Secondary white matter injury after ischemic stroke refers to the secondary changes in the white matter structure outside the infarct site that are not directly damaged after stroke, often involving key connecting areas such as the corpus callosum and contralateral internal capsule. This injury is closely associated with post-stroke cognitive impairment, motor dysfunction, and language disorders, which affects the long-term outcome. Its pathophysiological mechanisms mainly include immune and inflammatory imbalance, excitotoxicity and mitochondrial dysfunction, blood-brain barrier damage, axonal injury, and remyelination disorder. Exploring these mechanisms in depth can help promote early diagnosis and intervention of secondary white matter damage after ischemic stroke, provide theoretical basis for the development of targeted treatment strategies, and promote functional recovery in patients with stroke.

Objective:To investigate the correlation between serum proprotein convertase subtilisin/Kexin type 9 (PCSK9) level and white matter hyperintensities (WMHs) in healthy population.Methods:Consecutive healthy individuals underwent routine physical examinations at the Department of Neurology, Jinling Hospital Affiliated to Medical School of Nanjing University (April 2023 to December 2023) and Hexi Branch of Nanjing First Hospital (March 2024 to April 2024) were included prospectively. Enzyme-linked immunosorbent assay was used to detect serum PCSK9 level. The Fazekas scale was used to assess the severity of WMHs (total score 0-6) and they were divided into no or mild WMHs group (0-2) and moderate to severe WMHs group (3-6). Multivariate logistic regression analysis was used to determine the independent correlation between the serum PCSK9 level and the severity of WMHs. Results:A total of 177 subjects were enrolled, including 110 males (62.1%), aged 66.7±10.1 years. The median serum PCSK9 level was 203.9 ng/L. According to the Fazekas score, there were 102 patients (51.6%) in the no or mild WMHs group, and 75 (42.4%) in the moderate to severe WMHs group. One way analysis of variance showed that serum PCSK9 level significantly increased with the increase of WMHs total score ( P=0.001). The serum PCSK9 level in the moderate to severe WMHs group was significantly higher than that in the no or mild WMHs group (437.2±260.4 ng/L vs. 217.9±141.7 ng/L; P=0.001). Multivariate logistic regression analysis showed that after adjusting for age, gender, and other confounding factors, there was a significant independent correlation between higher serum PCSK9 level and moderate to severe WMHs (odds ratio 3.201, 95% confidence interval 2.107-5.082; P=0.001). Conclusion:Higher serum PCSK9 level is an independent risk factor for moderate to severe WMHs in healthy individuals.

Objective:To evaluate the efficacy and safety of mitoxantrone hydrochloride liposome injection in the treatment of peripheral T-cell lymphoma (PTCL) in a real-world setting.Methods:This was a real-world ambispective cohort study (MOMENT study) (Chinese clinical trial registry number: ChiCTR2200062067). Clinical data were collected from 198 patients who received mitoxantrone hydrochloride liposome injection as monotherapy or combination therapy at 37 hospitals from January 2022 to January 2023, including 166 patients in the retrospective cohort and 32 patients in the prospective cohort; 10 patients in the treatment-na?ve group and 188 patients in the relapsed/refractory group. Clinical characteristics, efficacy and adverse events were summarized, and the overall survival (OS) and progression-free survival (PFS) were analyzed.Results:All 198 patients were treated with mitoxantrone hydrochloride liposome injection for a median of 3 cycles (range 1-7 cycles); 28 cases were treated with mitoxantrone hydrochloride liposome injection as monotherapy, and 170 cases were treated with the combination regimen. Among 188 relapsed/refractory patients, 45 cases (23.9%) were in complete remission (CR), 82 cases (43.6%) were in partial remission (PR), and 28 cases (14.9%) were in disease stabilization (SD), and 33 cases (17.6%) were in disease progression (PD), with an objective remission rate (ORR) of 67.6% (127/188). Among 10 treatment-na?ve patients, 4 cases (40.0%) were in CR, 5 cases (50.0%) were in PR, and 1 case (10.0%) was in PD, with an ORR of 90.0% (9/10). The median follow-up time was 2.9 months (95% CI 2.4-3.7 months), and the median PFS and OS of patients in relapsed/refractory and treatment-na?ve groups were not reached. In relapsed/refractory patients, the difference in ORR between patients with different number of treatment lines of mitoxantrone hydrochloride liposome injection [ORR of the second-line, the third-line and ≥the forth-line treatment was 74.4% (67/90), 73.9% (34/46) and 50.0% (26/52)] was statistically significant ( P = 0.008). Of the 198 PTCL patients, 182 cases (91.9%) experienced at least 1 time of treatment-related adverse events, and the incidence rate of ≥grade 3 adverse events was 66.7% (132/198), which was mainly characterized by hematologic adverse events. The ≥ grade 3 hematologic adverse events mainly included decreased lymphocyte count, decreased neutrophil count, decreased white blood cell count, and anemia; non-hematologic adverse events were mostly grade 1-2, mainly including pigmentation disorders and upper respiratory tract infection. Conclusions:The use of mitoxantrone hydrochloride liposome injection-containing regimen in the treatment of PTCL has definite efficacy and is well tolerated, and it is a new therapeutic option for PTCL patients.

