BACKGROUND:Previous studies have successfully constructed erythropoietin-overexpressed umbilical cord mesenchymal stem cells.It was found that the apoptosis of ischemic and hypoxic human neuroblastoma cell line(SH-SY5Y)was significantly reduced by erythropoietin-overexpressed umbilical cord mesenchymal stem cells. OBJECTIVE:To explore the possible neuroprotective mechanisms of erythropoietin-overexpressed umbilical cord mesenchymal stem cells against ischemic-hypoxic SH-SY5Y and their associated epigenetic mechanisms. METHODS:Oxygen-glucose deprivation was applied to ischemia-hypoxia-induced SH-SY5Y cell injury,and multifactorial assays were applied to detect the expression levels of inflammatory factors in the cells before and after hypoxia and co-culture,respectively,with mesenchymal stem cells,as well as lentiviral-transfected null-loaded plasmids of the negative control mesenchymal stem cells and erythropoietin-overexpressed umbilical cord mesenchymal stem cells.The expression levels of supernatant inflammatory factors were detected by multifactor assay after co-culture.Proteomics was used to detect the differentially expressed proteins of negative control mesenchymal stem cells and erythropoietin-overexpressed umbilical cord mesenchymal stem cells.Cleavage under targets and tagmentation sequencing was applied to detect genomic H3K4me2 modification,and joint analysis was conducted with RNA-sequencing.Lentiviral vector infection was applied to construct the stable knockdown of REST in SH-SY5Y cells.qRT-PCR and western blot assay were performed to detect the expression level of REST.The apoptosis was detected by flow cytometry after co-culture of oxygen-glucose deprivation treatment with erythropoietin-overexpressed umbilical cord mesenchymal stem cells.The expression difference of H3K36me3 group proteins was detected by western blot assay,and transcriptome sequencing was performed to analyze the differentially expressed genes. RESULTS AND CONCLUSION:(1)Compared with the control group,monocyte chemotactic protein 1,interleukin-6,interleukin-18,and interleukin-1 beta,interferon α2,and interleukin-23 levels significantly increased in the cerebrospinal fluid supernatant of patients with ischemic-hypoxic encephalopathy(P<0.01).(2)After co-culturing SH-SY5Y cells with erythropoietin-overexpressed umbilical cord mesenchymal stem cells under ischemia and hypoxia,the expression levels of monocyte chemotactic protein 1 and interleukin-6 were significantly reduced.(3)Analysis of protein network interactions revealed significant downregulation of monocyte chemotactic protein 1,interleukin-6 related regulatory proteins CXCL1 and BGN.(4)Transcriptome sequencing analysis found that pro-inflammatory genes were down-regulated,and functional enrichment of histone modifications,and the expression of transcription factors REST and TET3 significantly up-regulated in the erythropoietin-overexpressed umbilical cord mesenchymal stem cell group compared with the negative control mesenchymal stem cell group.(5)Combined analysis of transcriptome sequencing and cleavage under targets and tagmentation revealed changes in epigenetic levels as well as significant activation of the promoter regions of transcription factors REST and TET3.(6)Stable knockdown REST in SH-SY5Y cells was successfully constructed;the transcript levels of REST mRNA and protein expression were both decreased.(7)After the REST knockdown SH-SY5Y cells were co-cultured with erythropoietin-overexpressed umbilical cord mesenchymal stem cells,apoptosis was significantly increased and H3K36me3 expression was significantly decreased.Transcriptome sequencing results showed that the expression of inflammation-related genes Aldh1l2 and Cth,as well as apoptosis-suppressor genes Mapk8ip1 and Sod2 was reduced at mRNA transcription level(P<0.01).(8)It is concluded that erythropoietin-overexpressed umbilical cord mesenchymal stem cells activated the expression of REST and TET3 by altering the kurtosis of H3K4me2 and upregulated the modification level of H3K36me3,which in turn regulated the expression of inflammation-related genes Aldh1l2 and Cth,as well as apoptosis-suppressor genes Mapk8ip1 and Sod2,and facilitated neuronal survival.

