Construction of airway organoid microinjection and polarity reversal model / 天津医药

Objective To explore novel methods for efficient respiratory viral infection of organoids by microinjection and polarity inversion techniques.Methods Lung tissue samples were obtained from 8-week-old male C57BL/6 mouse,and respiratory epithelial cells were extracted to establish a transwell organoid culture model.The green fluorescent protein(GFP)labeled influenza virus PR8(GFP-PR8)was quantitatively injected into organoids by improving the traditional microinjection platform,and morphologic changes in organoids and the immunofluorescence staining characteristics of tight junction proteins and microtubule proteins were observed.Polarity inversion apical-out(AO)was induced by suspension culture,and the morphological characteristics of polarity inversion was determined by HE staining.Normal and inverted organoids were infected with PR8,and the infection efficiency and expression differences of key pathway genes under different virus concentrations were observed.Results Ordinary organoids showed a significant increase in volume after microinjection.Following PR8 injection,the efficiency of infection was significantly higher in the apical region of organoids,accompanied by noticeable damage,as evidenced by significant down-regulation of tight junction proteins and microtubule protein expression.After suspension culture of the organoids,the polarity of ciliated cells gradually inverted outward over time,and the proportion of AO organoids stabilized on the 6th day.The efficiency of viral infection significantly increased in the inverted organoids,accompanied by significant cellular damage.After PR8 infection at 0.01 MOI,AO organoids showed significant changes in the inflammatory pathway and differentiation-related genes,with the opposite trend observed after higher concentration of PR8 infection.Conclusion Both polarity inversion and microinjection techniques significantly enhance the efficiency of influenza virus infection in organoids,thereby facilitating organoid widespread application in the field of respiratory tract infections.

Modeling the noninvasive bi-level positive airway pressure ventilation therapy system and simulated application / 生物医学工程学杂志

Without artificial airway though oral, nasal or airway incision, the bi-level positive airway pressure (Bi-PAP) has been widely employed for respiratory patients. In an effort to investigate the therapeutic effects and measures for the respiratory patients under the noninvasive Bi-PAP ventilation, a therapy system model was designed for virtual ventilation experiments. In this system model, it includes a sub-model of noninvasive Bi-PAP respirator, a sub-model of respiratory patient, and a sub-model of the breath circuit and mask. And based on the Matlab Simulink, a simulation platform for the noninvasive Bi-PAP therapy system was developed to conduct the virtual experiments in simulated respiratory patient with no spontaneous breathing (NSB), chronic obstructive pulmonary disease (COPD) and acute respiratory distress syndrome (ARDS). The simulated outputs such as the respiratory flows, pressures, volumes, etc, were collected and compared to the outputs which were obtained in the physical experiments with the active servo lung. By statistically analyzed with SPSS, the results demonstrated that there was no significant difference ( P > 0.1) and was in high similarity ( R > 0.7) between the data collected in simulations and physical experiments. The therapy system model of noninvasive Bi-PAP is probably applied for simulating the practical clinical experiment, and maybe conveniently applied to study the technology of noninvasive Bi-PAP for clinicians.

Rapid on-site pathogen diagnosis of intensive care unit: Microbiological Rapid On-Site Evaluation / 中华检验医学杂志

Sepsis-related deaths account for more than 50% of the total deaths in the world and are the primary problem threatening public health. Timely and rapid diagnosis is the key to improving the survival rate of patients with sepsis. Blood culture is a commonly used method to diagnose sepsis in clinics. However, it is time-consuming and unstable, which cannot used for pathogen quickly identification. In recent years, Microbiological rapid on-site evaluation (M-ROSE) based on morphological technique is applying to the clinics, showing significant advantages in timeliness and convenience, which would overcome the shortcomings of traditional diagnostic methods and be conducive to diagnosis of sepsis.

Distinct Transcriptional and Functional Differences of Lung Resident and Monocyte-Derived Alveolar Macrophages During the Recovery Period of Acute Lung Injury

