Objective:To investigate the importance of cell block and immunohistochemistry in the accurate diagnosis of serous effusion.Methods:A retrospective study was conducted on 3 124 cases of serous effusion from the Department of Pathology, Beijing Hospital from 2018 to 2022, include 2 213 cases of pleural effusion, 768 cases of peritoneal effusion, 143 cases of pericardial effusion. There were 1 699 males (54.4%) and 1 425 females (45.6%), average age 69 years old. Of which 1 292 cases were prepared with cell blocks and examined with immunohistochemical stain.Results:The percentage of malignant diagnosis increased from 64.9% (839/1 292) to 84.0% (1 086/1 292) after cell block preparation, and 1 086 cases were accurately diagnosed with histological type and/or origin of primary tumor. The undetermined diagnosis of suspected malignancy decreased from 13.3% (172/1 292) to 0.1% (1/1 292) and that of atypical hyperplasia from 18.8% (243/1 292) to 0.4% (5/1 292). The negative result for malignancy rate increased from 3.0% (38/1 292) to 15.5% (200/1 292). The differences highlighted above were statistically significant (Pearson′s chi-squared test=12.739, P<0.01). Conclusion:Application of immunohistochemistry based on cell block can significantly improve malignant diagnosis in serous effusion, identify tumor origin and histological type as well as decrease the uncertain diagnosis.

Objective:To explore the relationship and underlying mechanism between exosomes derived from doxorubicin-resistant osteosarcoma cells and MDR1 and miRNAs. Methods:MG63 and U2OS cell lines were selected to construct doxorubicin-resistant strains, and the 50% inhibitory concentration (half maximal inhibitory concentration, IC 50) of drug-resistant and sensitive strains was detected by MTT, and fluorescence staining was performed at intervals of 15 min between 15 and 120 min to detect the change of fluorescence intensity. RT-PCR and Western Blot were used to detect the expression levels of MDR1 P-gp to verify the drug resistance of osteosarcoma cells. Exosomes were identified by particle size analysis and Western Bolt detection. The endocytosis of PKH26-labeled exosomes from doxorubicin-resistant cells was observed, and the proliferation level and migration of exosomes from doxorubicin-resistant cells co-cultured with osteosarcoma cells were detected by MTT assay and cell scratch assay. The differential expression levels of miRNAs in osteosarcoma-sensitive and drug-resistant cells were verified by sequencing and bioinformatics analysis and RT-PCR assay. Tumor growth, serum exosome identification and mRNA expression level of miR-21-5p in tumor-bearing nude mice between normal osteosarcoma cell group and drug-resistant group, drug-resistant+normal exosome group, drug-resistant+drug-resistant+drug-resistant exosome group were observed. MDR1 expression level in tumor tissue was detected by RT-PCR, Western Blot and immunohistochemistry. Results:The IC 50 of two adriamycin resistant strains were 2.21 vs. 11.81 μg/ml and 0.93 vs. 11.81 μg/ml, respectively, and the fluorescence intensity decreased faster than that of normal strains. The relative mRNA expression levels of MDR1 in two cell lines were normal 1.12±0.16, 1.02±0.11 and drug-resistant 2.15±0.10, 2.127±0.12, respectively. The relative protein expression of P-gp was normal 0.92±0.11, 0.73±0.10 and drug-resistant 0.46±0.03, 0.30±0.04, the differences were statistically significant ( P<0.05). Drug-resistant exosomes can enter osteosarcoma cells through endocytosis and concentrate in the cytoplasm when co-cultured with normal strains. Osteosarcoma cells were co-cultured with drug-resistant exosomes at 2, 4, 6, and 8 μg/ml adriamycin, respectively. Compared with normal group, the proliferation level in drug-resistant group was significantly increased. Compared with the normal cell group 35.95±3.92, 6.72±3.55 and the normal exosome group 51.22±5.55, 19.31±1.93, the drug-resistant cell group 54.20±9.32, 19.24±2.88 and drug-resistant exosome group 76.40±5.41, 30.26±4.87, all had significantly higher cell mobility, the difference was statistically significant ( P<0.05). Exosome sequencing and biogenic analysis of 10 highly upregated miRNAs to validate mRNA expression differences between normal and drug-resistant strains by RT-PCR, showing a significant increase in miR-21-5p expression level of drug-resistant strains (5.89±0.26 vs. 0.99±0.06; 1.05±0.07 vs. 8.80±0.93, P<0.05), the difference was statistically significant ( P<0.05). In MG63 and U2OS, the normal cell group and drug-resistant cell group, and the normal exosome group and drug-resistant exosome group were compared, the tumor volume and the terminal tumor weight of nude mice were increased to varying degrees. MRNA relative expression levels of miR-21-5p in serum exosomes of nude mice after drug intervention were 0.86±0.07 and 0.86±0.05 in normal cell group, respectively. The values were 1.13±0.12, 1.14±0.12 in drug-resistant cell group, 0.71±0.05, 0.75±0.03 in normal exosome group, and 0.90±0.07, 0.93±0.04 in drug-resistant exosome group. Compared with normal and drug-resistant strains, the expression levels of normal and drug-resistant exosome groups were increased, with statistical significance ( P<0.05). Conclusion:The exosomes of drug-resistant cells in osteosarcoma could enhance the proliferation level and migration ability of cells through intercellular transfer of MDR1 and miRNAs. The expression of MDR1 and miR-21-5p in drug-resistant cells and tumor-forming nude mouse serum and tumor tissues were up-regulated which suggested that it might be involved in regulating the drug resistance process of osteosarcoma.