Objective:To investigate the correlation between the lipocalin-2 (LCN-2) level and white matter hyperintensities (WMHs) in patients with ischemic stroke.Methods:Consecutive patients with ischemic stroke admitted to the Department of Neurology, Jinling Hospital, Medical School of Nanjing University from September 2021 to November 2021 and whose duration from onset to hospitalization <14 d were prospectively enrolled. Enzyme-linked immunosorbent assay was used to detect the serum LCN-2. Fazekas scale was used to assess the severity of periventricular and subcortical WMHs. A total WMHs score ≥3 was defined as severe WMHs. Multivariate logistic regression analysis was used to determine the correlation between serum LCN-2 level and WMHs. Results:A total of 179 patients were enrolled, including 122 males (68.2%), aged 64.7±11.6 years. The median serum LCN-2 level was 387.1 g/L, and 86 patients (48.0%) had severe WMHs. Serum LCN-2 in the severe WMH group was significantly higher than that in the non-severe WMH group (505.3±342.4 g/L vs. 367.8±224.5 g/L; t=3.110, P=0.002). Multivariable logistic regression analysis showed that after adjusting for the relevant confounding factors, there was a significant correlation between higher serum LCN-2 and severe WMHs (odds ratio 2.32, 95% confidence interval 1.17-4.63; P=0.017) and higher total WMHs score (odds ratio 1.62, 95% confidence interval 1.12-2.35; P=0.011). Conclusion:Higher serum LCN-2 level is associated with severe WMHs in patients with ischemic stroke.

Objective:To ascertain the basic situation of radiotherapy in Fujian in 2016.Methods:Based on the unified questionnaire, the types and quantity of radiotherapy and its corollary equipment, the number of radiotherapy staff and the work for quality control were surveyed in the radiotherapy units in the whole province, and the data on patients undergoing radiotherapy and other types of patients were collected from 26 hospitals. The total number of radiotherapy patients in the whole province was estimated by the correlation analysis and multiple linear regression analysis.Results:Radiotherapy was performed in a total of 32 hospitals in Fujian province in 2016. Among them, there were 62 sets of radiotherapy equipment, 33 sets of the simulators, 57 sets of treatment planning systems and 762 workers. The total number of 15 156 radiotherapy patients in 26 hospital were available for the survey. Multiple linear regression models showed that the frequency of application of medical electron accelerator was positively correlated with number of outpatients, emergency patients and inpatients, number of radiotherapy staff, number of electron accelerators ( r=0.311, 0.893, 0.956, P<0.05). Meanwhile, the frequency of aterloading brachytherapy was positively correlated with number of outpatients, emergency patients and inpatients, number of radiotherapy staff, number of aterloading brachytherapy units ( r=0.307, 0.966, 0.988, P<0.05). The frequency of radiotherapy was 0.54 patients per 1 000 population in Fujian in 2016. The delivered dose calibration of accelerators was performed in all hospitals involved in line with the relevant regulations, but the number of quality control equipment for radiotherapy was insufficient, such as the QA beam checker or well-type ionization chamber. Conclusions:In recent years, the rapid development of radiotherapy has been seen in Fujian province. The radiotherapy management should focus on standardenized and improved quality control and regulation in future.

OBJECTIVETo investigate the influence of siRNA-mediated down-regulation of CD147 on growth, proliferation and movement of human breast cancer cell line MDA-MB-231.

