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Cryptococcus neoformans is an important pathogen causing opportunistic fungal meningitis. The pathogenic mechanism of cryptococcal meningitis remains unclear. We aimed to describe a practical approach for studying the pathologic features of cryptococcal central nervous system infection by immunofluorescence on paraffin-embedded brain of mice using different antigen retrieval methods. After 14 days of intratracheal inoculation of green fluorescent protein-tagged C. neoformans (H99-GFP), C57BL/6J mice brains were fixed in 4% paraformaldehyde and embedded in paraffin. Antigen retrieval methods such as microwaves, 1% sodium lauryl sulfate, 1 N HCl, pepsase, and tryptase were used on 5-µm paraffin sections and the effects were compared. The green fluorescence of H99-GFP persisted with antigen retrieval using 1% sodium lauryl sulfate. After immunofluorescent staining, H99-GFP, glial fibrillary acidic protein-tagged astrocytes, and ionized calcium-binding adapter molecule 1-tagged microglia could be observed clearly. Based on our results, we provide a practical approach for the further study of the interaction between C. neoformans and brain cells.
Subject(s)
Cryptococcus neoformans , Animals , Brain/metabolism , Cryptococcus neoformans/metabolism , Fluorescent Antibody Technique , Green Fluorescent Proteins/metabolism , Mice , Mice, Inbred C57BL , Paraffin Embedding , Staining and LabelingABSTRACT
In only a few months, the novel coronavirus severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused a global pandemic, leaving physicians, scientists, and public health officials racing to understand, treat, and contain this zoonotic disease. SARS-CoV-2 has made the leap from animals to humans, but little is known about variations in species susceptibility that could identify potential reservoir species, animal models, and the risk to pets, wildlife, and livestock. While there is evidence that certain species, such as cats, are susceptible, the vast majority of animal species, including those in close contact with humans, have unknown susceptibility. Hence, methods to predict their infection risk are urgently needed. SARS-CoV-2 spike protein binding to angiotensin converting enzyme 2 (ACE2) is critical for viral cell entry and infection. Here we identified key ACE2 residues that distinguish susceptible from resistant species using in-depth sequence and structural analyses of ACE2 and its binding to SARS-CoV-2. Our findings have important implications for identification of ACE2 and SARS-CoV-2 residues for therapeutic targeting and identification of animal species with increased susceptibility for infection on which to focus research and protection measures for environmental and public health.
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Objective To investigate the cryoprecipitate fracture hemorrhage clinical application value in the skull fracture hemorrhage,and provide reference for clinical treatment.Methods 80 patients with standard skull frac-ture hemorrhage were selected as the research subjects.The patients were randomly divided into observation group (40 cases)and the control group (40 cases).The control group was given hemostasis,dehydration,protecting stom-ach,antibacterial,nerve nutrition,reducing intracranial pressure,head up in bed and other symptomatic treatment.The observation group received the treatment besides in the control group and added the cryoprecipitate hemostasis,every time given 10U,every 6 -8h 1 time.According to the condition,they were given the cold sediment of 1 -3 times. Then,the cases that fit to give surgery we must cured them by the operation.After treatment 24h,the coagulation inde-xes were examined [prothrombin time (PT),activated partial thromboplastin time (APTT),thrombin time (TT),two D -dimer],plasma fibrinogen (Fbg),International Glasgow (GOS).Finally,we observed clinical prognosis in two groups.Results PT,APTT,TT were significantly shorter than before treatment in the observation group and the con-trol group(t =6.654,5.746,6.193 and 3.342,3.552,3.646,P <0.01 or P <0.05).The PT,APTT and TT of the observation group were significantly shorter than those in the control group (t =3.322,3.406,3.315,all P <0.05). Plasma Fbg was significantly higher than before treatment in two groups(t =5.762,3.592,P <0.01 or P <0.05). Fbg in the observation group was significantly higher than the control group(t =3.407,P <0.05).The clinical prog-nosis in the observation group was significantly better than the control group(χ2 =8.747,P <0.05).Conclusion Cryoprecipitate is a safe and efficient drug.It can effectively improve the blood coagulation dysfunction of patients with fracture of skull base,active endogenous coagulation system,improve hemostatic effect,and reduce the occurrence of progressive cerebral hemorrhage.Therefore,it can improve the prognosis of patients.
