ObjectiveTo observe the effects of Tongmai Kaiqiao pills on AMP-activated protein kinase （AMPK）/peroxisome proliferator-activated receptor gamma coactivator-1α （PGC-1α） signaling pathway and mitochondrial biogenesis in hippocampal tissue of rats with vascular cognitive impairment （VCI） and to investigate the potential mechanism of Tongmai Kaiqiao pills in improving cognitive impairment in rats with VCI. MethodsTwelve of 72 male SD rats were selected as the sham operation group， and the remaining rats were modelled using the modified 2VO method. The rats that were successfully modelled were divided into the model group， the high-dose group of Tongmai Kaiqiao pills （27.6 g·kg^-1）， the low-dose group of Tongmai Kaiqiao pills （13.8 g·kg^-1）， the combination group （27.6 g·kg^-1 Tongmai Kaiqiao pills + 25 mg·kg^-1 dorsomorphin）， and the donepezil hydrochloride group （0.45 g·kg^-1） according to the random number table method. After four weeks of continuous intraperitoneal injection of the corresponding drugs， the Morris water maze test was used to test the learning and memory ability of rats. Hematoxylin-eosin （HE） staining and Nissl staining were used to detect pathological changes in the hippocampus of the rats. The content of mitochondrial adenosine triphosphate （ATP） in the brain hippocampus was detected by colorimetry， and reactive oxygen species （ROS） level was detected in rat mitochondria by MitoSOX Red assay. Mitochondrial DNA copy number was detected by real-time fluorescent quantitative PCR （Real-time PCR）. Pathological changes in mitochondria were observed by transmission electron microscopy （TEM）， and AMPK， PGC-1α， phosphorylated AMP-activated protein kinase （p-AMPK）， nuclear respiratory factor 1 （Nrf1）， and mitochondrial transcription factor A （TFAM） protein expression in the hippocampus of the rats were detected by Western blot. ResultsCompared with those in the sham operation group， rats in the model group had a reduced number of platform crossings （P<0.01）， significantly prolonged evasion latency （P<0.01）， disorganized neuronal arrangement in the hippocampal region， widened gaps， and blurred nucleus membrane and nucleolus boundaries. The emergence of necrotic cells was visible. The color of the nissl bodies was light， and the number was reduced with severe loss. Mitochondria were atrophied， and cristae were lost. Severe damage was observed. The content of ROS was increased， and the level of ATP was decreased. mtDNA copy number decreased significantly （P<0.01）， and the protein expression of p-AMPK， PGC-1α， Nrf1， and TFAM decreased （P<0.05， P<0.01）. Compared with those in the model group， rats in the high-dose group of Tongmai Kaiqiao pills and donepezil hydrochloride group showed a shorter time to find the platform （P<0.01）， increased number of platform crossings （P<0.01）， restored mitochondrial morphology and structure of the hippocampal neurons， alleviated neuronal death， increased number of nissl bodies， weaken degree of injury， lower content of ROS， and significantly increased levels of ATP and number of copies of mtDNA （P<0.05， P<0.01）. In addition， there was increased protein expression of p-AMPK， PGC-1α， Nrf1， and TFAM （P<0.05， P<0.01）. Compared with the model group， the evasion latency was shortened in the low-dose group of Tongmai Kaiqiao pills （P<0.01）， and the number of platform crossings was increased， but the difference was not statistically significant. The mitochondria were swollen and deformed， and the cristae became shorter and partially disappeared. The degree of damage did not improve significantly， and the number of nissl bodies was increased but not statistically significant. The ROS content decreased （P<0.01）， but there was no significant difference in ATP level and mtDNA copy number. The protein expression of PGC-1α was increased （P<0.05）， but there was no significant difference in the protein expression of p-AMPK， Nrf1， and TFAM， and the results were not statistically significant. Compared with the donepezil hydrochloride group， there was no significant change in the results of each assay in the high-dose group of Tongmai Kaiqiao pills， and the difference was not statistically significant. Compared with the high-dose group of Tongmai Kaiqiao pills， rats in the combination group had a significantly lower number of platform crossings （P<0.01）， a significantly longer evasion latency （P<0.01）， a reduced number of neuronal cells， disorganized tissue structure， swollen and blurred cell outlines， a significant reduction in the number of nissl bodies. Moreover， there was an increase in the content of ROS， a decrease in the level of ATP and the number of mtDNA copies （P<0.01）， and a decrease in the expression of p-AMPK， PGC-1α， Nrf1， and TFAM （P<0.05）. ConclusionTongmai Kaiqiao pills is able to improve cognitive function in rats by activating the AMPK/PGC-1α signaling pathway， promoting mitochondrial biogenesis， and attenuating pathological damage to neurons in the hippocampal region， thereby demonstrating its therapeutic potential.