In acute lung injury, two subsets of lung macrophages exist in the alveoli: tissue-resident alveolar macrophages (AMs) and monocyte-derived alveolar macrophages (MDMs).However, it is unclear whether these 2 subsets of macrophages have different functions and characteristics during the recovery phase. RNA-sequencing of AMs and MDMs from the recovery period of LPS-induced lung injury mice revealed their differences in proliferation, cell death, phagocytosis, inflammation and tissue repair. Using flow cytometry, we found that AMs showed a higher ability to proliferate, whereas MDMs expressed a larger amount of cell death. We also compared the ability of phagocytosing apoptotic cells and activating adaptive immunity and found that AMs have a stronger ability to phagocytose, while MDMs are the cells that activate lymphocytes during the resolving phase. By testing surface markers, we found that MDMs were more prone to the M1 phenotype, but expressed a higher level of pro-repairing genes. Finally, analysis of a publicly available set of single-cell RNA-sequencing data on bronchoalveolar lavage cells from patients with SARS-CoV-2 infection validated the double-sided role of MDMs. Blockade of inflammatory MDM recruitment using CCR2 −/− mice effectively attenuates lung injury. Therefore, AMs and MDMs exhibited large differences during recovery. AMs are long-lived M2-like tissue-resident macrophages that have a strong ability to proliferate and phagocytose. MDMs are a paradoxical group of macrophages that promote the repair of tissue damage despite being strongly pro-inflammatory early in infection, and they may undergo cell death as inflammation fades. Preventing the massive recruitment of inflammatory MDMs or promoting their transition to pro-repairing phenotype may be a new direction for the treatment of acute lung injury.

Protective effects of Slit2 on corneal epithelium and nerves in diabetic mice and its mechanism / 中华实验眼科杂志

Objective:To explore the promoting effects of slit guidance ligand 2 (Slit2) on the repair of corneal epithelium and nerve damage in diabetic mice and possible molecular mechanism.Methods:Sixty SPF C57BL/6 mice aged 5-6 weeks were divided into normal control group, diabetes model group and Slit2 injection group according to the random number table method, 20 for each group.Diabectic model was prepared by intraperitoneal injection of streptozotocin in the diabetes model group and Slit2 injection group.A mouse corneal epithelial injury repair model was established using electric epithelial scraper, and Slit2 recombinant protein was subconjunctivally injected immediately following modeling in the Slit2 injection group.The equal volume of phosphate buffer saline (PBS) was used in a same way in the diabetes model group.No intervention was performed in the normal control group.Corneal epithelial healing were examined at 24, 48 and 72 hours after corneal epithelial defect by corneal fluorescin staining.Real-time fluorescent quantitative PCR was used to detect the expression of Slit2 and its related receptors in the corneal epithelium of normal and diabetic model mice.Fluorescence staining of corneal wholemount with β-tubulin Ⅲ was used to observe the changes in corneal nerve morphology.Immunofluorescence staining was performed to detect the expression and distribution of Slit2 in mouse corneal epithelium in normal control group and diabetes model group, as well as the expression and distribution of Slit2, epidermal growth factor receptor (EGFR), extracellular-signal-regulated kinase (ERK), threonine protein kinase (AKT), β-catenin and Ki67 in the healing corneal epithelium of mice after corneal epithelium damage in different groups.The mouse corneal epithelial stem/progenitor cell line (TKE2) was divided into normal control group, high-glucose group and Slit2 treatment group.Western blot was performed to detect the expression of p-EGFR/EGFR and p-AKT/AKT in the TKE2 of the three groups.The expression of p-EGFR/EGFR and p-AKT/AKT in high glucose-cultured TKE2 with 0.01, 0.1 and 0.5 μg/ml Slit2 treatment for 10 minutes, and before and 10, 20, 30, 60, 120 minutes after 0.5 μg/ml Slit2 treatment was detected by Western blot.The effects of Slit2 on the axon regeneration of mouse trigeminal ganglion cells (TGs) were observed by immunofluorescence staining.The use and care of animals complied with the ARVO statement.This study protocol was approved by an Ethics Committee of Qingdao Eye Hospital of Shandong First Medical University (No.[2020]57).Results:At 48 and 72 hours after corneal epithelial scraping, the speed of corneal epithelial repair was significantly slowed down in diabetes model group in comparison with the normal control group and Slit2 injection group.The relative expression levels of Slit2 and its receptors Robo1, Robo2 and Robo4 mRNA in the normal corneal epithelium in the diabetes model group were significantly higher than those of the normal control group (all at P<0.05). The fluorescence intensity of Slit2 in normal corneal epithelium in diabetes model group was similar to the normal control group, and the fluorescence intensity of Slit2 in damaged corneal epithelium in diabetic mice was significantly weaker than that in normal control group.Corneal nerve plexus was denser at 7 days after corneal epithelial injury and the nerve fibers were increased with more branches in Slit2 injection group compared with diabetic group.The fluorescence intensity of p-EGFR, p-ERK, β-catenin and Ki67 in damaged corneal epithelium in normal control group and Slit2 injection group was stronger than that of the diabetes model group.The relative expression levels of p-EGFR/EGFR, p-AKT/AKT, and β-catenin in TKE2 in high-glucose group were significantly lower than those in normal control group and Slit2 treatment group (all at P<0.05). The relative expression levels of p-EGFR/EGFR and p-AKT/AKT in high glucose-cultured TKE2 after Slit2 treatment were significantly increased in comparison with before Slit2 treatment (both at P<0.05), and the relative expression levels of p-EGFR/EGFR and p-AKT/AKT in TKE2 were elevated as the increase of Slit2 concentration.The activation effect of 0.5 μg/ml Slit2 on EGFR and AKT pathways was most obvious.The synapse length of TGs cultured by high glucose was (40.52±5.44) μm, which was significantly shortened than (72.14±9.48) μm in normal control group and (73.04±4.66) μm in Slit2 injection group (both at P<0.05). Conclusions:Slit2 can protect the corneal epithelium by activating EGFR signaling pathway and play a protective role to neurons by increasing the density of corneal subepithelial plexus and promoting the growth of TGs axons in diabetic mice.

