Objective To study dectin-1 mRNA expression in lung of different immune status mice infected with Aspergillusfumigatus, and to explore the influence of immunosuppressive agent on the expression of dectin-1 and its relationship with the progression of disease. Methods Mice were divided into four groups which were normal control, immuncompromised, immuncompromised with A.fumigatus inoculation and immuncompetent with A.fumigatus inoculation groups. We explored the kinetic mRNA expression of dectin-1 in lung of different groups by real-time quantitative PCR. Pulmonary fungal burden assessment was performed to reflect the progressing of disease during the experimental time course. Results On day 3 after inoculation, pulmonary fungal burden of the immuncompromised mice was higher than that of the immuncompetent group. On day 1 and 3, dectin-I mRNA expression in lung of the immuncompromised group was much lower than that of the normal control. On day 3, dectin-1 mRNA expression in lung of the immuncompetent mice infected with A.fumigatus was much higher than that of the immuncompromised with A.fumigatus inoculation and the normal control groups ( P < 0.05 ). Conclusion During the infection, expression of dectin-1 in lung of the immuncompetent group was strikingly increased, which may play an important role on the defence to A.fumigatus invasion. Cyclophosphamide inhibited the expression of dectin-1 in lung of mice which may be one of the mechanisms of the invasive pulmonary aspergillosis development induced by eyclophosphamide.

Objective:To observe the effect of attenuated-dose aflibercept in the treatment of retinopathy of prematurity(ROP).Methods:A non-randomized controlled study was conducted, and 76 eyes of 38 ROP pediatric patients treated in First Affiliated Hospital of Zhengzhou University from December 2018 to May 2020 were enrolled.According to the requirements of their guardians, the patients were divided into ranibizumab group with 42 eyes of 21 cases and attenuated-dose aflibercept group with 34 eyes of 17 cases, and received intravitreal injection of ranibizumab 0.025 ml (0.25 mg) or aflibercept 0.012 5 ml (0.5 mg) according to grouping respectively.Retcam fundus photography was used to observe the treatment response at 1 week, 2, 4 weeks and 2, 3, and 6 months after treatment, and the effective rate at the end of follow-up was calculated.The intraocular pressure was measured with Icare PRO magnetic rebound tonometer at 1 minute, 10, and 30 minutes after injection. The ocular and systemic complications were observed during the 6-month follow-up period.All the guardians signed the informed consent prior to treatment.This study was approved by the Ethics Committee of the First Affiliated Hospital of Zhengzhou University (No.2020-KY-228).Results:The effective rates of single ranibizumab and attenuated-dose aflibercept were 90.5% (38/42) and 88.2% (30/34), respectively, with no significant difference between the two groups ( χ2=0.10, P=0.75). The intraocular pressure of the ranibizumab group at 1 minute and 10 minutes after the operation were higher than those of the attenuated-dose aflibercept group, and the difference was statistically significant (both at P<0.01). The intraocular pressure recovered to the baseline level at 30 minutes after the operation.In the ranibizumab group, 4 eyes were ineffective after a single injection, among which 2 eyes were effective after second intravitreal injection of ranibizumab and 1 eye was effective after retinal laser photocoagulation treatment and 1 eye underwent vitrectomy due to the progress of retinal detachment one week after intravitreal injection, and the posterior retina reattached well.In the attenuated-dose aflibercept group, 4 eyes did not respond to treatment, of which 3 eyes were effective after second intravitreal injection of aflibercept, and 1 eye was effective after retinal laser photocoagulation.No ocular or systemic complications were observed during the followed-up period. Conclusions:Reduced dose of aflibercept is safe and effective in the treatment of ROP, and has little influence on intraocular pressure.