Objective To investigate the correlation between claudin-5 level and the risk of hemor-rhagic transformation(HT)in elderly stroke patients after thrombolysis.Methods A total of 192 elderly stroke patients hospitalized in our department from February 2021 to October 2022 were recruited,and divided into HT group(32 cases,including 14 of parenchymal hematoma type and 18 of hemorrhagic cerebral infarction type)and non-HT group(160 cases)according to the results of head CT or MRI 48 h after thrombolysis.The clinical data and serum claudin-5 level were com-pared between the two groups.Multivariate logistic regression analysis was used to identify the in-fluencing factors of HT risk after thrombolysis in elderly stroke patients.Restricted cubic splines were drawn to analyze the correlation between claudin-5 level and HT risk after thrombolysis.Re-sults The HT group had higher proportion of atrial fibrillation,longer onset to thrombolysis time,higher baseline NIHSS score,and fasting blood glucose and claudin-5 levels than the non-HT group(P＜0.05,P＜0.01).Multivariate logistic regression analysis showed that time from onset to thrombolysis(OR=2.589,95％CI:1.202-5.573,P=0.015),baseline NIHSS score(OR=1.415,95％CI:1.213-1.651,P=0.000),and fasting blood glucose(OR=1.552,95％CI:1.014-2.375,P=0.043)and claudin-5 levels(OR=1.174,95％CI:1.076-1.281,P=0.000)were risk factors for HT after thrombolysis in elderly stroke patients.Restricted cubic spline anal-ysis demonstrated that claudin-5 level had a nonlinear relationship with the risk of HT in the pa-tients after thrombolysis(x2=12.380,P=0.006).Conclusion Serum claudin-5 level is associated with HT after thrombolysis in elderly stroke patients,and shows a dose-response relationship with HT.

Objective To explore the diagnostic value of a wearable flexible patch ECG instrument in arrhythmia,the alarm situation during clinical application,patient satisfaction and safety.Methods A total of 1 443 subjects wore flexible patch ECG and conventional dynamic ECG(control)for 24 h to test the validity and consistency of arrhythmia diagnosis,and counted the alarm of remote ECG and the occurrence of related adverse events during the wearing of the instrument.Results There were 987 cases of arrhythmia detected by flexible patch ECG and 992 cases by conventional dynamic ECG.The total coincidence rate of arrhythmia diagnosis was 98.7%.The mean heart rate was measured by flexible patch ECG(75.4±11.4)times/min,conventional dynamic heart rate(71.5±12.1)times/min,the intra-group correlation coefficient(ICC)of 2 instruments was 0.892(95%CI:0.537-0.956),with good repeatability.The correct alarm rate of flexible patch ECG was 100%.The incidences of skin pruritus(0.28%vs.1.32%),skin allergy,redness and swelling(0.14%vs.0.69%)and electrode strip shedding(0 vs.0.28%)during wearing the flexible patch electrocardiogram were lower than those of the conventional holter electrocardiogram(P＜0.05).Conclusion The flexible patch ECG has few adverse reactions,high comfort,good safety and clinical applicability.

Vaccination is a main measure for protecting chickens against Marek's disease,while it is not able to suppress the infection,proliferation,transmission,and virulence enhancement on Marek's disease virus.Inhibiting the proliferation of Marek's disease virus in chicken is therefore an im-portant option for enhancing defense effectiveness.In this study,a compound,DUBs-IN-1,was found to inhibit the activity of MDV049,a protease encoded by Marek's disease virus,via screening a protease inhibitor library using MDV049 as target and ubiquitin probe.Molecular docking re-vealed that DUBs-IN-1 can interact with the residues which formed the catalytic pocket of MDV049,blocking the interaction between Ub substrate and the catalytic center of MDV049,then suppress the activity of MDV049 with competitive inhibition.Using the CPE model,it was found that DUBs-IN-1 at the concentration of 0.35 and 0.70 μmol/L significantly inhibited the CPE in-duced by Marek's disease virus in CEF cells.Quantitative analysis revealed that DUBs-IN-1 inhibi-ted the proliferation of Marek's disease virus in CEF cells(P＜0.01).Furthermore,it was found that the administration of 80 and 150 pg/(kg·d)of DUBs-IN-1 in chicken infected by Marek's disease virus significantly inhibited the proliferation of MDV in T cells(P＜0.01).In summary,this study demonstrated that the compound DUBs-IN-1 is able to inhibit the proliferation of Marek's disease virus in chicken cells,laying a theoretical and practical foundation for further de-velopment of the drugs against Marek's disease virus.