METHODSThe protein expression of CD147, MMP-2 and TIMP-2 of the MDA-MB-231 cells were analyzed by ABC. Lentiviral expression vector of CD147 gene was constructed and transfected into MDA-MB-231 cells. RT-PCR and Western blot were used to detect the mRNA and protein level changes of CD147 genes to identify the optimal time point, followed by detection of changes of mRNA and protein expression of MMP-2 and TIMP-2 genes. CCK-8 reagent method and cell scratch test were used to detect the proliferation and migration change of MDA-MB-231 cells. The nude mouse model of breast cancer by hypodermic injection with MDA-MB-231 cells was established to document the effect of CD147 siRNA on the tumor transplants.

RESULTSAfter transfection of lentiviral expression vector of CD147 gene, protein of CD147, MMP-2 and TIMP-2 were weakly or negative expressed, significantly weaker than those of control group (P < 0.01). After 72 hours of transfection, average down-regulation rate of CD147 and MMP-2 were 96.03% ± 0.84% and 96.03% ± 0.84%, respectively. Both CD147 mRNA and MMP-2 mRNA expression were down-regulated (P < 0.05), while TIMP-2 mRNA expression showed no significant deference (P > 0.05). No less than 2 days after transfection, cell growth of MDA-MB-231 cell line was found significantly inhibited (P < 0.05). After 24 hours of transection, average migration distance of MDA-MB-231 cell line and control group were (0.64 ± 0.12) mm and (4.69 ± 0.85) mm, respectively, which indicated a lower migrate speed. Down regulation of CD147 led to reduction of volume and mass of nude mouses. The growth of the carcinoma transplant was inhibited upon siRNA-mediated down-regulation of CD147 (P < 0.05), with an average tumor mass of (1.85 ± 0.98) g and both reduction of tumor size and tumor mass.

CONCLUSIONSCD147 may alter the MMP-2/TIMP-2 balance in MDA-MB-231 cells. CD147 gene silencing inhibits the proliferation and migration of MDA-MB-231 cells and the growth of carcinoma transplants in nude mice.

Animals ; Basigin ; metabolism ; Blotting, Western ; Breast Neoplasms ; genetics ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Down-Regulation ; Gene Expression Regulation, Neoplastic ; Gene Silencing ; Humans ; Matrix Metalloproteinase 2 ; metabolism ; Mice ; Mice, Nude ; Neoplasm Transplantation ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Tissue Inhibitor of Metalloproteinase-2 ; metabolism ; Transfection

Objective To investigate the influence of siRNA-mediated down-regulation of CD147 on growth, proliferation and movement of human breast cancer cell line MDA-MB-231.Methods The protein expression of CD147, MMP-2 and TIMP-2 of the MDA-MB-231 cells were analyzed by ABC.Lentiviral expression vector of CD 147 gene was constructed and transfected into MDA-MB-231 cells.RT-PCR and Western blot were used to detect the mRNA and protein level changes of CD 147 genes to identify the optimal time point , followed by detection of changes of mRNA and protein expression of MMP-2 and TIMP-2 genes. CCK-8 reagent method and cell scratch test were used to detect the proliferation and migration change of MDA-MB-231 cells.The nude mouse model of breast cancer by hypodermic injection with MDA-MB-231 cells was established to document the effect of CD 147 siRNA on the tumor transplants.Results After transfection of lentiviral expression vector of CD 147 gene, protein of CD147, MMP-2 and TIMP-2 were weakly or negative expressed , significantly weaker than those of control group ( P<0.01 ) .After 72 hours of transfection, average down-regulation rate of CD147 and MMP-2 were 96.03%±0.84% and 96.03%± 0.84%, respectively.Both CD147 mRNA and MMP-2 mRNA expression were down-regulated(P<0.05), while TIMP-2 mRNA expression showed no significant deference ( P >0.05 ) .No less than 2 days after transfection, cell growth of MDA-MB-231 cell line was found significantly inhibited ( P <0.05 ) .After 24 hours of tranfection, average migration distance of MDA-MB-231 cell line and control group were (0.64 ± 0.12)mm and (4.69 ±0.85)mm, respectively, which indicated a lower migrate speed.Down regulation of CD147 led to reduction of volume and mass of nude mouses .The growth of the carcinoma transplant was inhibited upon siRNA-mediated down-regulation of CD147 ( P <0.05 ) , with an average tumor mass of (1.85 ±0.98) g and both reduction of tumor size and tumor mass .Conclusions CD147 may alter the MMP-2/TIMP-2 balance in MDA-MB-231 cells. CD147 gene silencing inhibits the proliferation and migration of MDA-MB-231 cells and the growth of carcinoma transplants in nude mice .

