ObjectiveTo explore the application effects of environmentally friendly reagents versus traditional reagents in tissue processing， paraffin sectioning， hematoxylin-eosin （HE） staining， special staining， and immunohistochemistry. MethodsPancreatic， renal， splenic， hepatic， pulmonary， skeletal muscle， and cardiac tissues were collected from mice and New Zealand rabbits， and randomly divided into a traditional group （Group A）， an environmentally friendly reagent I group （Group B）， and an environmentally friendly reagent Ⅱ group （Group C）. The processed tissue sections from all three groups were subjected to paraffin sections， HE staining， phosphotungstic acid-hematoxylin staining， and Mallory trichrome staining. The pancreatic tissue sections were subjected to immunohistochemical detection of insulin protein. Evaluation criteria included tissue morphological integrity， embedding operability， section quality， and staining effects. ResultsNo significant differences were observed among the three groups of samples in terms of fixation efficacy， embedding operations， and section quality. After HE staining， all groups exhibited bright nuclear and cytoplasmic staining with clear contrast， with excellent-and-good rates of 98%， 99%， and 97% for Groups A， B， and C， respectively， showing no statistically significant differences （A vs. B， P=0.561； A vs. C， P=0.651）. In phosphotungstic acid-hematoxylin staining， Groups A and C showed clear structural contrast and bright staining for striated muscle and collagen fibers， whereas Group B exhibited uneven staining. The excellent-and-good rates were 100%， 10%， and 95% for Groups A， B， and C， respectively， with significant differences between Group A and Group B （P＜0.01） but no statistical difference between Group A and Group C （P=0.311）. In Mallory trichrome staining， Groups A and B outperformed Group C. The excellent-and-good rates were 95%， 90%， and 15% for Groups A， B， and C， respectively， with significant differences between Group A and Group C （P＜0.01）， but no statistical difference between Group A and Group B （P=0.548）. Immunohistochemical results showed that Group A had a clean background with no non-specific staining， while Groups B and C exhibited significant non-specific staining， with statistically significant differences in non-specific staining rates between the groups （A vs. B， 5% vs. 100%， P＜0.01； A vs. C， 5% vs.100%， P＜0.01）. ConclusionIn HE staining， environmentally friendly reagents demonstrated effective tissue fixation and staining results comparable to traditional reagents， meeting the teaching requirements. For special staining， Group B and Group C performed better in Mallory trichrome staining and phosphotungstic acid-hematoxylin staining， respectively. However， environmentally friendly reagents exhibited non-specific staining issues in immunohistochemistry， where further optimization is required.

Objective To investigate the expression and significance of 25-hydroxyvitamin D[25(OH)D],immune cell subsets,and helper T cell(Th)1 and Th2 in patients with unexplained recurrent spontaneous abortion(URSA).Methods A total of 307 URSA patients were enrolled as URSA group,which was further divided into non-maintenance of pregnancy group(n=127)and ma-intenance of pregnancy group(n=180)based on pregnancy maintenance status.Another 307 healthy pregnant women in the same period were selected as control group.The levels of 25(OH)D,immune cell subsets,and Th1/Th2 expression were compared among these groups.Pearson correlation analy-sis was used to explore the correlations of 25(OH)D and immune cell subsets with Th1/Th2 expres-sion in URSA patients.Results Compared with the control group,the URSA group had significantly lower levels of 25(OH)D and interleukin-10(IL-10),and higher levels of CD3+,CD3+CD4+,CD3+CD8+,CD4+/CD8+,CD16+CD56+,CD19+,interleukin-2(IL-2),and tumor necrosis fac-tor-α(TNF-α)(P＜0.001).Compared with the maintenance of pregnancy group,the non-maintenance of pregnancy group had significantly lower levels of 25(OH)D and IL-10,and higher levels of CD3+,CD3+CD4+,CD3+CD8+,CD4+/CD8+,CD16+CD56+,CD19+,IL-2,and TNF-α(P＜0.001).Pearson analysis revealed that 25(OH)D in URSA patients was significantly negatively correlated with IL-2 and TNF-α(P＜0.05),while positively correlated with IL-10(P＜0.05).Immune cell sub-sets,including CD3+,CD3+CD4+,CD3+CD8+,CD4+/CD8+,CD16+CD56+,and CD19+were significantly positively correlated with IL-2 and TNF-α(P＜0.05),while negatively correlated with IL-10 in URSA patients(P＜0.05).Conclusion There are abnormalities in 25(OH)D,im-mune cell subsets,and Th1/Th2 cytokine expression in URSA patients,and these expressions differ in URSA patients with different pregnancy outcomes.The underlying mechanism may be related to the regulation of Th1/Th2 cytokine balance.