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Objective To observe the effect of metoclopramide combined with early enteral nutritional on patients with mechanical ventilation after craniocerebral operation.Methods A prospective randomized control study was conducted.56 patients with mechanical ventilation after craniocerebral operation were randomly divided into metoclopramide combined with early enteral nutritional group (observation group) and parenteral nutrition group (control group),28 in each group.The patients in the observation group received early enteral nutrition (24-48 h after operation),and intramuscular injection of metoclopramide (10mg Q8H) for 7 days.The patients in the control group received parenteral nutrition.The nutritional parameters,mechanical ventilation (MV) time,success rate of the ventilator weaning and the occurrence rate of pneumonia associated with ventilation (VAP) in one week,the incidence of adverse reaction were compared between the two groups.Results After 1 week therapy,the nutritioual parameters of the two groups showed no statistical significance(P ＞ 0.05);the observation group compared with the control group,3 、7days of blood glucose(P1 、P2) decreased significantly[(8.9 ± 1.6) mmol/L vs (11.4 ± 3.2) mmol/L,(8.2 ± 1.3) mmol/L vs (10.6 ± 2.1) mmol/L] (P1 =0.002,P2 =0.011),Mechanical ventilation (MV) time (P3) and the occurrence rate of VAP(P4) in one week also declined[(6.5 ±2.3)d vs (8.9 ±3.1)d,21.4％ vs 46.4％] (P3 =0.022,P4 =0.048),but within a week of success rate of the ventilator weaning improved (67.9％ vs 39.3％,P5 =0.032).Return rate (P6),gastrointestinal bleeding (P7) was significantly lower than those of the control group (17.9％ vs 42.9％,14.3％ vs 42.9％,P6 =0.042,P7 =0.038),but abdominal distention,diarrhea were no significant contrast (P ＞ 0.05).Conclusion Metoclopramide combined with early enteral nutritional may obviously improve the nutrition parameters,shorten the MV time,decrease the occurrence rate of VAP,not increase in side effects.
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binding,cellular organelle membrane,and cellular metabolic process of GO enrichment.For the KEGG pathways, the target genes mainly concentrated on the focal adhesion pathway.Conclusion There is a different expression of novel microRNA between SLE and NC groups.The target genes from differently-expressed novel microRNA may play an important role in the pathogenesis of SLE and clinical symptoms and may be the unique target for further research.
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Objective To explore the expression characteristics of vascular endothelial growth factor (VEGF) receptor (VEGFR).Methods The 123I-VEGF165 and 123I-VEGF121 were marked to human umbilical vein endothelial cell (HUVEC),several human tumor cell lines (HMC-1,A431,KU812,U937,HEP-1,HEP-G2,HEP-3B and Raji),a variety of human tumors and adjacent non-neoplastic tissues as well as peripheral blood cells.Then,the specific binding site maximal binding capacity (Bmax),dissociation constant (Kd) and concentration of 50％ required specific binding (IC5o) were analyzed.The affinity,quantity and specificity of different cells combined with 123I-VEGF165 and 123I-VEGF121 were judged.Results Two kinds of analogous 123I-VEGF165 binding sites on the surface of HUVEC were found.While,there was only one kind of 123I-VEGF121binding site.123I-VEGF121 was found on the special cell lines (HUVEC,HEP-1 and HMC-1) and special early tumors (early melanoma,ductal breast cancer,ovarian cancer and meningioma).Compared with peripheral blood cells and adjacent non-neoplastic tissues,the number of VEGFR of tumor cells was bigger.Among the 123I-VEGF165 marked VEGFR,the Bmax value of early melanoma,ductal breast cancer,hepatocellular carcinoma,papillary thyroid carcinoma,ovarian carcinoma,renal cell carcinoma were 45 ± 13,13 ± 3,25 ±8,5 ±2,42 ± 12,20 ±6,respectively.While among the 123I-VEGF121 marked VEGFR,the Bmax value of early melanoma,ductal breast cancer,ovarian carcinoma were 30 ± 8,8 ± 3,20 ± 6.123I-VEGF165 and 123I-VEGF121 had specific binding capacity with a variety of human tumor cells and tissues.Compared with 123I-VEGF121,there were more different kinds of tumor cells could be bound to 123I-VEGF165 with higher capacity.Conclusion 123I-VEGF165 may be a potential target of tumor imaging in vivo,and it is expected to be used to diagnose and treat tumors.