Role of NLRP3 inflammasome signaling pathway in the occurrence and development of diabetic keratopathy / 中华实验眼科杂志

Diabetic keratopathy is a chronic complication of diabetes caused by abnormal metabolites accumulation, oxidative stress, abnormal inflammation and corneal neuropathy.It can result in delayed corneal epithelial healing and decreased corneal sensitivity under the stimulation of ocular trauma or surgery which bring great challenges to clinicians.Activation of nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) inflammatory is one of the factors that cause chronic complications of diabetes, and is also an important factor for delaying the healing of diabetic wounds.The NLRP3 inflammatory signaling pathway is closely related to corneal oxidative stress, delayed epithelium healing and development of corneal neuropathy.In this paper, the research status and prospects of NLRP3 inflammatory signaling pathway and diabetic keratopathy were reviewed to provide new ideas for studying the mechanism and treatment of diabetic keratopathy.

Research of immunoregulation effects of regulatory T cells in dry eye disease / 中华实验眼科杂志

Dry eye disease is a multifactorial disease affecting the ocular surface, lacrimal glands and meibomian glands.Its incidence is gradually increasing and tends to occur in the younger.Its main features are ocular surface inflammation caused by tear film instability and high osmotic pressure of tears.Moreover, there is a vicious circle between inflammation and ocular surface damage.Immune-related inflammatory responses play a key role in this process.Regulatory T cell (Treg) is a subset of T cells with immunoregulatory functions, which are closely related to the occurrence and development of dry eye, and can inhibit the inflammation of dry eyes by acting on antigen-presenting cells and T helper cell (Th)1/Th17.Recent studies have shown that Treg in dry eye is abnormal in number or function and closely related to the risk factors of dry eye such as age and gender.In addition, by increasing the number of Tregs and promoting their differentiation to alleviate inflammatory response can provide new treatment strategies in dry eye.The correlation between Treg and dry eye and its related research in the pathogenesis and treatment of dry eye were reviewed in this article.

Role of mitophagy in diabetic corneal endothelial lesion / 中华实验眼科杂志

Diabetic keratopathy is one of the common ocular complications of diabetes, and diabetic patients are often accompanied by changes in the morphological structure of the corneal endothelium.Oxidative stress, inflammation, apoptosis, glucose metabolism disorders, mitochondrial injury, and endoplasmic reticulum stress are the main mechanisms of the occurrence and progression of diabetic keratopathy.Studies have shown that advanced glycation end products can activate and induce the formation of a large number of reactive oxygen species (ROS), which in turn causes cell damage and even apoptosis.Mitochondria are the source of ROS, which will be damaged when a large amount of ROS accumulate, and mitochondrial autophagy will be formed when the body removes damaged mitochondria.Mitophagy refers to the process of eliminating aging, dysfunctional, damaged mitochondria through selective autophagy, which is a key mechanism for mitochondria to maintain function.The decrease in the level of mitophagy will lead to the destruction of the hexagonal structure of the diabetic corneal endothelium and its dysfunction, and upregulating the level of mitophagy can play a protective role on corneal endothelium in oxidative stress.The role of mitophagy in diabetic corneal endothelial lesions were reviewed in this article.