Objective To evaluate the impact of optimizing the stroke green channel on the efficiency of acute ischemic stroke management in a county hospital. Methods A retrospective analysis of the emergency stroke green channel treatment data from Sixian People’s Hospital from May 2020 to April 2021 (before optimization of the green channel) and from May 2021 to April 2022 (after optimization of the green channel) was conducted. The rates of intravenous thrombolysis (IVT) and mechanical thrombectomy (MT) patients, as well as door-to-needle time (DNT), door-to-puncture time (DPT), and the modified Rankin scale (mRS) scores of patients three months post-treatment before and after the optimization of the stroke green channel were compared. Results Within one year before and after optimization of the green channel, the number of acute visits for ischemic stroke was 3 143 and 2 623, respectively. Before optimization, 84 and 51 underwent IVT and MT, respectively. After optimization of the green channel, the ratios of patients underwent IVT (n＝215) and MT (n＝103) significantly increased, and both DNT and DPT were significantly shortened (P＜0.000 1); the proportion of MT patients with an mRS score of 0-2 at 3 months post-discharge significantly increased (46/99 vs 13/46, P＝0.038). Conclusion After optimizing the green channel at Sixian People’s Hospital, the efficiency of stroke treatment has significantly improved, and the patients’ prognosis improved.

Objective:To further improve the understanding of paroxysmal nocturnal hemoglobinuria (PNH), we retrospectively analyzed and summarized the clinical characteristics, treatment status, and survival status of patients with PNH in Zhejiang Province.Methods:This study included 289 patients with PNH who visited 20 hospitals in Zhejiang Province. Their clinical characteristics, comorbidity, laboratory test results, and medications were analyzed and summarized.Results:Among the 289 patients with PNH, 148 males and 141 females, with a median onset age of 45 (16-87) years and a peak onset age of 20-49 years (57.8% ). The median lactic dehydrogenase (LDH) level was 1 142 (604-1 925) U/L. Classified by type, 70.9% (166/234) were classical, 24.4% (57/234) were PNH/bone marrow failure (BMF), and 4.7% (11/234) were subclinical. The main clinical manifestations included fatigue or weakness (80.8%, 235/289), dizziness (73.4%, 212/289), darkened urine color (66.2%, 179/272), and jaundice (46.2%, 126/270). Common comorbidities were hemoglobinuria (58.7% ), renal dysfunction (17.6% ), and thrombosis (15.0% ). Moreover, 82.3% of the patients received glucocorticoid therapy, 70.9% required blood transfusion, 30.7% used immunosuppressive agents, 13.8% received anticoagulant therapy, and 6.3% received allogeneic hematopoietic stem cell transplantation. The 10-year overall survival (OS) rate was 84.4% (95% CI 78.0% -91.3% ) . Conclusion:Patients with PNH are more common in young and middle-aged people, with a similar incidence rate between men and women. Common clinical manifestations include fatigue, hemoglobinuria, jaundice, renal dysfunction, and recurrent thrombosis. The 10-year OS of this group is similar to reports from other centers in China.

Objective:To investigate the role of miR-765 in papillary thyroid carcinoma (PTC) cells and further uncover the associated signaling mechanism.Methods:qPCR was used to assess miR-765 expression in normal human thyroid cell line (Nthy-ori 3-1) and human PTC cell lines (B-CPAP and TPC-1). PTC cells were divided into blank control group (BC) without special treatment, negative control group (NC) that was transfected with negative control sequences, and miR-mimic group that was transfected with miR-mimic. Transfection with miR-mimic was used to up-regulate the expression of miR-765 in PTC cells. CCK-8, plate colony formation, wound-healing, and Transwell invasion assays were used to assess the proliferation, migration, and invasion of PTC cells, respectively. Western blot assay was used to assess the level of nuclear β-catenin, the key protein of the Wnt/β-catenin pathway, in PTC cells.Results:The level of miR-765 expression of PTC cells was significantly lower than that of Nthy-ori 3-1 cells (B-CPAP, P=0.0003; TPC-1, P=0.0003). Transfection with miR-mimic significantly up-regulated miR-765 expression in PTC cells (B-CPAP, P<0.0001; TPC-1, P<0.0001). Results of CCK-8 assay (B-CPAP, P<0.05; TPC-1, P<0.05), plate colony formation assay (B-CPAP, P=0.0001; TPC-1, P<0.0001), wound-healing assay, and Transwell invasion assay (B-CPAP, P=0.001; TPC-1, P=0.0014) showed that up-regulating the expression of miR-765 significantly inhibited the proliferation, migration, and invasion of PTC cells. Western blot results showed that up-regulating the expression of miR-765 significantly reduced nuclear β-catenin (B-CPAP, P=0.0039; TPC-1, P=0.0004) . Conclusion:up-regulating the expression of miR-765 inhibits the proliferation, migration, and invasion of PTC cells and the Wnt/β-catenin signaling pathway, which not only proposes miR-765 as a novel potential therapeutic target for PTC, but also further revealed the associated mechanism.