Atrial fibrillation (AF) is the most prevalent cardiac arrhythmia seen in clinical settings, which has been associated with substantial rates of mortality and morbidity. However, clinically available drugs have limited efficacy and adverse effects. We aimed to investigate the mechanisms of action of andrographolide (Andr) with respect to AF. We used network pharmacology approaches to investigate the possible therapeutic effect of Andr. To define the role of Andr in AF, HL-1 cells were pro-treated with Andr for 1 h before rapid electronic stimulation (RES) and rabbits were pro-treated for 1 d before rapid atrial pacing (RAP). Apoptosis, myofibril degradation, oxidative stress, and inflammation were determined. RNA sequencing (RNA-seq) was performed to investigate the relevant mechanism. Andr treatment attenuated RAP-induced atrial electrophysiological changes, inflammation, oxidative damage, and apoptosis both in vivo and in vitro. RNA-seq indicated that oxidative phosphorylation played an important role. Transmission electron microscopy and adenosine triphosphate (ATP) content assay respectively validated the morphological and functional changes in mitochondria. The translocation of nuclear factor erythroid 2-related factor 2 (Nrf2) to the nucleus and the molecular docking suggested that Andr might exert a therapeutic effect by influencing the Keap1-Nrf2 complex. In conclusions, this study revealed that Andr is a potential preventive therapeutic drug toward AF via activating the translocation of Nrf2 to the nucleus and the upregulation of heme oxygenase-1 (HO-1) to promote mitochondrial bioenergetics.
Animals ; Rabbits ; Atrial Fibrillation/metabolism* ; Kelch-Like ECH-Associated Protein 1/metabolism* ; Signal Transduction ; NF-E2-Related Factor 2/pharmacology* ; Molecular Docking Simulation ; Oxidative Stress ; Energy Metabolism ; Mitochondria/metabolism* ; Inflammation/metabolism* ; Heme Oxygenase-1

With the development of social economy and the full opening of the three-child policy, more and more elderly people will become the main force in raising grandchildren in the future, and the mental health of the elderly in intergenerational raising has become the focus of the whole society. This article summarizes the causes of the intergenerational raising, the mental health status of the elderly in intergenerational raising, and reviews the influencing factors of the mental health of the elderly in intergenerational raising, in order to provide a reference for the construction of the social security system for the elderly and young people in China.