Progress on basic and clinical research of immunology in lung transplantation / 器官移植

The quality of life of organ transplant recipients is closely correlated with immune status. Compared with those undergoing other solid organ transplantation, the long-term prognosis of lung transplant recipients is worse. The underlying immune mechanism is complex with both similarities and characteristics. Therefore, in-depth understanding of the immune mechanism in the process of immune response of allogeneic lung transplantation plays a critical role in improving the long-term survival of the recipients. In this article, the unique composition of immune cells in the lung, the characteristics of rejection after lung transplantation, the early warning and differential diagnosis of pathogen infection in lung transplantation and postoperative complications after lung transplantation were investigated. Research progress on clinical diagnosis and basic studies related to immunology in allogeneic lung transplantation were summarized, aiming to elucidate the immunological characteristics of lung transplantation and provide theoretical basis for improving the longterm survival of lung transplant recipients and prevention and treatment of allograft dysfunction.

Application of femtosecond laser in keratoplasty / 中华实验眼科杂志

Femtosecond laser has been widely used in ophthalmology for its high precision, safety and minimal damage to tissues, and now its application in the corneal refractive surgery, corneal stromal ring implantation and cataract surgery has been more mature.In recent years, it has been actively developed and applied in penetrating keratoplasty, anterior lamellar keratoplasty and corneal endothelial transplantation, and some achievements have been obtained in experimental research and clinical application.While femtosecond laser promotes the development of corneal transplantation surgery, there are still some problems such as limited indications, uncertain efficacy, expensive equipment, lack of big data support and intelligence in personalized design.Continuous optimization and improvement are still needed to ensure postoperative vision, reduce postoperative complications, simplify parameter design and reduce consumption costs.In this paper, the indications, parameter design, clinical effect and limitations of femtosecond laser in keratoplasty were reviewed.

Roles of dendritic cells in the occurrence and development of diabetic keratopathy / 中华实验眼科杂志

Diabetic keratopathy (DK) is a common ocular complication of diabetes.Long-term hyperglycemia impairs many structures of the cornea, leading to corneal opacity and visual dysfunction.A large number of researches focus on the epithelium and nerve abnormities in DK, but the pathogenesis is not completely elucidated.Dendritic cells (DCs) are specialized antigen-presenting cells, linking innate and adaptive immunity, participating in the occurrence and development of diabetes and its complications.To date, there are many myths in relationship between DCs and DK to be solved, and there are a few researches that investigate the relation between DCs and the occurrence and development of diabetes.In this article, the pathogenesis and pathogenic changes of DK, the types and functions of different DCs, the relationship between DCs and chronic inflammation and delayed healing of corneal epithelium in DK, as well as the role of DCs in corneal neuropathy were reviewed in order to provide some references for further investigations about the pathogenesis and treatment of DK.

Screening results of tumor markers CA-724, CA-153, CA-125, CA-199 and CEA in 45~60-year-old health examination population in Chengdu / 公共卫生与预防医学

Objective To analyze the screening results of tumor markers carbohydrate antigen 724 (CA-724), carbohydrate antigen 153 (CA-153), carbohydrate antigen 125 (CA-125), carbohydrate antigen 199 (CA-199) and carcinoembryonic antigen (CEA) in the 45 ~ 60-year-old health examination population in Chengdu area. Methods A total of 2 268 healthy people aged 45 ~ 60 years old living in Chengdu area from May 2019 to August 2020 were selected. Levels of serum tumor markers CA-724, CA-153, CA-125, CA-199 and CEA were detected by electrochemiluminescence. Positive rates of different tumor markers and confirmed diagnosis of malignant tumors were statistically analyzed. People with positive and negative tumor markers were respectively included in the positive group and the negative group, and their serum tumor markers were compared. The screening results of tumor markers in the health examination population of different gender and age, and the diagnostic efficiency of each tumor marker for malignant tumors were analyzed. Results Among the 2 268 healthy people, the positive rates of CA-724, CA-153, CA-125, CA-199 and CEA were 10.05%, 4.98%, 1.10%, 7.01% and 4.76%, respectively. A total of 72 cases were positive for tumor marker detection, with a total positive rate of 3.17% (72/2,268). Among them, 12 cases were diagnosed as early or mid-term tumors by pathology, imaging and other examinations, with a total tumor detection rate of 0.53% (12/2,268), including 8 cases of malignant tumors, with a detection rate of 0.35% (8/2,268). The levels of CA-724, CA-153, CA-125, CA-199 and CEA in the tumor markers positive group were higher than those in the negative group (P<0.05). The CA-125 and CA-199 positive rates in males were lower than those in females, while CEA positive rate was higher in males than in females (P<0.05). Positive rates of CA-153, CA-199 and CEA in different age groups showed statistically significant differences (P<0.05). Positive expression of CA-153 and CA-199 mainly occurred in the 51~60 years old group, and positive expression of CEA mainly occurred in the 45~50 years old group. ROC curve analysis showed that combined detection of CA-724, CA-153, CA-125, CA-199 and CEA had the highest sensitivity, specificity and accuracy in predicting malignant tumors, and the area under the curve was the largest (0.865). Conclusion Tumor markers CA-724, CA-153, CA-125, CA-199 and CEA are highly expressed in the 45 ~ 60-year-old health examination population in Chengdu area, and positive rate of CA-724 is the highest. Levels of tumor markers in people of different gender and age are also different. Joint detection of various tumor markers is conducive to early detection and treatment of the disease, and reduction of deterioration risk.