Radioactive 125I seed implantation has definite curative effect in the treatment of local liver cancer. Currently, percutaneous implantation guided by CT, nuclear magnetism or ultrasound can be adopted, which is safe and minimally invasive. However, the images of some patients with metastatic liver cancer show isodensity or iso-echo imaging under CT or ultrasound, which is easy to miss diagnosis and treatment, seriously affecting the quality of life of patients. A case of terminal cholangiocarcinoma of the biliary tract with metastasis to the liver was admitted to China-Japan Union Hospital of Jilin University. Ultrasound image showed iso-echo, and radio 125I seed implantation was performed under the guidance of contrast-enhanced ultrasound. The operation was completed according to the preoperative treatment plan.

Objective:To investigate the effect of trifluoperazine (TFP) on the proliferation of mesangial cells through FoxO1 pathway and its protective effect on lupus nephritis (LN) mice.Methods:In vitro, MTT assay was used to detect the effect of different concentrations of trifluoperazine on the proliferation of mesangial cells. Flow cytometry was used to detect the effect of different concentrations of TFP on the mesangial cell cycle. Western blotting method (WB method) was used to detect the effect of different concentrations of TFP on the expression of FoxO1 and cyclinD1 proteins in mesangial cells, and the expression of TFP on mesangial cells, cyclinD1 and P21 after enhancing or inhibiting the expression of FoxO1. The effect of TFP on the proliferation of MRL/LPR mouse mesangial cells and the expression of FoxO1 was detected by HE staining, immunohistochemistry and WB method, and the effect of TFP on renal function in LN mice was detected by ELISA method in vivo. WB method was used to detect the effect of TFP on mesangial cell fibrosis, that is, the protein expression levels of FN and Col1. Repeated measures analysis of variance and One-way analysis of variance were used to compare measurement data between groups, and further pairwise comparisons were made using the Bonferroni method. Results:Trifluoperazine inhibits the proliferation of mesangial cells, and the interaction effects was concentration dependence and were statistically signifiant between groups, time ( Fgroup=162.58, Ftime=50.84, Finteraction=19.12, P<0.001). Flow cytometry results showed that after mesangial cells were treated with trifluoperazine at different concentrations, the percentage of cells in the G 0/G 1 phase gradually increased, while the cells in the S phase gradually decreased. This effect was dose-dependent, the difference was statistically significant ( P<0.05). Results of WB test proved that trifluoperazine inhibited the expression of cyclinD1 protein (2.17±0.34, 1.49±0.20, 1.11±0.27, 0.15±1.55, F=33.60 , P<0.001) and up-regulated the expression of FoxO1 (0.81±0.45, 2.31±0.81, 3.51±0.52, 5.13±10.07, F=35.63, P<0.001), and also in a dose-dependent patten. In vivo experimental results showed that trifluoperazine could inhibit the proliferation of mesangial cells and promote the expression of FoxO1 in mice with lupus nephritis, and the difference wsa statistically significant ( F=8.47, P=0.007). ELISA test results showed that trifluoperazine had a protective effect on renal function [serum creatinine: normal group(144±23)μmol/L, LN group (237±14)μmol/L, LN+TFP(211±36)μmol/L, Fvalue=20.47, P<0.001, urea nitrogen: normal group (22.84±0.56)μmol/L, LN group (19.99±0.92)μmol/L, LN+TFP (13.57±0.25)μmol/L, F=331.96, P<0.001] and it was proved by WB method that trifluoperazine could inhibit Fn and Col1 expression, the difference was statistically significant ( FFN=1 312.83, FCol1=171.16, P<0.001). Conclusion:Trifluoperazine blocks the mesangial cell cycle in G 0/G 1 phase by increasing the expression of FoxO1 and inhibits cell proliferation, which may have a therapeutic effect on lupus nephritis nephritis.

Enterotoxigenic Escherichia coli（ETEC）infection can induce watery diarrhea，leading to dehydration，electrolyte disturbance，and even death in severe cases. Recombinant B subunit/inactivated whole-cell cholera（rBS/WC）vaccine is effective in preventing ETEC infectious diarrhea. On the basis of the latest evidence on etiology and epidemiology of ETEC，as well as the effectiveness，safety，and health economics of rBS/WC vaccine，National Clinical Research Center for Child Health（The Children’s Hospital，Zhejiang University School of Medicine）and Zhejiang Provincial Center for Disease Control and Prevention invited experts to develop expert consensus on rBS/WC vaccine in prevention of ETEC infectious diarrhea. It aims to provide the clinicians and vaccination professionals with guidelines on using rBS/WC vaccine to reduce the incidence of ETEC infectious diarrhea.