Objective: To understand the prevalence of overweight and obesity, dietary habits and main food intake frequency among primary school students in Shenyang, so as to provide a reference for exploring the effect of diet related factors on the development of overweight and obesity in children. Methods: A total of 2 041 students from two primary schools in a certain district of Shenyang were selected by convenience sampling in May 2017, with height and weight measured, meanwhile the questionnaire survey regarding dietary habits and the frequency of main food intake were administered. Results: The rates of overweight and obesity were 18.4% and 22.1% respectively, and the rate of overweight and obesity in boys (21.0%，27.8%) were significantly higher than that in girls (15.8%,16.2%)(χ 2=22.45,53.40，P<0.01). The results of univariate analysis showed that frequency of eating breakfast, eating speed, appetite, picky eaters or not, and the frequency of fruit, seafood and canned food intake were associated with overweight and obesity in primary school students (χ 2=7.67，97.92，229.70，95.88，6.40，6.58，7.96，P<0.05). Multivariate Logistic regression analysis showed that slow eating speed (OR=0.46, 95%CI=0.29-0.69) and normal eating speed (OR=0.47, 95%CI=0.32-0.69) were associated with lower rates of overweight and obesity; good appetite (OR=43.73, 95%CI=5.88-325.36) was associated with higher rates of overweight and obesity in primary school students (P<0.01). Conclusion The detection rate of overweight and obesity is relatively high among primary school students in Shenyang, and the rate of obesity is already higher than that of overweight; The frequency of common food intake has little impact on the development of overweight and obesity in primary school students, but fast eating speed and good appetite are the risks of overweight and obesity among them.

This article reported 2 cases primary renal Ewing sarcoma (PRES)/primitive neuroectodermal tumor (PNET). By reviewing literature, renal PRES/PNET has a high degree of malignancy, and early symptoms are not typical. It needs to be combined with clinical manifestations, imaging examinations and pathological examination results. At present, surgical treatment is the main treatment, combined with radiotherapy and chemotherapy or targeted treatment might help.

To access the efficacy of stents for spontaneous isolated dissection of the superior mesenteric artery (SIDSMA). The study is a prospective single-arm study which has been registered on Clinical Trials (NCT03916965). Clinical data and follow-up information of the SIDSMA patients who received stent implantation in the First Affiliated Hospital of Zhejiang University during April 1, 2019 and September 30, 2019 were collected. The patients were recommended to be followed up at 1, 3, 6 and 12 months. A total of 34 patients were enrolled. Their mean age was (54±8) years. Abdominal pain was the most common symptom. Patients received (2.1±0.6) stents on the average. Post-operation hospital stay was (2.7±1.6) days, and the patients were followed up for (2.3±1.9) months (CT angiography) and (5.5±1.7) months (clinical visit/phone call). There was no recurrence of abdominal pain. The CT angiography showed complete remodeling and incomplete remodeling took place in 23 and 9 patients (69.7% and 27.3%), respectively. Two patients (6.1%) had mild in-stent stenosis. No stent rupture or migration was reported. This study demonstrated a satisfactory short-term result of stents implantation for SIDSMA, which indicated the endovascular treatment could be the first-line therapy for SIDSMA.
Aneurysm, Dissecting ; Endovascular Procedures ; Humans ; Mesenteric Artery, Superior ; Middle Aged ; Prospective Studies ; Retrospective Studies ; Stents ; Treatment Outcome

Objective To explore the effect of lymphocytes on the innate immune cells in Rag1 and Rag2/IL2rγ gene knockout mice after hepatic ischemia and reperfusion injury (HIRI).Methods C57BL/6 mice (n =10),Rag1 knockout mice (n =10) and Rag2/IL2rγ knockout mice (n =10) were respectively divided into sham group and hepatic ischemia-reperfusion injury group by simple randomization,5 mice in each group.By using the model of hepatic ischemia-reperfusion injury in mice,changes of intrahepatic immune cells were detected by flow cytometry.Hepatic ischemia and reperfusion injury and changes of intrahepatic cell subsets were observed in immune system-deficient mice,both Rag1 and Rag2/IL2rγ knockout.Measurement data were expressed as ((x) ±s),and analyzed using independent samples t test.Results Flow cytometry results showed that immune cells,including NK cells,NKT cells,CD4+T cells,DNT cells,Kupffer cells,BMMs and neutrophils were increased after HIRI.Compared with sham group,Rag1 knockout mice displayed markedly increased proportion of Kupffer cells,BMMs and neutrophils after HIRI.And decreased serum ALT levels [from (1 776.25 ± 219.37) U/L to (932.33 ±58.77) U/L,P=0.003,t =7.350] and less hepatocellular necrosis were exhibited in Rag1 knockout mice after HIRI,comparing to C57BL/6 HIRI group.In addition,increased neutrophils were still observed in Rag2/IL2rγ knockout mice after HIRI,without increased proportion of Kupffer cells and BMMs.Compared with Rag1 knockout mice,ALT levels were further decreased from (932.33 ± 58.77) U/L to (309.00 ± 163.53) U/L (P=0.002,t =6.182) in Rag2/IL2rγ knockout mice.Conclusion Both Rag1 and Rag2/IL2rγ knockout mice exhibite less liver injury after HIRI comparing with C57BL/6 mice,indicating that T cells and NK cells aggravate the liver injury.Moreover,T cells do not affect the recruitment of Kupffer cells,BMMs and neutrophils,but regulate the recruitment of NK cells,while NK cells contribute to the activation of Kupffer cells and BMMs,but not neutrophil influx.