Aim To investigate the protective effect of allicin against EA. hy926 endothelial cell injury in-duced by PM2. 5 and the possible mechanism. Meth-ods The samples of fine particulate matter ( PM2. 5 ) were collected and made into suspension. Different concentrations of PM2. 5 ( 20 , 200 , 400 mg · L-1 ) were added to EA. hy926 cell. The viability and apop-tosis of EA. hy926 cell, the protein levels of p-ERK1/2, Bax and Bcl-2 in the EA. hy926 cell, the contents of tumor necrosis factor-α( TNF-α) , interleukin-6 ( IL-6 ) , and malonaldehyde ( MDA ) , the activities of su-peroxide dismutase ( SOD ) and lactic dehydrogenase ( LDH) in the EA. hy926 cell culture supernatant were measured by MTT assay, flow cytometry, Western blot, enzyme-linked immunosorbent assay ( ELISA ) and colorimetry, respectively. Allicin at different con-centrations(5,20,40 mg·L-1 ) or a specific inhibitor of ERK1/2 signaling pathway PD98059 ( 20 μmol · L-1 ) was added into the EA. hy926 cell to observe the effect of allicin. Results Compared with control group, PM2. 5 significantly increased the apoptosis, the contents of TNF-α, IL-6 and MDA, the activity of LDH, the protein levels of p-ERK1/2 and Bax/Bcl-2 ratio, but decreased the viability and SOD activity in the EA. hy926 cell(P<0. 05). Compared with PM2. 5 group, allicin significantly decreased the apoptosis, the contents of TNF-α, IL-6 and MDA, the activity of LDH, the protein levels of p-ERK1/2 and Bax/Bcl-2 ratio, but increased the viability and SOD activity in the EA. hy926 cell ( P <0. 05 ) . Conclusion Allicin displays a significant protective effect against EA. hy926 endothelial cell injury induced by PM2 . 5 and its mechanism may be related to the attenuations of in-flammation and oxidative stress via the inhibition of ERK1/2 pathway.

AIM:To investigate the effect and the mechanism of tanshinone ⅡA in attenuating PM2.5-induced human umbilical vein endothelial EA.hy926 cell injury.METHODS:The samples of fine particulate matter (PM2.5) were collected in Guangzhou and made into suspension.Different concentrations (0, 20, 200 and 400 mg/L) of PM2.5 were added to EA.hy926 cells.The viability and apoptosis of EA.hy926 cells, the protein levels of p-p38 MAPK, Bax and Bcl-2 in the EA.hy926 cells, the contents of interleukin-6 (IL-6), tumor necrosis factor-α(TNF-α) and malonaldehyde ( MDA) , and the activity of superoxide dismutase ( SOD) and lactic dehydrogenase ( LDH) in the EA.hy926 cell culture supernatant were measured by MTT assay, flow cytometry, Western blot, ELISA and colorimetry, respectively.Tanshinone ⅡA at different concentrations (5, 10 and 20 μmol/L) or a specific inhibitor of p38 MAPK pathway, SB203580 (20μmol/L) , was added into the EA.hy926 cells to observe the effect of tanshinone ⅡA.RESULTS:Compared with control group, PM2.5 significantly increased the apoptosis, the contents of IL-6, TNF-αand MDA, the activity of LDH, and the protein levels of p-p38 MAPK and Bax/Bcl-2 ratio, but decreased the viability and SOD activity in the EA.hy926 cells (P<0.05).Compared with PM2.5 group, tanshinone IIA significantly decreased the apoptosis, the contents of IL-6, TNF-αand MDA, the activity of LDH, and the protein levels of p-38 MAPK and Bax/Bcl-2 ratio, but increased the viabil-ity and SOD activity in the EA.hy926 cells (P<0.05).CONCLUSION:Tanshinone ⅡA attenuates PM2.5-induced EA. hy926 cell injury via the inhibition of p38 MAPK pathway.

OBJECTIVE:To study the inhibitory effects of berberine on EA.hy926 human umbilical vein endothelial cells (EA. hy926 cells) injury induced by particulates with no more than 2.5 μm air aerodynamic diameter in atmospheric (PM2.5),and its p38 mitogen-activated protein kinase(MAPK)signal pathway mechanism. METHODS:PM2.5 samples were collected and hatched EA.hy926 cells with concentrations of 0(blank control),20,200 and 400 mg/L for 24 h. The survival rate and apoptosis rate of cells,contents of IL-6,TNF-α and MDA,activities of SOD and LDH,protein levels of p-p38 MAPK,Bcl-2 and Bcl-2 associated X protein (Bax) were detected. The above indexes of EA.hy926 cells in blank control group,PM2.5 group (200 mg/L PM2.5), p38 MAPK pathway-specific blocker SB203580 group (20 μmol/L SB203580+200 mg/L PM2.5),berberine low-,medium- and high-concentrations groups(5,10,20 μmol/L berberine+200 mg/L PM2.5)were also determined. RESULTS:Compared with blank control,survival rate of cells,SOD activity and Bcl protein decreased after 200,400 mg/L PM2.5 hatched;apoptosis rate of cells, contents of IL-6,TNF-α and MDA,LDH activity,protein levels of p-p38 MAPK and Bax increased (P<0.05),in concentra-tion-dependent manner. Compared with PM2.5 group,survival rate of cells,SOD activity and Bcl-2 protein increased in berberine medium-,high-concentrations groups and SB203580 group;apoptosis rate of cells,contents of IL-6,TNF-α and MDA,LDH ac-tivity,protein levels of p-p38 MAPK and Bax decreased (P<0.05). CONCLUSIONS:Berberine attenuates PM2.5-induced EA. hy926 cells injury via the inhibition of p38 MAPK pathway.

