODS: 432 outpatients with infantile hemangioma visited our special clinic service in Shanghai Ninth People's Hospital from July 2010 to December 2011. Among them, 12 superficial periocular lesions were selected in the study. Timolol was used topically on the lesion in every 12 hours. Two independent special doctors evaluated the results according to the pictures before and after four-week application of timolol. RESULTS: Were categorized into four levels: continuous growth (the lesion continues to grow), stable (no visible change), moderate (0-50% of regression) , perfect (more than 50% of improvement). Result of the 12 outpatients, 4 showed perfect result, 2 moderate, 4 stable and 2 continuous growth. No side effect was observed. CONCLUSIONS: Topical timolol is effective and safe in the treatment of superficial periocular infantile hemangioma. It could be considered as the first line treatment of proliferative superficial hemangioma.

[A prospective study of propranolol as first-line treatment for problematic infantile hemangioma in China].

OBJECTIVE: To prospectively assess the efficacy and safety or propranolol as a first-line treatment for problematic infantile haemangioma in China. METHODS: From Mar. 2009 to Feb. 2010, 78 patients with problematic infantile hemangioma were included in the prospective study. The characteristics of the tumor, including sex, age, site, complications, were recorded. The response to treatment at 1 week, at 1 month and at the end of treatment was evaluated. The efficacy of treatment was graded as no response, stabilization, or accelerated regression. The indications for treatment, side effects and relapse after treatment were documented. The mean follow-up period was 16.7 months (range, 12.1-23.6 months). RESULTS: Oral therapy was initiated at mean age of 3.7 months (range, 1.1-9.2 months) as first-line therapy. The mean age at the end of treatment was 11.2 months (range, 5.2-22.3 months). The treatment was lasted for 7.6 months (range, 2. 1-18.3 months). One week after treatment beginning, the hemangioma growth was controlled in all the patients. The accelerated regression was achieved in 88.5% (69/78) of patients after one week of treatment, and 98.7% (77/78) of patients after 1 month of treatment and at the end of treatment. Ulceration was occurred in 14 cases before treatment, which was healed after treatment for 2 months. Minor side effects were happened in 15.4% (12/78) of patients. Rebound growth of lesion was noticed in 35.9% (28/78) of patients. CONCLUSIONS: Propranolol is effective in the treatment of infantile hemangioma with minor side effect. We suggest it should be used as the first-line treatment.

[Clinical application of imiquimod for the treatment of infantile hemangiomas].

OBJECTIVE: To explore the clinical application of imiquimod for the treatment of infantile hemangiomas (IH). METHODS: 320 children with IH, including 250 superficial cases, 20 deep cases, and 50 mixed cases, were treated with 5% imiquimod cream every other day for 16 weeks. The clinical efficacy and side effects were evaluated at one year of age. RESULTS: The total effective rates of the superficial, deep, and mixed IH were 61.2% (153/250), 10.0% (2/20) and 60.0% (30/50) respectively, showing no statistical difference between superficial and deep type (P = 0.874), but significant difference between superficial and mixed (P < 0.01), deep and mixed type (P < 0.01). 56.0% (28/50) of mixed IH showed proliferation of its deep lesions. Slight skin erythema and crusting were the most common side effects. CONCLUSIONS: 5% imiquimod cream is effective and safe in superficial IH and superficial lesions of mixed IH with minimal skin reactions. The dysplasia of local tissue and systemic growth retardation are not found. It should be avoided to apply the cream to IH located around the cavities and skin fold. Imiquimod cream is a simple and convenient home-nursing medication. It can reduce care burden of family. Thus topical use of imiquimod can be considered as a good clinical indication for the treatment of superficial lesions of IH.