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Objective To investigate the epidemiological information of patients in pre-hospital medical care for our large and medium-sized cities and probe the patients' characteristic. Method The data in 2008 were exported from the computer databases of 8 large and medium-sized cities' emergency medical centers in our country.The thorough records of data were conducted to statistical analysis. Results ( 1 ) The scheduling time, running time, rescue time, returning time, total time and service radius in the pre-hospital medical care group were 2.16± 1.10(min), 14.01 ±6.82(min), 12.12±5.96(min), 14.08± 6.85(min), 42.34± 20.21(min)and 8.50±4.18(km), and the above parameter in the non-death group were 2.19 ± 1.13(min), 14.15 ± 7.14(min),11.60±6.72(min), 14.92 ±6.89(min), 41.86± 19.53(minutes) and 8.63±4.31(Km), and the above parameter in the death group were 2.10± 1.08(min), 13.68 ± 7.14(min), 25.25 ± 12.34(min), 13.75±6.48(min), 54.74 ± 25.47(min) and 7.86± 3.91(Km), and the above parameter in the non-sudden cardiac death group were2.09± 1.03(min), 13.58±6.78(min), 25.53± 12.34(min), 13.60± 6.54(min), 53.79±23.77(min) and 7.67 ± 3.86(Km), and the above parameter in the sudden cardiac death group were 2.12 ±1.02(min), 14.10±7.05(min), 24.79± 12.08(min), 13.79±6.61(min), 54. 80 ± 25. 36( min) and 7.90±3.92(Km) respectively. The scheduling time, running time, returning time and service radius in the death group were less than those of the non-death group, but the rescue time and total time of the former were more than those of the latter respectively ( P ＜ 0.05 or P ＜ 0. 001 ). The scheduling time and returning time didn' t have significant difference between the sudden cardiac death group and the non-sudden cardiac death group respectively ( P ＞ 0.05), but the running time, total time and service radius of the sudden cardiac death group were more than those of the non-sudden cardiac death group, and the rescue time of the former was less than that of the latter respectively ( P ＜ 0.05 or P ＜ 0.001 ). (2)The patients' amount in pre-hospital medical care group, the non-death group, the death group, the non-sudden cardiac death group and the sudden cardiac death group were at most in first quarter, and the least time slice of patients' amount were 4:00～ 6:00, 4:00～6:00, 4:00～ 6:00, 22:00～ 24:00, 2:00～4:00 respectively, and the most time slice of patients' amount were 20:00～ 22:00, 20:00～22:00, 8:00～ 10:00, 2:00 ～ 4:00, 8:00 ～ 10:00 respectively. (3)In 241 876 cases of pre-hospital medical care group, the patients' amount of trauma was at most, whose age grades was by far among21 ～50, and the others in sequence were nervous system, circulatory system, other group, digestive system, respiratory system and poisoning group respectively, whose age grades in nervous system, circulatory system and respiratory system was by far above 51, especially above 70. The patients' age grades in other group and digestive system had two climax age groups, which the one was 21 ～ 30, and the other was above 70. The patients' age grades in poisoning group was by far among 21 ～ 50, which the patients' amount of acute alcoholism was at the most. (4) In 12 568 cases of death group, the death amount of circulatory system, other group, respiratory system, nervous system and digestive system ranked at the lst,2nd,4th,5th 8th respectively, whose age grades was by far above 51, especially above 70,and the patients' amount of sudden cardiac death was at the most in the death amount of circulatory system. The death amount of trauma and poisoning group ranked at the 3rd, 6th respectively, whose age grades was by far among 21 ～ 50. (5)The total amount, the death amount and the sudden cardiac death amount of male patients were more than those of female patients. (6)The percentage of the death group to the pre-hospital medical care group was 5.20%, and the percentage of the sudden cardiac death group to the pre-hospital medical care group was 1.29%,and the percentage of the sudden cardiac death group to the death group was 24.87 %, and the percentage of the sudden cardiac death group to the circulatory system group was 67.33 %. Conclusions ( 1 )The trauma and the sudden cardiac death are the overriding reason of disease and the overriding reason of death in our large and medium-sized cities respectively. (2) It is very important to cut the death rate of the middle-old age patients by strengthening prevention and cure of cardiovascular and cerebrovascular diseases, discerning the critical illness early and improving the level of pre-hospital medical care. (3)It is a strong method to decrease the total amount and the death amount of the trauma, especially in traffic accident, by strengthening safety in production, observing traffic regulation and enhancing the legal awareness.