Objective To evaluate the performance of VITEK 2-Compact GN13 methods for testing imipenem susceptibility of Klebsiella pneumoniae and assess the reliability of its Advanced Expert System (AES).Methods A retrospective study was conducted with a total of 157K. pneumoniae strains, which were isolated from blood and intra-abdominal infections in the First Affiliated Hospital of Nanchang University from 2014 to 2015. Thein vitro minimum inhibitory concentration (MIC) values of imipenem were determined by disc diffusion, VITEK 2-Compact GN13 and broth microdilution methods, respectively. Categorical agreement (CA) rates of disc diffusion and VITEK 2-Compact GN13 methods were determined using broth microdilution as reference method. The genes encoding ESBLs and carbapenemase were screened by PCR and sequencing analysis. The phenotypic confirmatory tests such as modified Hodge test, PCR and DNA sequencing were used to confirm the resistance mechanism and evaluate the reliability of AES in interpreting the imipenem susceptibility of K. pneumoniae.Results Among the 157 isolates, 64 and 8 were identified as resistant and intermediate strains by broth microdilution method, respectively; 52 and 10 were tested as resistant and intermediate strains by disc diffusion method, respectively; 54 and 13 were determined as resistant and intermediate strains by VITEK 2-Compact GN13 method, respectively, while 70 and 3 were judged as resistant and intermediate strains by VITEK 2-Compact GN13 method plus AES validation. The CA of disc diffusion and VITEK 2-Compact GN13 methods compared with broth microdilution method were all higher than 90 %. However, the major error (ME) rate was 3.8 % and very major error (VME) rates were all 0.6 % in imipenem susceptibility testing by VITEK 2-Compact GN13 and disc diffusion. The imipenem susceptibility of 16 strains were modified by the AES, which eliminated 0.6 % VME, but increased major error by 1.3 % and minor error by 1.9 %. Phenotypic confirmatory tests showed that 75 % (12/16) of these strains were validated as producers of both ESBLs and carbapenemase, which was consistent with the result of AES validation. PCR and DNA sequencing analysis proved that 62.5 % (10/16) of these strains produce IMP-4/KPC-2 /NDM-1 and ESBLs.Conclusions Both disc diffusion and VITEK 2-Compact GN13 methods can be used for testing the imipenem susceptibility of K. pneumoniae isolates with reliable and accurate results. Attention should be paid to the possibility of ME and VME when testing imipenem susceptibility. The VME can be avoided by the AES mechanism. However, AES intervention will increase ME and minor error, which may be associated with decreased expression of carbapenemase.

Objective By studying clinical features, treatment and prognosis of eosinophilic gastroenteritis of infants resulted from milk protein allergy an, to improve the diagnosis and treatment level of eosinophilic gastroenteritis. Methods 24 cases of infants which diagnosed eosinophilic gastroenteritis were chosen from June of 2010 to January of 2014 in children’s Hospital of XX province and By retrospective analysis clinical manifestations, endoscopic features, histopathology, treatment and prognosis of the 24 cases. Results The 24 cases who were vomiting, paroxysmal crying, abdominal distension (100.00%), which accompanied by haematemesis 23 cases (95.83%), 1case (4.17%) hematochezia, 17cases (70.83%) eczema, 21 cases (87.50%) mild to moderate anemia, 1 cases (4.17%) severe anemia, 19 cases (79.17%) the increasing of peripheral blood eosinophil cells, 8 cases (33.33%)the increasing of IgE of the serum and 4 cases (16.67%) the test of antibody of the Helicobacter pylori in Serum was positive; 3 cases (12.50%) were milk protein allergy by the detecting of food allergen-speciifc IgE antibodies, the endoscopic characteristics were hyperemia, edema, erosion, ulcer of gastric and duodenal mucosa. Among them, 24 cases (100.00%) were gastritis, 5 cases (20.83%) duodenitis and 1 cases (4.17%) duodenal ulcer. The histopathology of the 24 cases revealed that there were gastric or duodenal eosinophils infiltration (> 20/HPF) and were all associated with mast cell infiltration; By antisecretory, protection of the gastrointestinal mucosa and the obviating of milk protein had a satisfactory treatment effect, 24 cases of children with oral general formula milk test conifrmed that the milk protein allergy, The 3 cases of the patients were reviewed by 8~12 weeks after gastroscope, and the mucosa of the duodenum was smooth, Eosinophils were/HPF<8, mast cells were/HPF<5. Conclusion There are no speciifc clinical and endoscopic manifestations in eosinophilic gastroenteritis of infants resulted from milk protein allergy, gastrointestinal mucosa eosinophil inifltration and simultaneously are accompanied by abnormal mast cell inifltration;Mucosal type without the use of corticosteroids, through milk protein avoidance treatment can achieve satisfactory results, But definite diagnosis must rely on biopsy and eosinophils, combined with avoidance stimulation test of milk protein can further confirmed, But the excitation test should be at least 10 days of observation of children, and carefully recorded symptoms, so as not to delay the missed diagnosis of CMPA.