Advances in the relationship between insulin-like growth factor system and corneal diseases / 中华实验眼科杂志

Recently, an increasing number of studies on the insulin-like growth factor (IGF) system have been conducted.In addition to its well-known regulation of cell proliferation and differentiation, enhanced metabolic activity, anti-apoptosis, and promotion of cell survival, the role of IGF in tumor growth, autophagy, longevity and aging, and oxidative stress has become a hot spot.Drugs targeting IGF signaling pathways are also progressing into the clinical experimental phase.This article summarized the important role of IGFs and its major signaling pathways in the growth and development, their relationship with the corneal diseases, and discussed the related problems and prospects.

Role of the nuclear factor kappa B signaling pathway in the pathogenesis and development of diabetic keratopathy / 中华实验眼科杂志

In recent years, with the increasing incidence of diabetes, diabetic keratopathy has gradually attracted more attention from clinicians.Diabetic keratopathy is mainly manifested as delayed corneal epithelial healing, corneal edema, decreased corneal sensitivity, and neurotrophic corneal ulcer.There are many mechanisms of diabetic keratopathy, including the abnormal accumulation of metabolites, an inflammatory reaction, oxidative stress, the lack of neurotrophic factors, corneal limbal stem cell abnormalities, etc.At present, most of the research on diabetic keratopathy at home and abroad has mainly focused on epithelial damage repair and changes in nerve function, and the pathological mechanism is mainly inflammatory response and apoptosis.Nuclear factor kappa B(NF-κB) is an important transcription regulatory factor, and is involved in regulating the expression of inflammatory factors, chemokines, cell adhesion molecules, apoptosis-related genes, growth factors, and other target genes.Studies have reported that chronic inflammation and epithelial cell apoptosis of the diabetic corneal epithelium are involved in the regulation of the NF-κB signaling pathway, which is also involved in the regulation of diabetic peripheral neuropathy, and diabetic corneal neuropathy belongs to the category of diabetic peripheral neuropathy.The NF-κB signaling pathway is closely related to the occurrence and development of diabetic keratopathy, and this article summarizes its mechanism.

Analysis of pathogenic factors and etiological characteristics of 531 patients with suppurative endophthalmitis / 中华眼底病杂志

Objective To analyze pathogenic factors and etiological characteristics of suppurative endophthalmitis.Methods A total of 531 consecutive patients (531 eyes) with suppurative endophthalmitis who were hospitalized in Qingdao Eye Hospital of Shandong Eye Institute from January 2006 to December 2015 were included in the study.Among them,410 patients with 410 eyes were males (77.2％),121 patients with 121 eyes were females (22.8％).The average age of the patients was 38.62± 15.36 years.The relevant medical records were collected to analyze the pathogenic factors.Samples of aqueous humor,vitreous or other intraocular samples were taken under aseptic conditions for bacterial and fungal culture and in vitro drug sensitivity test.Results Ocular trauma was the primary pathogenic factor of suppurative endophthalmitis (60.1％),other factors included postoperative endophthalmitis (19.0％),suppurative keratitis-related endophthalmitis (17.1％) and endogenous endophthalmitis (3.8％).Postoperative endophthalmitis mainly occured after cataract surgery.A total of 224 strains of organisms were isolated,among which the predominant organisms isolated were gram-positive bacteria (54.0％) and staphylococcus epidermidis was the most common (25.0％).The other pathogenic organisms were fungi (29.5％) and gram-negative bacteria (16.5％).Among the fungi,aspergillus (10.7％) was the dominant genus,followed by fusarium (9.8％).For gram-positive organisms,susceptibilities were vancomycin 97.4％,gatifloxacin 91.8％,fusidate acid 77.9％ and levofloxacin 54.6％.For gram-negative organisms,susceptibilities were gatifloxacin 85.7％,levofloxacin 77.8％,tobramycin 71.4％ and ceftazidime 62.5％.For fungal isolates,sensitivities were voriconazole 88.2％ and amphotericin B 84.8％.Conclusions Ocular trauma is the main pathogenic factor of suppurative endophthalmitis,followed by postoperative endophthalmitis and suppurative keratitis-related endophthalmitis.Gram-positive bacteria are the major pathogenic organisms,especially staphylococcus epidermidis followed by fungal species,among which aspergillus and fusarium were the dominating pathogenic genus.