Objective To explore the potential prognostic factors of patients with acute ischemic stroke(AIS)after undergoing endovascular mechanical thrombectomy and to construct an effective predictive model.Methods A retrospective analysis of clinical data was conducted on 202 patients with anterior circulation large vessel occlusion AIS from 2 stroke centers.All patients received endovascular mechanical thrombectomy treatment,with treatment and follow-up lasting at least 90 d.Basic demographic characteristics,medical records,and baseline blood biomarker data were collected,and the potential prognostic indicators for AIS after 90 d were screened using least absolute shrinkage and selection operator(LASSO)-logistic regression analysis.Results It was found that alcohol drinking(P=0.029),hypertension(P=0.001),diabetes mellitus(P=0.021),stroke or transient ischemic attack(P=0.049),systolic blood pressure on admission(P=0.009),diastolic blood pressure on admission(P=0.038),blood glucose(P=0.003),white blood cell count(P=0.001),neutrophil count(P=0.001),fibrinogen(P=0.010),systemic immune-inflammation index(P=0.008)and neutrophil-to-lymphocyte ratio(NLR)(P＜0.001)were associated with adverse clinical outcomes.Nine significant prognostic determinants were screened through LASSO-logistic regression analysis.Multivariate logistic regression analysis revealed that male sex(P=0.008),smoking history(P=0.013),hypertension(P=0.011),lymphocyte(P=0.028),fibrinogen(P=0.016),and NLR(P＜0.001)were significant predictive factors for poor prognosis in AIS patients after endovascular thrombectomy treatment.The constructed prognostic model had an accuracy of 76.2％,a sensitivity of 78.2％,a specificity of 71.7％,and a positive predictive value of 86.7％.Conclusion The predictive model established in this study can assist clinicians in identifying high-risk patients with AIS who have undergone endovascular thrombectomy,and it provide guidance for formulating individualized treatment strategies.

Objective To analyze the outcomes of catheter ablation for persistent atrial fibrillation(AF)and the independent risk factors for its recurrence in the elderly.Methods A total of 194 patients with persistent AF who underwent catheter ablation at our department from January 2019 to December 2021 were enrolled in this study.They were divided into elderly group(≥60 years old,99 cases)and non-elderly group(＜60 years old,95 cases).Their surgical characteris-tics,postoperative complications and recurrence were compared between the two groups,and the independent risk factors for postoperative recurrence were analyzed in the elderly group.Results Advanced age,higher B-type natriuretic peptide,larger proportions of hypertension and coronary heart disease,and increased CHA2DS2-VASc and HAS-BLED scores,while lower male ratio and estimated glomerular filtration rate were observed in the elderly group than the non-elderly group(P＜0.05,P＜0.01).The elderly group had a higher proportion of left atrial fibrosis than the non-elderly group(30.3％vs 8.4％,P=0.001).Postoperative complications in the elderly group in-cluded 1 case of pericardial effusion and 2 cases of hematoma at the puncture site,and all of these were improved after treatment.There were no significant differences in the 1-year success rate(71.7％vs 69.5％,P=0.763)or recurrence rate during blanking period(21.2％vs 21.1％,P=0.981)between the elderly and non-elderly groups.AF duration(HR=1.020,95％CI:1.007-1.032,P=0.002)and recurrence during blanking period(HR=6.781,95％CI:3.078-14.935,P=0.001)were independent risk factors for postoperative recurrence in the elderly group.Conclu-sion Catheter ablation is safe and effective in the treatment of persistent AF in the elderly.The elderly patients with long duration of AF and recurrences during blanking period are more likely to experience recurrences within 1 year after ablation.

Objective, accurate and noninvasive assessment of the condition and prognosis of traumatic brain injury (TBI) patients in the early stage is the basis of TBI treatment. Glasgow coma scale (GCS) is the most common clinical method for assessing trauma condition. However, when patients are sedated, intubated or have language or movement disorders, it is often impossible to make an accurate assessment by GCS. In addition, GCS cannot provide direct evidences of pathological damage in brain tissues. The CT scoring system has emerged in recent years and is a non-invasive evaluation system based on CT images. The evaluation process can objectively reflect the severity of brain tissue damage without being interfered by sedation or intubation. The authors review the progress of researches on CT scoring systems to assess the prognosis of patients with craniocerebral injury, so as to provide a reference for clinicians to choose appropriate injury assessment methods and make clinical decisions.