Objective To investigate the effect of intravenous tranexamic acid (TXA) on perioperative hidden blood loss in percutaneous pedicle screw fixation for thoracolumbar fractures.Methods A prospective study was conducted in the 113 patients who would be subjected to percutaneous pedicle screw fixation for thoracolumbar fracture from January 2017 to December 2017.They were randomly assigned into an observation group (n =58) receiving intravenous drip of 15 mg/kg TXA 30 minutes preoperation or a control group (n =55) receiving intravenous drip of normal saline solution 30 minutes preoperation.The total blood loss and hidden blood loss 24 hours postoperation,D-dimer volume,incidences of deep vein thrombosis and other complications were recorded and compared between the 2 groups.Results There were 54 patients in the observation group and 50 patients in the control group for statistic analysis.The observation group had significantly less total blood loss (319.0 ± 140.5 mL) and hidden blood loss (242.0 ± 143.4 mL) 24 hours postoperation than the control group (418.7 ± 188.1 mL and 354.7 ± 181.9 mL,respectively) (P ＜ 0.05).There were no significant differences between the 2 groups in operation time or intraoperative blood loss (P ＞ 0.05).The volume of postoperative D-dimer was significantly higher than the preoperative value in both groups (P ＜ 0.05).No thromboembolic events occurred in either group.Conclusion Intravenous TXA may significantly reduce intraoperative hidden blood loss with no increased rik of thromboembolic events in percutaneous pedicle screw fixation for thoracolumbar fractures.

Objective To compare minimally invasive percutaneous pedicle screw fixation and open pedicle screw fixation for neurologically intact thoracolumbar fractures.Methods A retrospective study was conducted in the 180 patients who had been treated for thoracolumbar fractures without neurological deficits from January 2016 to December 2016.Of them,93 were treated by minimally invasive percutaneous pedicle screw fixation and 87 by open pedicle screw fixation.The 2 groups were compared in terms of blood loss,radiological parameters,visual analogue scale (VAS) and Oswestry disability index (ODI).Results Compared with the open surgery group,the minimally invasive surgery group had significantly shorter operating time (95.8 ±33.4 min versus 106.3 ±30.9 min),significantly less intraoperative blood loss (65.8 ±40.3 mL versus 183.1 ± 77.5 mL),significantly less total blood loss in theory 24 hours after surgery (374.7 ± 160.6 mL versus 614.8 ± 242.6 mL) and significantly shorter hospital stay (5.2 ± 2.0 d versus 6.7 ± 2.7 d),but significantly longer C-arm exposure time (23.6 ±4.2 min versus 12.4 ±4.1 min) and significantly more hidden blood loss 24 hours after surgery (308.9 ± 159.0 mL versus 243.5 ± 195.5 mL) (P ＜ 0.05).Compared with preoperation,significant improvements were observed at one week postoperation and the last follow-up in the 2 groups regarding the percentage of anterior height of the fractured vertebral body and cobb angle (P ＜ 0.05),but there were no significant differences in the percentage of anterior height of the fractured vertebral body or cobb angle between the 2 groups at one week postoperation or at the last follow-up (P ＞ 0.05).At 3 days postoperation,significant better pain relief was observed in the minimally invasive surgery group than in the open surgery group (P ＜ 0.01),but at the last follow-up no obvious pain was reported in either group.At the last follow-up,there was no significant difference between the 2 groups in ODI (6.2 ± 1.1 versus 6.0 ± 1.4) (P =0.320).Conclusions In the treatment of neurologically intact thoracolumbar fractures,minimally invasive percutaneous pedicle screw fixation may lead to shorter operating time,less blood loss and shorter hospital stay but no poorer radiological outcomes or long-term patient-reported outcomes than the open pedicle screw fixation.However,it should be noted that the former may lead to a higher volume of hidden blood loss.