Objective Effective tissue perfusion fixation plays an important role in immunofluorescence experiments and other studies .The aim of this study was to modify the method for perfusion fixation of rat brain tissues and improve the efficiency of the procedure . Methods Conventional thoracotomy was modified by cutting the skin from the xiphoid along the mediventral line to the lower jaw to expose both sides of the thorax and then freeing the chest wall .The tip of the infusion needle was cut off and the needle in-serted into the left ventricle , pushed to the ascending aorta , and fixed there to prevent sliding .Perfusion and fixation began after tho-racic aortic occlusion.The regulatory valve was opened , followed by infusion of 0.9%physiological saline and cutting the right atrial appendage .When the perfusate became clear , the infusion tube was replaced , without removal of the needle , and fixed with 4%paraformaldehyde .The fixation process was slowed down gradually till the neck became stiff , which indicated its completion . Results The head, neck and upper limbs were all fixed desirably by this method , which shortened the operation time , saved 4%paraformalde-hyde, and achieved satisfactory immunofluorescence results as compared with conventional procedures . Conclusion The modified method is simple , obviously improves the results of perfusion fixation of rat brain tissue , and therefore deserves to be popularized .

Objective To monitor the expression patterns of bcr-abl in chronic myeloid leukemia (CML) patients during treatment with imatinib mesylate and evaluate the detection of MRD by RQ-PCR method. Methods The ABI Prism 7500 Sequence Detection System using Taqman fluorogenic probes was used to quantify target gene. bcr-abl mRNA was detected by RQ-PCR in 106 CML patients. The normalized quotient (NQ) of bcr-abl mRNA was calculated as followings: NQ=bcr-abl mRNA copy numbers/abl mRNA copy numbers. Results The NQ of BCR-ABL mRNA was well correlated with the progression of disease and the number of Ph+ cell (r =0.9824 and 0.9346, respectively). The NQ was decreased rapidly in 62 patients and kept in low level for a long time, and only 2 of them were relapsed. For 8 patients, after treatment the NQwere decreased initially and increased sharply, 7 of them were relapsed after 5-9 months. After treatment the NQ of 31 patients were still>0.1, 11 patients were relapsed after a short remission and 7 were ineffective or progression. Out of 5 patients whose NQ were fluctuated and had little regularity, but all of them had a continuing remission. Conclusion RQ-PCR is a more sensitive technique in the detection of bcr-abl fusion gene.It is an important method to monitor the tumor cell during the treatment with imatinib mesylate in CML patients.

Objective To investigate the predictive value of Alberta stroke program early CT score on diffusion-w eighted imaging (DWI-ASPECTS) for predicting new cerebral microbleeds (CMBs) in patients w ith acute middle cerebral artery infarction. Methods The patients w ith acute middle cerebra artery infarction w ere enroled prospectively. MRI examinations w ere completed w ithin 48 h on admission and they w ere examined again at 10 to 14 d after onset. Susceptibility-w eighted imaging (SWI) w as use to detect
CMBs. DWI-ASPECTS w as used to assess the infarction extent. Results A total of 82 patients w ith acute middle cerebra artery infarction w ere enroled, including 27 females and 55 females. Their ages w ere 71.7 ± 8.9 years. Eighteen patients (22.0%) had old CMBs, 25 (30.5%) had new CMBs, 57 (69.5%) did not have new CMBs. Compared w ith the non-new CMB group, DWI-SPECTS (3.20 ±1.73 vs.7.11 ±1.69;t = 9.573, P <0.001) w as low er, NIHSS scores (16.20 ±4.06 vs.12.63 ±5.06; t = 3.111, P = 0.003) w ere higher, there w ere more patients w ith atrial fibrilation ( 40.0% vs.15.8%; χ2 = 5.722, P = 0.017), proportion of intensive antiplatelet therapy ( 0% vs.28.1%; P = 0.002) w as low er, there w ere more large artery atherosclerosis type ( 60.0% vs.29.8%; χ2 = 6.650, P = 0.010 ), more cardiogenic cerebral embolism type (36.0% vs.5.3%; P = 0.001), and less smal artery occlusion type ( 0% vs.57.9%; P <0.001) in the new CMB group, and there w ere no statistical differences in the other indexes. Multivariate logistic regression analysis show ed that after adjusting age, sex, alcohol, histories of hypertension, hyperlipidemia, diabetes, atrial fibrilation and previous stroke or transient ischemic attack history, the higher the DWI-ASPECT scores ( > 5), the risk of new CMBs w ould decrease 86 % (odds ratio 0.14, 95%confidence interval 0.17 -0.48; P < 0.001). Receiver operating characteristic curve analysis show ed that the sensitivity of prediction of DWI-ASPECTS ≤5 for the new CMBs w as 87.7%, specificity w as 88.3%, and the area under the curve w as 0.940. Conclusions DWI-ASPECTS can effectively predict the new CMBs in patients w ith acute middle cerebra artery infarction.