Therapeutic effects of different doses of methylprednisolone on smoke inhalation-induced acute lung injury in rats / 中华危重病急救医学

Objective To investigate the therapeutic effect of different doses of methylprednisolone (MP) in smoke inhalation-induced acute lung injury (SI-ALI).Methods Adult male Sprague-Dawley (SD) rats were divided into control group (group A,n = 6), smoke inhalation group (group B, smoke inhalation 30 minutes,n = 30) and smoke+MP 40, 4, 0.4 mg/kg intervention group (groups C, D, E; intraperitoneal injection of MP at 1 hour before smoke inhalation, n = 30) according to random number table method. The survival status of rats in each group was observed at 24 hours, and murine smoke inhalation induced trauma score (MSITS) according to the symptoms and signs of rats at 3 hours after smoke inhalation were scored. The blood of abdominal aorta of rats was collected. Then the rats were sacrificed to harvest bronchoalveolar lavage fluid (BALF) and lung tissue. The levels of interleukin (IL-6, IL-17a) in plasma and BALF were detected by enzyme linked immunosorbent assay (ELISA); the total number of white blood cells and the proportion of leukocytes or macrophages in BALF were calculated; the histopathological changes of lung were observed and the lung injury score was given; the expression of myeloperoxidase (MPO) and high mobility group protein B1 (HMGB1) in lung tissue were detected by Western Blot.Results The 24-hour survival rate of group B rats was 33.67％. The survivalrate of groups C, D and E (65.73％, 85.17％, 60.07％) were significantly higher than that of group B (allP < 0.05), and the survival rate of group D was significantly higher than that of groups C and E. Diffuse inflammatory cell infiltration, intra-alveolar hemorrhage and a large amount of edema fluid were seen in the lung tissue of group B; and the lung injury score was significantly higher than that of group A. Compared with group B, the lung injury in different doses of MP group were decreased to different degrees, while the lung injury scores in groups C and D were significantly decreased (3.31±1.37, 2.62±0.98 vs. 5.52±0.97, bothP < 0.01); correlation analysis showed that MSITS score was significantly and positively correlated with lung injury score (r = 0.862,P < 0.001). The levels of plasma inflammatory factors and BALF protein, inflammatory cells and inflammatory factors, and the expression of MPO, HMGB1 in group B were significantly higher than those in group A. Compared with group B, the levels of inflammatory factors in plasma, and protein content, inflammatory cells and inflammatory factors in BALF in different doses of MP group were decreased to different degrees, with significant differences in groups C and D [plasma: IL-17a (pg/L): 49.28±27.12, 36.57±16.52 vs. 191.79±88.21; IL-6 (ng/L): 206.47±109.96, 197.52±113.86 vs. 669.00±299.60; BALF: protein content (mg/L):892.0±164.5, 566.1±120.9 vs. 1838.0±145.8; white blood cell count (×109/L): 5.40±1.67, 2.81±1.20 vs. 9.02± 2.06; neutrophil ratio: 0.315±0.081, 0.273±0.080 vs. 0.590±0.096; IL-17a (ng/L): 22.63±8.62, 18.92±8.43 vs. 43.31±19.17; IL-6 (ng/L): 156.49±46.94, 123.66±64.91 vs. 253.43±80.03; allP< 0.01]; in addition, the expression of MPO and HMGB1 protein in lung tissues of MP groups with different doses were significantly decreased, the expression of MPO in group D was significantly lower than that in group E [MPO/β-actin (fold increase from group A):2.14±0.97 vs. 4.35±0.87,P < 0.01], the expression of HMGB1 in groups C and D were significantly lower than that in group E [HMGB1/β-actin (fold increase from group A): 1.77±0.73, 1.23±0.67 vs. 3.65±1.08, bothP < 0.05]. Conclusions MP can significantly improve the survival rate of SI-ALI rats and reduce the acute pulmonary and systemic inflammatory response. The MP effect of 4 mg/kg was better than 40 mg/kg and 0.4 mg/kg.