Objective To explore the effects of cranioplasty onneurological functionin patients based on the cerebral CT perfusion technique. Methods Twenty cases of patients receiving cranioplasty were rerecorded during the study period,and they wererespectively scanned by CT perfusion within 72 hours before and 2 weeks after the cranioplasty. Meanwhile,the neurological function was evaluated by neurological function scale. Results The difference of cerebral blood flow before and after cranioplasty was statistically significant(P<0.05), whereas the difference of cerebral blood volume,transit time to the peak and mean transit time was not statistically significant(P>0.05).Correlation analysis showed that the preoperativedifference ratio of thecerebral blood infusio-nis not correlated with the neurological function score(P > 0.05). The changes of preoperative and postoperative difference rateof the cerebral blood infusionwas correlated with the functional independence measure(P < 0.05), whereas not with mini-mental state examination(P > 0.05). Conclusions The neurological function of the patients after cranioplasty may be improved.This improvement may benefit from the improvement of cerebral blood flow after cranioplasty.

Objective To determine the intervention of mycophenolate mofetil (MMF) in glial scar formation and learning and memory function in a rat model of diffuse axonal injury (DAI).Methods Ninety-six SD rats were randomly divided into sham group, normal saline (NS) group and mycophenolate mofetil (MMF) group according to the random number table, with 32 rats per group.Immunohistochemistry was used to detect activated microglia cells, activated astrocytes and chondroitin sulphate proteoglycanns (CSPGs) in the hippocampus.Inage-Pro Plus software was used to quantitatively assess the changes of activated microglia cells, activated astrocytes and CSPGs.Morris water maze was applied for testing rat learning and memory function.Integrated absorbance (IA) of major constituents (microglia, astrosyte, chondroitin sulphate proteoglycan) of the glial scar was determined and analyzed for the correlation with the parameters of MWM.Results At 7, 14 and 28 days after injury, MMF group showed decreased IA of activated microglia in the hippocampus compared to sham and NS groups (P ＜ 0.05).At 7-11 days after injury, percent distance and percent time in the target quadrant of Morris water maze did not differ significantly among the three groups and were not related to the IA of glial scar.At 28-32 days after injury, percent distance and percent time in the target quadrant of Morris water maze lowered significantly in MMF group.At 28 days after injury, IA of the glial scar had a positive correlation with mean speed and mean escape latency, but negative correlation with percent distance and time in the target quadrant that measured in Morris water maze at 28-32 days after injury.Conclusion MMF significantly attenuates glial scar formation into the hippocampus and improves learning and memory function in rats during the recovery stage when administered in the early stage after DAI.

Objective To investigate the role of interleukin-17 (IL-17) in spinal dorsal horns in neuropathic pain (NP) in rats and its effect on activation of astrocytes.Methods In vivo experiment Sixty-four male SPF Sprague-Dawley rats,aged 6-8 weeks,weighing 180-200 g,were randomly divided into 3 groups using a random number table:control group (group C,n =16),sham operation group (group S,n =24) and group NP (n =24).The animals were anesthetized with intraperitoneal pentobarbital sodium,the L5,6 spinal nerves of the left side of the rat were gently separated and exposed,tightly ligated with 5-0 silk suture and transected.In group S,the L5,6 spinal nerves of the left side of the rat were only exposed.In group C,no operation was performed.Mechanical pain threshold was measured at day 1 before operation and days 1,3,5,7,10 and 14 after operation.The expression of IL-17,IL-6,IL-1β and tumor necrosis factor-alpha (TNF-α) mRNA in the spinal dorsal horn was determined using quantitative real-time PCR at day 7 and day 14 after operation.At day 7 after operation,the activation of astrocytes in the spinal dorsal horn was detected.In vitro experiment Primarily cultured astrocytes of neonatal rats were randomly divided into 4 groups using a random number table:control group (group C,n=22),10 ng/ml IL-17 group (I10 group,n=18),50 ng/ml IL-17 group (I50 group,n-18) and 100 ng/ml IL-17 group (I100 group,n=22).In I10,I50 and I100 groups,the astrocytes were incubated with the culture medium containing 10,50 and 100 ng/ml IL-17,respectively.The proliferation of astrocytes was detected by MTT at 24,48 and 72 h of incutation or culture.The expression of IL-6,IL-1β and TNF-α mRNA was determined using quantitative real-time PCR.Results In vivo experiment Compared with group C,the mechanical pain threshold was significantly decreased at 3-14 days after operation,the expression of IL-17,IL-6 and IL-1β mRNA in the spinal dorsal horn was up-regualted at 7 days after operation,and the activation of astrocytes was increased in group NP,and no significant change was detected in the mechanical pain threshold at each time point after operation in group S.In vitro experiment Compared with group C,the proliferation of astrocytes was significantly increased at 48 h of incubation in I10 and I50 groups,the proliferation of astrocytes was significantly increased at 48 and 72 h of incubation,and the expression of IL-6 and IL-1β mRNA was up-regulated in I100 group,and no significant change was found in the proliferation of astrocytes in group S.Conclusion Up-regulated expression of IL-17 in spinal dorsal horns may be involved in the maintenance of NP,and the mechanism is related to promoted activation of astrocytes and induced inflammatory responses in rats.