Background: Feline panleukopenia virus (FPV) is a widespread and highly infectious pathogen in cats with a high mortality rate. Although Yanji has a developed cat breeding industry, the variation of FPV locally is still unclear. Objectives: This study aimed to isolate and investigate the epidemiology of FPV in Yanji between 2021 and 2022. Methods: A strain of FPV was isolated from F81 cells. Cats suspected of FPV infection (n = 80) between 2021 and 2022 from Yanji were enrolled in this study. The capsid protein 2 (VP2) of FPV was amplified. It was cloned into the pMD-19T vector and transformed into a competent Escherichia coli strain. The positive colonies were analyzed via VP2 Sanger sequencing. A phylogenetic analysis based on a VP2 coding sequence was performed to identify the genetic relationships between the strains. Results: An FPV strain named YBYJ-1 was successfully isolated. The virus diameter was approximately 20–24 nm, 50% tissue culture infectious dose = 1 × 10 −4.94 /mL, which caused cytopathic effect in F81 cells. The epidemiological survey from 2021 to 2022 showed that 27 of the 80 samples were FPV-positive. Additionally, three strains positive for CPV-2c were unexpectedly found. Phylogenetic analysis showed that most of the 27 FPV strains belonged to the same group, and no mutations were found in the critical amino acids. Conclusions A local FPV strain named YBYJ-1 was successfully isolated. There was no critical mutation in FPV in Yanji, but some cases with CPV-2c infected cats were identified.

Long non-coding RNA (lncRNA) has become an important regulator of many cellular processes, including cell proliferation. Although studies have shown that a variety of lncRNAs play an important role in the occurrence and development of hematopoietic malignancies, a more comprehensive and unbiased method to study the function of lncRNAs in leukemia cell lines is lacking. Here, we used short hairpin RNA (shRNA) library combined with high-throughput sequencing to screen lncRNAs that may affect the proliferation of leukemia cell lines, and identified lncRNA C20orf204-203 among 74 candidate lncRNAs in this study. Further experiments showed that C20orf204-203 was localized in the cytoplasm in both K562 and THP-1 cell lines. C20orf204-203 knockdown decreased the proliferation of K562 and THP-1 cell lines accompanied with the increased proportion of early apoptotic cells. We observed the increased mRNA level of BAD gene while decreased protein level of TP53 and BCL2. The expression of Caspase 3 decreased and Caspase 3-cleaved protein increased in THP-1 cell line. However, their changes were inconsistent in the two cell lines. Our experimental results showed that knockdown of lncRNA C20orf204-203 in leukemia cell lines affected cell proliferation although the mechanism of action in different cell lines may differ. Importantly, our research demonstrated the feasibility of using shRNA library combined with high-throughput sequencing to study the role of lncRNA in leukemia cell lines on a large scale.
Caspase 3 ; Cell Line, Tumor ; Cell Proliferation/genetics* ; Humans ; Lentivirus/genetics* ; Leukemia/genetics* ; Proto-Oncogene Proteins c-bcl-2 ; RNA, Long Noncoding/metabolism* ; RNA, Messenger ; RNA, Small Interfering/genetics*

Objective:To investigate the risk factors affecting the occurrence of perioperative complications in patients with locally progressive gastric cancer undergoing radical gastric cancer treatment.Methods:The clinical data of 129 patients with locally progressive gastric cancer from January 2017 to December 2019 in Shaanxi Provincial People′s Hospital were retrospectively analyzed, including 98 males and 31 females, with an age ranged from 27 to 79 years and a mean age of (60.61±10.00) years. The postoperative complications of 129 patients with gastric cancer were firstly counted, and then the relationship between clinical data such as patients′ general condition, intraoperative status and pathological indexes and the occurrence of perioperative complications was analyzed by using univariate analysis, and significant factors were included in the logistic regression model for multifactor analysis to study the independent risk factors for the occurrence of perioperative complications.Results:Of the 129 patients, 25 cases (19.38%) had postoperative complications, including 10 cases (7.75%) with Clavien-Dindo classification combined with grade Ⅲ or higher complications. The results of univariate analysis suggested ACCI score >4 (30.76% vs 68.00%, χ2=11.86, P=0.001), body mass index ≥25 kg/m 2 (24.03% vs 60.00%, χ2=12.18, P=0.001), and preoperative hypoproteinemia (17.30% vs 36.00%, χ2=4.25, P=0.039), vascular cancer embolism (14.42% vs 40.00%, χ2=7.70, P=0.006), operative time ≥ 400 min (26.92% vs 52.00%, χ2=5.84, P=0.016), intraoperative bleeding ≥ 400 mL (13.46% vs 44.00%, χ2=12.03, P=0.001) were risk factors for the development of perioperative complications in patients with locally progressive gastric cancer. Multifactorial analysis showed that ACCI score >4, body mass index ≥25 kg/m 2, preoperative hypoproteinemia, vascular cancer embolism, and intraoperative bleeding ≥400 mL were independent risk factors for the occurrence of perioperative complications in patients with locally progressive gastric cancer ( P<0.05). Conclusions:The occurrence of perioperative complications in locally progressive gastric cancer hands was closely associated with ACCI score, body mass index, preoperative hypoproteinemia, vascular cancer embolism and intraoperative bleeding. ACCI score is expected to be a predictor of the occurrence of perioperative complications in patients with locally progressive gastric cancer.