Research advances in toxic keratopathy / 眼科新进展

The incidence of toxic keratopatby is increasing yearly.Due to the shortage of specific clinical manifestations,the diagnosis and treatment of the disease are relatively difficult,hence toxic keratopathy has received extensive attention of the domestic and abroad scholars.In recent years,with the development of ophthalmic examination technology and progress of instruments,its mechanism,diagnosis and treatment have been newly cognized.These three aspects will be reviewed in this article.

Inhibitory effects of bevacizumab on corneal neovascularization and scarring in mice with herpes simplex keratitis / 中华实验眼科杂志

Background Corneal neovascularization and inflammation occur in herpes simplex keratitis (HSK).Aciclovir (ACV) is an antiviral medication which is primarily used for the treatment of HSV infection.Bevacizumab is an angiogenesis inhibitor which has the ability to slow the growth of corneal neovascularization.However,whether bevacizumab play treating effects on HSK is worth studing.Objective This study attempted to study the effects of bevacizumab on cornea lesion in mouse models of HSK.Methods The solution containing herpes simplex virus type-1 (HSV-1) of Mckrae strain was induced by cultured and infectious Vero cells and prepared by ten-times step dilution with free-serum DMEM,and plaque assay was used to detect the viral titers.HSV-1 of 1 ×l07 plaque-forming unit (PFU) in 0.6 μl was injected into the corneal stroma of 6 to 8-week-old SPF male C57BL/6 mice using a microliter syringe to establish latent HSK mouse models.The models were examined under the slit lamp microscope at day 5,7,11,14 and 17 after modeling as well as day 0,2,4 and 6 after recurrence,and the central cornea touch sensitivity was recorded.The models were divided into ACV-injected group,ACV+bevacizumabinjected group and normal saline-injected group,and 5 μl normal saline with 50 μg ACV,50 μg ACV + 5 μl bevacizumab or 10 μl normal saline was subconjunctivally iujected according to grouping in 4 eyes of each group,respectively.Twelve model eyes were exposed to ultraviolet (UV)-B to induce the recurrent HSK.Corneal wholemounts were prepared at day 9 after modeling for the assessment of corneal neovascularization and nerve fiber distribution by immunofluorescence assay of CD31 and β Ⅲ Tubulin antibodies.The areas of corneal neovascularization and scarring were mcasured with Image J software.The change rate of lesion was calculated and described as a ratio of lesion size at day 8 with day 0 after induction recurrence.Results The modeling success rate was over 80％,and all infected mice showed latent period at day 45 after modeling.Corneal opacification was the most serious at day 7 after modeling and day 2 after recurrence,and the largest corneal neovascular area was seen at day 15 after modeling and at day 2 after recurrence,and the central cornea touch sensitivity was the worst at day 9 after initial infection.The mean corneal lesion area was 3.348 mm2 in the ACV+bevacizumab-injected group,which was smaller than 3.930 mm2 in the ACV-injected group (Z=-2.309,P =0.021).The central corneal sensitivity in the ACV+bevacizumab-injected group was significantly higher than that in the normal saline-injected group (5.50± 0.71 versus 0.50± 1.41,Z =-2.397,P =0.029).The increase rate of corneal lesion area in the ACV +bevacizumabinjected group was evidently lower than that in the normal saline-injected group ([167.10 ± 52.53]％ versus [312.30± 74.18] ％,Z =-1.992,P =0.046).At the 7th day after modeling,the relative expressing levels of thymidine kinase (TK) and infected-cell protein-27 (ICP-27) mRNA in the corneal tissue and trigeminal ganglion were significantly increased at day 7 and reduced at day 45 after modeling,and the factors raised again at day 2 and retreated at day 7 after induction of recurrence.In addition,the expression of LAT mRNA peaked at day 45 after modeling and reduced gradually at day 2 after recurrence until a new increasing peak at day 7 after recurrence (all at P＜0.01).Immunofluorescence showed that compared with the normal saliue-injected group,the corneal new vessels were lessened and corneal never fibers were increased in the ACV-injected group and ACV +bevacizumab-injected group.Conclusions The combination of bevacizumab with ACV can inhibit corneal neovascularization and scarring in HSK mice,and bevacizumab exhibits a synergistic effect with ACV in management of HSK.