Objective: To explore the guiding significance of transpulmonary thermodilution pulse-indicated continuous cardiac output (PiCCO) in treatment for patients with acute inferior wall myocardial infarction (AIMI). Methods: A total of 26 AIMI patients in intensive care unit of cardiology department in our hospital from Jul 2012 to Jan 2014 received PiCCO and ultrasonic cardiography (UCG) to monitor cardiac output (CO) and cardiac index (CI), and their correlation analysis. Results: When PiCCO placement and after placement 72h， PiCCO monitoring CI were （2.77±0.77）L•min-1•m-2, (3.17±0.39) L•min-1•m-2 respectively, there was significant difference（P＜0.01）, UCG measured CI were (2.49±0.64）L•min-1•m-2, (2.63±0.24) L•min-1•m-2, there was no significant difference（P＞0.05）; PiCCO monitoring CO were（4.78±1.06）L/min, (5.08±1.53) L/min respectively, there was significant difference（P＜0.05）, UCG measured CO were（4.51± 0.86）L/min, (4.57±0.91) L/min, there was no significant difference（P＞0.05）; and CI，CO measured by PiCCO were significantly higher than those of UCG group (P＜0.01 both) after PiCCO placement 72h. Conclusion: Pulse-indicated continuous cardiac output can offer more sensitive hemodynamic indexes compared with UCG, which possesses important treatment guiding significance in patients with acute inferior wall myocardial infarction and unstable hemodynamics.

Objective To establish a stable orthotopic transplantation nude mouse model of the human pancreatic cancer and to explore the role of monitoring tumor growth with noninvesive MRI.Methods The tumors cells suspension made by the subcutaneous injection of human pancreatic cancer cell lines PANC1 were used as the source of tissue for orthotopic implantation of tissue.and transplanted into the pancreas of 20 BALB/C-nu nude mice.After implantation,the successful rate,tumor formation time,tumor growth speed,tumor shape and the change of signal of the tumor were monitored and recorded noninvasively by MRI.At the end of the 7th week,all the specimens were examined by pathological methods.Results Thirty-five percent (7/20)mice with implantation of primary human PANC1 adenocarcinoma cells were detected to have orthotopic implanted tumors by MRI after 15 days,and all the 20 nude mice developed pancreatic tumor within 27 days after operations,and the successful rate was 100%.Compared with adjacent normal tissue,the T1 WI imaging of 90%(18/20)of all the tumors showed uniformly hypo-intense signal,10%(2/20)showed iso-intense signal,the T2W 75%(15/20)showed uniformly hyper-intense signal.The tumor size 2,3,4,5,6,7 weeks after implantation was(912.6±2.4)mm3,(94.3±11.2)mm3,(175.9±82.5)mm3,(395.8±126.6)mm3,(1290.2±167.2)mm3,(1583.4±87.4)mm3,respectively.Pathologic examination confirmed poody differentiated pancreatic adenocatcinoma and it remained the primary tumor's biolosic features.Conclusions The orthotopic transplantation nude mouse model was in accordance with the feature of human pancreatic cancer and was easy for noninvasive monitoring,which provided an effective and stable in vivo experimental system.