Objective: To observe the effect of edaravone combining ulinastatin on brain protection in patients of type A aortic dissection (AAD) after total arch replacement. Methods: A total of 60 AAD patients with total arch replacement in our hospital from 2014-09 to 2016-01 were prospectively studied. Based on peri-operative application of edaravone and ulinastatin, the patients were divided into 2 groups: EU group: 1) the patients received ulinastatin 300000 U/8h and edaravone 0.5mg/Kg/12h from administration to 3 days post-operation, 2) during cardiopulmonary bypass, the patients received ulinastatin 300000 U/2h and edaravone 0.5mg/Kg; Control group, the patients had no such treatment.n=30 in each group. The following items were observed:①operative condition;②blood levels of speciifc brain injury markers as S-100 and neuron speciifc enolase (NSE) at different time points: beginning of surgery (T0), opening aorta clamp (T1), right after cardiopulmonary bypass (T2), entering ICU (T3), 24h post-operation (T4) and 3 days post-operation (T5); ③post-operative condition. Results:①Durations of operation, cardiopulmonary bypass, cardiac arrest and bilateral antegrade selective cerebral perfusion (BACP), the frequency of BACP and UACP (unilateral antegrade selective cerebral perfusion), the lowest rectal temperature and blood levels of S-100, NSE at T0 were similar between 2 groups.②Compared with Control group, EU group had decreased S-100 and NSE from T1 to T5,P<0.05.③The in-hospital and ventilation time, frequency of PND and TND, the patients with CSS score≥16 before discharge and the in-hospital death rate were similar between 2 groups,P>0.05. Conclusion: Edaravone combining ulinastatin had brain protective effect in AAD patients after total arch replacement;it may reduce blood speciifc brain injury markers while the clinical signiifcance should be further investigated.

Objective To investigate the lipid profiles of patients with primary Sjogren's syndrome (pSS) and to analyze the correlation between abnormal serum lipids and the inflammationsof SS. Methods One hundred and fourteen pSS patients and 20 gendermatehed healthy controls were studied. Serum lipids were measured in both groups. Results There was statistically significant difference between SS and healthy controls, and the serum HDL-c and apoA<,1 concentrations were significantly lower in patients (P<0.05). The incidence of abnormal serum lipids was 39.5% in these patients. Patients with abnormal lipids had longer course of disease, higher ESR level, lower salivary flow rate and more frequent parotid gland enlargement than those without abnormal lipids(P<0.05). Logistic regression analysis revealed statistically significant association between serum lipids levels and occurrence of parotid gland enlargement. Conclusion Findings from this study suggest that patients with SS have altered lipid profiles and the decrease of apoA, and HDL-c levels may be the correlated factors of SS. The inflammation of SS may cause changes in lipids metabolisms.