D-dimer can reflect severity of patients with community-acquired pneumonia and predict prognosis / 中华危重病急救医学

Objective To investigate the utility of plasma D-dimer and fibrinogen (FIB) for the severity assessments and predicting the prognosis of patients with community-acquired pneumonia (CAP).Methods The clinical data of patients with CAP admitted to First Affiliated Hospital of Fujian Medical University were retrospectively analyzed.The patients were divided into Ⅰ-Ⅴ level groups according to pneumonia severity index (PSI),and they were divided into non-survivors and survivors according to 30-day prognosis.The data including gender,age,PSI score,platelets count (PLT),white blood cell count (WBC),D-dimer,FIB,and C-reactive protein (CRP) were compared among groups.The correlations between PSI score and D-dimer,CRP as well as FIB were analyzed by Spearman or Pearson correlation analysis.Receiver operating characteristic curve (ROC) was used to assess the prognostic value of these indicators.Results A total of 499 patients with CAP were enrolled with 298 male and 201 female,the average age was (63.4 ± 17.8) years old,and the 30-day mortality was 6.4％ (32/499).There were 77,80,104,162 and 76 patients in PSI Ⅰ-Ⅴ level groups,and there were more male patients in PSI Ⅵ and Ⅴ level groups.There were no significant differences in PLT and FIB among the groups of different PSI levels,but the levels of WBC,D-dimer and CRP were significantly increased as PSI level increased from Ⅰ to Ⅴ (F1 =3.810,x 22 =102.361,F3 =7.070,all P ＜ 0.01).Compared with survivors,the non-survivors were elder (t =-4.773,P ＜ 0.001) with lower PLT (t =3.026,P =0.003)and higher WBC,PSI score,D-dimer and CRP levels (t1 =-2.545,t2 =-8.421,Z3 =-6.947,t4 =-3.770,all P ＜ 0.05).Plasma D-dimer levels in elderly patients (≥ 65 years old) were statistically higher than those in younger patients (＜ 65 years old;Z =-5.338,P ＜ 0.01).It was shown by correlation analysis that PSI score was positively correlated with D-dimer and CRP (r values were 0.475 and 0.260,both P ＜ 0.001),and no correlation was found between PSI score and FIB (r =-0.062,P =0.170).The area under the ROC curve (AUC) for predicting 30-day death of PSI score,D-dimer and CRP was 0.858 [95％ confidence interval (95％CI) =0.802-0.914],0.867 (95％CI =0.812-0.922) and 0.732 (95％CI =0.641-0.823).The combination of D-dimer and PSI score was better than any single indicator for predicting the prognosis with higher AUC up to 0.905 (95％CI =0.867-0.944),all P ＜ 0.001.The sensitivity and specificity for PSI in predicting 30-day death respectively were 78.1％ and 82.4％ with the cut-off of greater than 122,and those for D-dimer were 75.0％ and 82.9％ with the cut-off of greater than 2.10 mg/L,50.0％ and 84.4％ for CRP with the cut-off of greater than 100.50 mg/L.Conclusions D-dimer could well reflect the severity of CAP and be a good indicator for predicting the prognosis.The combination of D-dimer and PSI might improve the accuracy in predicting prognosis.

Death patterns of patients in Department of Respiratory Care Unit of General Hospi-tal of PLA / 军事医学

Objective To analyze retrospectively the death pattern, risk factors, and death time of 253 patients at the Respiratory Care Unit of General Hospital of PLA in order to improve care quality and reduce mortality.Methods The information of patients was extracted from the hospital information system ( HIS) , and then classified and calculated accord-ing to different time points.Results Between November and next March,the mortality rate was higher than in other months (P<0.05), accounting for 19.5%.Mortality of those admitted between 8∶01 and 9∶00 or between 23∶01 and 24∶00 was higher than at other times(P<0.05), accounting for 41.7%and 50.0%, respectively.There was statistically significant difference(P<0.01) in mortality between days of the week,with the highest on Saturday, accounting for 43.1%.Mortality on non-work days was higher than on workday(P<0.01), accounting for 38.3% and 13.2%, respectively.Mortality at off-hour was higher than at office time(8∶00-11∶30 and 14∶30-18∶00 on workday) (P<0.01), accounting for 31.3%and 5.2%, respectively.Logistic regression analysis showed that age, month of admission, and the hour of discharge were associated with the outcome.Conclusion The high mortality between November and next March may be related to the higher incidence of respiratory diseases in winter, air pollution and cold weather.High mortality is also significantly associ-ated with the care quality of the medical staff.