Objective:To explore the dynamic changes of donor derived T cells at different time points in the aplastic anemia mouse model.Methods:The aplastic anemia mouse model was induced and then the proportion of infiltrated donor derived T cells in spleen and bone marrow, expression of activation molecular markers, cell cycle and functional subsets were measured by flow cytometry at different time points to evaluate the functional status of T cells in different periods.Results:①T cell immune-mediated aplastic anemia mouse model was successfully established by half lethal dose irradiation combined with major histocompatibility antigen (MHC) haploidentical lymph node cells infusion. ②The donor derived T cells began to infiltrate significantly in the spleen of aplastic anemia mouse from the 3rd day after transplantation and the ratio of CD4 +/CD8 + gradually inverted. After the 5th day, they gradually entered the bone marrow, predominated by CD8 + cells. ③The expression peak of CD69 in donor CD4 + cells was later than that in CD8 + cells. The trend of CD25 expression in CD4 + cells was the same as that in CD8 + cells, but the expression level in CD8 + cells was higher than CD4 + cells. ④The proportion of donor CD4 + cells in S/G 2/M phase reached the peak in spleen, about 12%, within 3 days after transplantation, while a higher level in CD8 + cells, which was about 20%. And the proportion of both CD4 + and CD8 + cells in S/G 2/M phase increased again after entering bone marrow, which was continued to be higher in CD8 + cells than that in CD4 + cells after 3 days of transplantation. ⑤Immune activated T cells in the spleen rapidly differentiated into effector memory T cells (T EM) after a short central memory T cell (T CM) stage. After entering the bone marrow, some T EM differentiated into effector cells to further function. Conclusion:In the aplastic anemia mouse model, donor derived T cells activated rapidly after entering the allogenic recipient, reached its proliferation booming period and differentiated into T EM cells within 5 days. After 5 days, they began to enter the bone marrow to continue proliferate and damage hematopoiesis.

Objective:To investigate the risk factors for anterior circulation and posterior circulation symptomatic intracranial atherosclerotic stenosis (sICAS).Methods:The clinical data of patients admitted to Hebei General Hospital for ischemic stroke or transient ischemic attack (TIA) and diagnosed with sICAS by digital subtraction angiography from May 2019 to May 2020 were retrospectively included. The patients were divided into anterior circulation group and posterior circulation group according to the stenosis sites, and the distribution of sICAS and its risk factors were analyzed.Results:A total of 134 patients with sICAS were enrolled, including 82 males (61.2%) and 52 females (38.8%). Their age was 60.28±11.46 years; 115 (85.8%) had ischemic stroke and 19 (14.2%) had TIA. There were 92 patients (68.7%) in the anterior circulation group and 42 (31.3%) in the posterior circulation group. Body mass index (BMI), systolic and diastolic blood pressure levels, as well as the proportion of patients with hypertension, diabetes, smoking and drinking in the posterior circulation group were significantly higher than those in the anterior circulation group (all P<0.05). Multivariate logistic regression analysis showed that higher BMI (odds ratio [ OR] 1.191, 95% confidence interval [ CI] 1.029-1.379; P=0.019), hypertension ( OR 4.073, 95% CI 1.135-14.616; P=0.031) and diabetes ( OR 2.783, 95% CI 1.149-6.738; P=0.023) were independently correlated with the posterior circulation sICAS. Conclusions:Compared with anterior circulation, high BMI, hypertension and diabetes are the independent risk factors for posterior circulation sICAS.

Homocysteine participates in the methionine cycle in the body and is excreted by the kidneys. Its abnormal elevation increases the risk of cerebrovascular disease. Studies in recent years have shown that hyperhomocysteinemia is closely related to cerebral small vessel diseases and is an independent risk factor for ischemic stroke and cognitive impairment. Because hyperhomocysteinemia is largely reversible, the significance of drug intervention in the prevention and treatment of cerebrovascular disease has gradually been confirmed. This article reviews the relationship between homocysteine and cerebral small vessel diseases in order to provide a reference for the prevention of stroke and dementia.