Background Intracorneal macrophages play a critical role in corneal neovascularization (CNV)by secreting relative chemokines.But macrophages are characteristic by heterogeneity which has different biologic functions under different induction or stimulation from microenvironment.Objective This study was to detect the effects of chemokine (C-X3-C motif) ligand 1 (CX3CL1) and chemokine (C-C motif) ligand 2 (CCL2) on macrophages in vitro.Methods CNV was induced by corneal alkali burn in the left eyes of 20 male BALB/c mice aged 7-8 weeks.The CNV was evaluated under the slit lamp microscope 4 days after alkali burn,and then the corneal sections were prepared after mice were sacrificed.The expressions of CCR2 and CX3CR1 in the corneal specimens were detected by histo-fluorescence staining.Human peripheral blood mononuclear cells were separated using density gradient centrifugation and incubated in RPMI-1640 medium containing 10％ fetal bovine seruml(FBS) with 30 μg/L granulocyte-macrophage colony-stimulating factor (GM-CSF).The cells were divided into CD68 +CCR2 group and CD68+CX3CR1 group,and the percentage of the CX3CR1 and CCR2 expressions in the infiltrated macrophages of corneal specimens and human monocyte-derived macrophages was assayed by flow cytometry.The cultured cells were stimulated using human recombinant CX3CL1 and CCL2 proteins,and real-time PCR was used to detect the relative expressions of angiogenesis-related factors in macrophages.Results CNV was found in corneas 4 days after alkali burn and the CNV onsets from corneal limbus to central zone observed by a slit lamp.CCR2 and CX3CR1 were expressed in the F4/80-positive macrophages in alikali burned corneas.The macrophages grew for two weeks and appeared more dead cells in without GM-CSF group,but in GM-CSF induced group,the number of macrophages was increased.The percentage of CX3CR1-positive cells was 75％ and that of CCR2-positive cells was 45％.Real-time PCR showed that expression level of vascular endothelial growth factor (VEGF) mRNA increased and that ADAMTS-1 mRNA or TSP-1 mRNA decreased on macrophages after CCL2 stimulation,with significant differences in the 150 mg/L CCL2 group compared with the control group (t =-5.09,P =0.03 ; t =3.01,P =0.04 ; t =4.27,P =0.02).However,the VEGF mRNA expression decreased and ADAMTS-1 mRNA and TSP-1 mRNA increased after CX3CL1 stimulation,showing significant differences between the 150 mg/L CX3CL1 group and the control group (t=6.35,P=0.O2;t=-2.92,P=0.04; t=-3.81,P=0.03).Conclusions These results suggest that the macrophages have high heterogeneity.CCL2-and CX3CL1-expressing macrophages can regulate the expressions of angiogenesis-related factors.Macrophage chemokine signal may be a good target for treatment of neovascular ocular disease.

Objective:To observe the clinical features of cytomegalovirus (CMV) retinitis (CMVR)- related uveitis after hematopoietic stem cell transplantation (HSCT).Methods:A retrospective clinical study. From October 2015 to May 2020, 14 cases of 21 eyes of CMVR patients with CMVR after HSCT confirmed by the ophthalmological examination of The First Affiliated Hospital of Soochow University were included in the study. Among them, there were 5 males with 8 eyes and 9 females with 13 eyes. The average age was 35.12± 12.24 years old. All the affected eyes were examined by slit lamp microscope combined with front lens and fundus color photography. At the same time, fluorescein fundus angiography (FFA) was performed to examine 10 eyes of 5 cases; 3 cases of 3 eyes were examined for inflammatory cytokines in aqueous humor. All eyes received intravitreal injection of ganciclovir; patients with a history of systemic CMV infection received intravenous infusion of ganciclovir/foscarnet. The retinal lesions in the eye were completely resolved or the aqueous CMV-DNA was negative as a cure for CMVR. The uveitis symptoms, signs, FFA manifestations and the test results of inflammatory factors in aqueous humor before and after the CMVR cure was observed. The follow-up time after CMVR was cured was 3-42 months, and the average follow-up time was 14.28±13.12 months.Results:All eyes with CMVR were diagnosed with retrocorneal dust and/or stellate keratic precipitates (KP), anterior chamber flare and cells, and varying degrees of vitreous flocculent opacity; the retina was typical of a mixture of hemorrhage and yellow-white necrosis like "scrambled eggs with tomatoes". After CMVR was cured, there were 16 eyes (71.4%, 10/14) in 10 cases with KP, anterior chamber flare, cell and vitreous opacity. FFA examination revealed that the majority of retinal leakage during the active period of CMVR was necrotic foci and surrounding tissues; after CMVR was cured, the majority of retinal leakage was the retina and blood vessels in the non-necrotic area. The test results of inflammatory factors in aqueous humor showed that interleukin (IL)-6, IL-8, and vascular endothelial cell adhesion molecules were significantly increased in the active phase of CMVR; after 3 months of CMVR cured, inflammatory factors did not increase significantly.Conclusion:CMVR-associated uveitis after HSCT show as chronic panuveitis, with no obvious eye congestion, KP, anterior chamber flare, cell and vitreous opacity, and retinal vessel leakage which could exist for a long time (>3 months).