OBJECTIVETo study the influence of CD147 gene silencing on the expression of ANXA2, MMP-2 and TIMP-2 of thyroid medullary carcinoma TT cells and related biological characteristics.

METHODSProtein expression of CD147, ANXA2, MMP-2 and TIMP-2 was detected by immunocytochemistry.RNAi technology was used to identify the specific siRNA sequences and the optimal time point of effective inhibition of CD147 gene. The expression of ANXA2, MMP-2 and TIMP-2 at mRNA and protein levels was detected with RT-PCR and Western blot, respectively. MTT method was used to detect the proliferation of the TT cells, flow cytometry (FCM) to detect the cell cycle and apoptosis changes of TT cells and transwell chamber assays to document the influence of CD147 gene silencing on migration and invasion of the TT cells.

RESULTSThe protein expression of CD147, ANXA2, MMP-2 and TIMP-2 proteins was variable in the TT cells. Two siRNA sequences were identified to effectively silence CD147 gene in the TT cells, in which relative expression of MMP-2 was reduced at both mRNA and protein levels; although the expression of ANXA2 mRNA and protein did not change significantly. TIMP-2 protein expression markedly decreased in an absence of its mRNA expression. The proliferation of the TT cells was inhibited upon the CD147 gene silencing along with a significant increase of G(0)/G(1) phase cells and a decrease of G(2)/M phase cells.However, the proportion of the apoptotic cells in all experimental groups did not change. The number of the penetrating cells through the membrane filters did not show significant changes in all experimental groups in the Transwell chamber assays.

CONCLUSIONSThrough RNAi technology, two CD147 siRNA sequences are identified and shown to effectively inhibit CD147 gene expression of the TT cells. CD147 gene silencing leads to growth inhibition of the TT cells and alteration of the cell cycle. However, silencing CD147 does not significantly affect the apoptosis, migration and invasion of the TT cells.

Annexin A2 ; genetics ; metabolism ; Basigin ; genetics ; metabolism ; Carcinoma, Medullary ; metabolism ; pathology ; Cell Cycle ; Cell Proliferation ; Gene Expression Regulation, Neoplastic ; Gene Silencing ; Humans ; Matrix Metalloproteinase 2 ; genetics ; metabolism ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Thyroid Neoplasms ; metabolism ; pathology ; Tissue Inhibitor of Metalloproteinase-2 ; genetics ; metabolism ; Transfection ; Tumor Cells, Cultured

Objective:This study aims to examine the clinicopathological features, diagnosis, and treatment of pulmonary margin-al zone B-cell lymphoma of mucosa-associated lymphoid tissue (PMZL-MALT). Methods:The clinicopathological features and immu-nohistochemical staining of CD20, CD79a, CD5, CD10, CD23, CyclinD1, and Ki-67 in seven patients with PMZL-MALT were ana-lyzed. Results:These patients, with a median age of 58 years, included three males and four females. Most of the patients suffered from cough, anhelation, and irregular fever. No specific imaging manifestation was observed. Tumor cells were positive for CD19 and CD20 but negative for CD5, CD10, and CyclinD1. The positive rate of Ki-67 was low. Conclusion:PMZL-MALT cases are easily misdiag-nosed because of the absence of specific clinical characteristics and X-ray features. Final diagnosis depends on pathological examina-tions.

Objective To investigate the distribution of TCR Vβ genealogy and clonal expansion in peripheral blood after infusing mesenchymal stem cells (MSC) in patients with chronic GVHD. Methods The complementarity determining region 3 (CDR3) of 24 TCR Vβ subfamily genes in peripheral blood mononuclear cell from 1 case with cGVHD after allogeneic hematopoietic stem cell transplantation (Allo-HSCT),who were treated with infusing MSC,were amplified using RT-PCR. The blood samples were taken at the first and the fifth day after 1st infusion; and the first day,the 10 th day and the 20 th day after the second infusion of MSC,as well as the MSC infused as control . The products were labelled by fluorescein and then analyzed the CDR3 size with gene scan technique to determine the clonality of T cells. Results There were no expression of TCR Vβ subfamily with the MSC infused and after the 1st day of the first infusion of MSC. Then 3,10,14,10 Vβ subfamilies clones are appeared at the other time points,of which were polyclone and oligoclone predominately. In the same time,the manifestations of cCVHD have been abated. Conclusion MSC played a certain role in reviving the immune function of the patients after Allo-HSCT and mitigating the disease of chronic GVHD. Lineage analysis of TCR Vβ subfamily showed some predominant expression.