Objective     To assess the clinical value of preoperative localization coupled with computed tomography (CT) three-dimensional reconstruction in pulmonary nodule-centered uniportal thoracoscopic combined subsegmental/segmental resection. Methods     The clinical data of 30 patients of combined subsegmental/segmental resection in our hospital from December 2019 to October 2021 were retrospectively collected. There were 19 males and 11 females with the mean age of 56.4 (32.0-71.0) years. The pulmonary nodules were located by CT-guided injection of glue before operation. The three-dimensional reconstruction image and operation planning were carried out by Mimics 21.0 software. Results    The operations were all successfully performed, and there was no conversion to open thoracotomy or lobectomy. The mean tumor diameter was 11.6±3.5 mm, the mean distance between the nodule and the visceral pleura was 13.6±5.6 mm, the mean width of the actual cutting edge was 25.0±6.5 mm, the mean operation time was 110.2±23.8 min, the mean number of lymph node dissection stations was 6.5±2.4, the mean amount of intraoperative bleeding was 50.8±20.3 mL, the mean retention time of thoracic catheter was 3.2±1.1 d, and the mean postoperative hospital stay was 4.5± 1.7 d. There was 1 patient of subcutaneous emphysema, 1 patient of atrial fibrillation and 1 patient of blood in sputum. Conclusion     Preoperative CT-guided injection of medical glue combined with CT three-dimensional reconstruction of pulmonary bronchus and blood vessels is safe and feasible in pulmonary nodule-centered uniportal thoracoscopic  combined subsegmental/segmental resection, which ensures the surgical margin and reserves lung tissues.

BACKGROUND: With the extensive application of segmental lung resection in the treatment of early-stage lung cancer, how to complete segmentectomy more accurately and minimally invasively has become a research hotspot. The aim of this study is to explore the application of three-dimensional computed tomography bronchography and angiography (3D-CTBA) combined with perfusion area recognition technique in single-hole thoracoscopic complex segmentectomy. METHODS: From January 2021 to January 2022, the clinical data of 112 consecutive patients undergoing single-port thoracoscopic complex segmentectomy in the Department of Thoracic Surgery, Xuanwu Hospital, Capital Medical University were retrospectively analyzed. The three-dimensional reconstruction combined with perfusion area identification technique was used to perform the operation and the clinical data were analyzed. RESULTS: The average operation time was (141.1±35.4) min; the initial time of intersegmental plane display was (12.5±1.7) s; the maintenance time of intersegmental plane was (114.3±10.9) s; the intersegmental plane was clearly displayed (100%); the amount of bleeding was [10 (10, 20)] mL; the total postoperative drainage volume was (380.5±139.7) mL; the postoperative extubation time was (3.9±1.2) d; and the postoperative hospitalization time was (5.2±1.6) d. Postoperative complications occurred in 8 cases. CONCLUSIONS The advantages of 3D-CTBA combined with perfusion area recognition technique are fast, accurate and safe in identifying intersegmental boundary in single-port thoracoscopic complex segmentectomy, which could provide guidances for accuratding resection of tumors, shortening operation time and reducing surgical complications.
Humans ; Lung Neoplasms/pathology* ; Bronchography ; Pneumonectomy/methods* ; Retrospective Studies ; Thoracic Surgery, Video-Assisted/methods* ; Tomography, X-Ray Computed ; Angiography/methods* ; Perfusion

Objective    To evaluate the learning curve of CT-guided medical glue localization for pulmonary nodule before video-assisted thoracic surgery (VATS). Methods    The clinical data of the patients with pulmonary nodules who underwent CT-guided medical glue localization before VATS in our hospital from July 2018 to March 2021 were retrospectively analyzed. The patients were divided into 3 groups: a group A (from July 2018 to August 2019), a group B (from September 2019 to June 2020) and a group C (from July 2020 to March 2021). The localization time, morbidity, complete resection rate and other indexes were compared among the three groups. Results    A total of 77 patients were enrolled, including 24 males and 53 females aged 57.4±10.1 years. There were 25 patients in the group A, 21 patients in the group B, and 31 patients in the group C. 77 pulmonary nodules were localized. There was no significant difference among the groups in the basic data (P>0.05). The localization time in the group C was 10.6±2.0 min, which was statistically shorter than that in the group A (15.4±4.4 min) and group B (12.9±4.3 min) (P<0.01). The incidence of complications in the group C was lower than that in the group A and group B (25.8% vs. 52.0% vs. 47.6%, P=0.04). The success rate of localization of the three groups was not statistically different (P=0.12). Conclusion    There is a learning curve in CT-guided medical glue localization for single pulmonary nodule before VATS. After the first 46 cases, the operation time can be shortened, and the incidence of complications can be decreased.