Objective To investigate the changes of serum cytokines and explore their role in infants with cytomegalovirus (CMV) infection. Methods We recruited 41 positive CMV-IgM plus normal ALT infants (other disease group), 30 positive CMV-IgM plus abnormal ALT infants (hepatitis group) and 30 healthy infants (control group) in the study. The levels of tumor necrosis factor-?(TNF-?), interferon-?(IFN-?) and interleukin-4 (IL-4) in serum were measured with ELISA. The association between TNF-? and ALT was analyzed. Results The levels of TNF-?, IFN-?, IL-4 were higher, and IFN-?/IL-4 was significantly lower in the two CMV infection groups than the control group. Compared with other disease group, the changes of TNF-?, IL-4, and IFN-?/IL-4 showed significance in hepatitis group. The level of TNF-? showed a positive association with ALT in hepatitis group(r=0.76,P

The effects of oxygen partial pressure on cryopreservation of the cells with organ preservation solution were explored. Hypoxic UW solution was made by purging the UW solution with argon. The pig proximal tubule epithelial cells (LLC-PK1 cells) were cryopreserved in hypoxic UW solution (Ar-UW group) or standard UW solution (UW group) at 4 degrees C for 48 h. Trypan blue staining and LDH detection were performed to evaluate the injury of the cells. The results showed that the oxygen partial pressure in Ar-UW group was significantly declined from 242+/-6 mmHg to 83+/-10 mmHg. After cryopreservation at 4 degrees C for 48 h, LDH leakage rate and Trypan blue-stained rate in Ar-UW group were (11.3+/-3.4)% and (10.5+/-4.7)%, respectively, which were significantly lower than in UW group [(49.5+/-6.9)% and (47.6+/-9.3)% respectively, both P<0.01]. It was concluded that lower oxygen partial pressure of UW solution was more beneficial to the cryopreservation of LLC.
Adenosine ; Allopurinol ; Cell Hypoxia ; Cell Line ; Cryopreservation ; Cryoprotective Agents ; Epithelial Cells/*cytology ; Glutathione ; Insulin ; Kidney Tubules, Proximal/cytology ; Organ Preservation Solutions ; Oxygen/pharmacology ; Raffinose ; Swine ; Tissue Preservation/methods

In order to explore the method to prepare hypoxia UW solution and the stability and preservation of hypoxia UW solution, UW solution was purged by argon or air for 15 min or 60 at a flow rate of 0.8 or 2 L/min, and the oxygen partial pressure of UW solution was detected. The hypoxia UW solution was exposed to the air or sealed up to preserve by using different methods, and the changes of oxygen partial pressure was tested. The results showed that oxygen partial presure of 50 mL UW solution, purged by argon for 15 min at a flow rate of 2 L/min, was declined from 242+/-6 mmHg to 83+/-10 mmHg. After exposure to the air, oxygen partial pressure of hypoxia UW solution was gradually increased to 160+/-7 mmHg at 48 h. After sealed up by the centrifuge tube and plastic bad filled with argon, oxygen partial pressure of hypoxia UW solution was stable, about 88+/-13 mmHg at 72 h. It was concluded that oxygen of UW solution could be purged by argon efficiently. Sealed up by the centrifuge tube and plastic bag filled with argon, oxygen partial pressure of UW solution could be stabilized.
Adenosine/chemical synthesis ; Allopurinol/chemical synthesis ; Anoxia ; Glutathione/chemical synthesis ; Insulin/chemical synthesis ; Organ Preservation/*methods ; Organ Preservation Solutions/*chemical synthesis ; Oxygen/*analysis ; Partial Pressure ; Raffinose/chemical synthesis