Objective To understand the usage of national medical insurance negotiated drugs(hereinafter referred to as"negotiated drugs")at Tianjin Second People's Hospital and to provide references for optimizing and adjusting the hospital's drug catalog.Methods A retrospective study was conducted on the usage of negotiated drugs from January 1,2018 to December 31,2022 to compare changes in the unit price of drugs,the quantity and amount of sales,the usage frequency(DDDs)and the daily average cost(DDC),etc before and after the negotiation.Results Between 2018 and 2022,the varieties of negotiated drugs used in the hospital increased from the original 5 to 24.Among the 13 drugs analyzed for comparison,the unit prices of 11 drugs had been reduced after negotiation,and 7 drugs were included in the medical insurance and outpatient-specific disease payment directory.The average decrease in DDC was 36.43%,and the average increase in DDDs was 1 770.31%.The implementation of this policy had increased the accessibility of medication for patients and significantly increased sales quantity.Conclusion The quantity of sales of negotiated drugs significantly increased through reducing the unit price of drugs and including them in the scope of medical insurance payment,etc.These increase the pharmacoeconomic viability of negotiated drugs,effectively reduce the burden on patients,promote rational drug use in hospitals,and improve the access and efficiency of drugs.

Objective:To analyze the prevalence of allergic rhinitis in Chinese children from 2001 to 2021, in order to provide data support for the prevention and treatment of allergic rhinitis in children.Methods:"Allergic rhinitis" "children" "adolescent" "infant" "prevalence" "epidemiology" were used in the main search terms. The combination of Mesh words and free words was adopted. CNKI, CBM, VIP, WanFang Data, PubMed, Web of Science, Embase and The Cochrane Library for publications between January 1, 2001 and December 31, 2021 were searched systemically and data were extracted from eligible studies by two independent reviewers. Supplementary collection was made by identifying retrospective references from the included literature. After study quality assessment, Meta analysis was completed using Stata 16.0 software.Results:A total of 20 cross-sectional studies were included, involving 54 886 cases. Meta analysis results showed that the overall prevalence of allergic rhinitis among the participants was 18.46% (95% CI:14.34%-22.59%). Subgroup analysis showed that the prevalence of allergic rhinitis from 2012 to 2021 (19.75%) was higher than that from 2001 to 2011 (14.81%), and the difference was statistically significant ( P<0.001). The prevalence of different regions from high to low was East China (22.77%), North China (20.82%), Northwest China (17.77%), Central China (16.62%), Southwest China (16.33%), Northeast China (16.16%) and South China (7.29%) respectively, the difference was statistically significant ( P<0.001). The prevalence of male (20.73%) was higher than that of female (16.34%), and the difference was statistically significant ( P<0.001). The prevalence of Han nationality(17.31%) was higher than that of ethnic minorities (15.93%), and the difference was statistically significant ( P<0.001). Conclusion:The prevalence of allergic rhinitis in Chinese children is high and the prevalence in children varies by publication year, region, sex and nationality.

Objective:To analyze the prevalence of allergic rhinitis in Chinese children from 2001 to 2021, in order to provide data support for the prevention and treatment of allergic rhinitis in children.Methods:"Allergic rhinitis" "children" "adolescent" "infant" "prevalence" "epidemiology" were used in the main search terms. The combination of Mesh words and free words was adopted. CNKI, CBM, VIP, WanFang Data, PubMed, Web of Science, Embase and The Cochrane Library for publications between January 1, 2001 and December 31, 2021 were searched systemically and data were extracted from eligible studies by two independent reviewers. Supplementary collection was made by identifying retrospective references from the included literature. After study quality assessment, Meta analysis was completed using Stata 16.0 software.Results:A total of 20 cross-sectional studies were included, involving 54 886 cases. Meta analysis results showed that the overall prevalence of allergic rhinitis among the participants was 18.46% (95% CI:14.34%-22.59%). Subgroup analysis showed that the prevalence of allergic rhinitis from 2012 to 2021 (19.75%) was higher than that from 2001 to 2011 (14.81%), and the difference was statistically significant ( P<0.001). The prevalence of different regions from high to low was East China (22.77%), North China (20.82%), Northwest China (17.77%), Central China (16.62%), Southwest China (16.33%), Northeast China (16.16%) and South China (7.29%) respectively, the difference was statistically significant ( P<0.001). The prevalence of male (20.73%) was higher than that of female (16.34%), and the difference was statistically significant ( P<0.001). The prevalence of Han nationality(17.31%) was higher than that of ethnic minorities (15.93%), and the difference was statistically significant ( P<0.001). Conclusion:The prevalence of allergic rhinitis in Chinese children is high and the prevalence in children varies by publication year, region, sex and nationality.