OBJECTIVE: To study the mode of cell death in the penile cavernosum tissue of male rats in the hypoandrogenic state. METHODS: We equally randomized 36 10-week-old SD male rats into six groups: 4-week sham-operation (4-wk SO), 8-week sham-operation (8-wk SO), 4-week castration (4-wk C), 8-week castration (8-wk C), 4-week castration + testosterone replacement (4-wk C+T), and 8-week castration + T replacement (8-wk C+T). The rats in the SO groups received incision of the scrotal skin with preservation of the testis and epididymis, those in the C groups underwent bilateral orchiectomy and epididymectomy, and those in the C+T groups were subcutaneously injected with T propionate at 3 mg/kg qd alt. We measured the ratio of maximum intracavernous pressure to mean arterial pressure (ICPmax/MAP), the concentration of serum T and the level of nitric oxide (NO), and determined the expressions of active caspase-1 and caspase-3 in the penile cavernosum of all the animals. RESULTS: The ICPmax/MAP ratio, serum T and NO levels and smooth muscle / collagen ratio were significantly lower in the C than in the SO and C+T groups (all P<0.05). The rats in the 4-wk C group, compared with those in the SO group, showed dramatic increases in the rates of endothelial cell pyroptosis (［15.31 ± 0.55］% vs ［0.78 ± 0.53］%, P<0.01), endothelial cell apoptosis (［16.32 ± 0.97］% vs ［0.88 ± 0.39］%, P<0.01), total cell pyroptosis (［9.67 ± 0.49］% vs ［1.53 ± 0.24］%, P<0.01) and total cell apoptosis (［11.27 ± 0.94］% vs ［1.68 ± 0.15］%, P<0.01) in the penile cavernous tissue, and so did those of the 8-wk C group in the rates of endothelial cell pyroptosis (［27.37 ± 0.65］% vs ［1.02 ± 0.65］%, P<0.01), endothelial cell apoptosis (［24.27 ± 0.54］% vs ［1.00 ± 0.63］%, P<0.01), total cell pyroptosis (［14.85 ± 0.55］% vs ［1.72 ± 0.52］%, P<0.01) and total cell apoptosis (［15.92 ± 0.53］% vs ［1.27 ± 0.31］%, P<0.01). The rats of the 8-wk C group exhibited an even more significant elevation than those of the 4-wk C group in the rates of endothelial cell pyroptosis (［27.37 ± 0.65］% vs ［15.31 ± 0.55］%, P<0.05) and endothelial cell apoptosis (［24.27 ± 0.54］% vs ［16.32 ± 0.97］%, P<0.05) . CONCLUSION Pyroptosis and apoptosis are the main modes of death of endothelial cells in the penile cavernosum of male rats after 4 weeks of castration and, with the progression of the disease, both increase with minimally expressed caspase-1 and caspase-3 in the smooth muscle cells. Endothelial cells and smooth muscle cells have different modes of death in different stages of hypoandrogenism.
Animals ; Male ; Penis/cytology* ; Rats ; Rats, Sprague-Dawley ; Apoptosis ; Caspase 3/metabolism* ; Caspase 1/metabolism* ; Testosterone/blood* ; Cell Death ; Orchiectomy

Circulating tumor DNA (ctDNA) is cell-free DNA released by tumors or circulating tumor cells, containing abundant tumor-specific information that can serve as biomarkers for cancer early screening, monitoring, prognosis, and prediction of treatment response. This is particularly attractive in the field of gastric cancer, where high-quality screening, monitoring, and prediction methods are currently lacking. Gastric cancer exhibits significant tumor heterogeneity, with large differences in genetic and epigenetic characteristics among different subgroups. Methylated ctDNA has high sensitivity and specificity, which can help clarify tumor genotyping and facilitate the formulation of precise diagnostic and therapeutic strategies. Furthermore, numerous studies have confirmed the unique advantages of methylated DNA in predicting treatment response, adjuvant therapy, and drug resistance assessment, which may be used in the future to enhance the efficacy of chemotherapy regimens and improve patient chemotherapeutic response, and even treat multidrug resistance. However, there are several challenges associated with methylated ctDNA, such as low sensitivity and specificity at single-target sites, limited association between some gastric cancer subtypes and ctDNA, off-target risks, and the lack of large-scale and high-quality clinical research evidence. This review mainly summarizes current research on the methylation status of ctDNA in gastric cancer and connects these findings to early screening, recurrence monitoring, and potential treatment opportunities for gastric cancer. With advances in technology and the deepening of interdisciplinary research, ctDNA detection will reveal more disease information and become an essential foundation for gastric cancer research and precision medicine treatment.

Objective To explore the relationship between soluble tumor necrosis factor-like weak inducer of apoptosis(sTWEAK),high-sensitivity C-reactive protein(hs-CRP)levels and the occurrence of major adverse cardiovascular events(MACE)in the short term after discharge in patients with acute myocardial infarction(AMI),and to construct a risk prediction model.Methods A total of 135 patients with AMI admitted to Nanyang Second General Hospital from October 2019 to October 2022 were selected as the research subjects.They were followed up for 6 months after discharge,and the occurrence of MACE was recorded.Patients with MACE were included in the MACE group,and the patients without MACE were included in the non-MACE group.The serum levels of sTWEAK and hs-CRP,as well as clinical data such as age,gender,body mass index(BMI),smoking history,drinking history,anemia,diabetes history,hypertension history,hyperlipidemia history,stroke history,old myocardial infarction,anterior wall infarction,Killip heart function classification,coronary artery lesion number,coronary artery Gensini score,acute physiology and chronic health evaluation Ⅱ(APACHE Ⅱ),white blood cell count,serum creatine kinase isoenzyme(CK-MB)level,serum cardiac troponin T(cTnT)level,and the time from onset to reperfusion were compared between the two groups.The risk factors for short-term MACE after discharge of AMI patients were analyzed by using a logistic regression model.Based on the logistic regression analysis results,a model for predicting the risk of short-term MACE after discharge of AMI patients was established,and its prediction efficiency and calibration capability were evaluated by using the receiver operating characteristic(ROC)curve and calibration curve.Results The incidence rate of MACE within 6 months after discharge in AMI patients was 27.73％(33/135).The proportion of patients over 60 years old,proportion of patients with smoking history,percentage of coronary artery lesions in two or more branches,APACHE Ⅱ score,and serum cTnT,sTWEAK and hs-CRP levels in the MACE group were significantly higher than those in the non-MACE group(P＜0.05).There were no statistically significant differences in gender distribution,BMI,drinking history,anemia,diabetes history,hypertension history,hyperlipidemia history,stroke history,old myocardial infarction,atrial fibrillation,anterior wall infarction,Killip heart function classification,time from onset to reperfusion,coronary artery Gensini score,white blood cell count,and serum CK-MB level between the MACE group and the non-MACE group(P＞0.05).Logistic regression model analysis showed that age over 60 years old,coronary artery lesions in two or more branches,high APACHE Ⅱ score,high serum cTnT level,high serum sTWEAK level,and high serum hs-CRP level were risk factors for the occurrence of short-term MACE after discharge of AMI patients(P＜0.05).A risk prediction model for MACE occurrence after discharge of AMI patients was constructed based on age,number of coronary artery branches with lesions,APACHE Ⅱ score,serum cTnT level,serum sTWEAK level,and serum hs-CRP level.The area under the curve of this model for predicting MACE was 0.860(95％confidence interval:0.784-0.916),with a sensitivity of 84.85％(95％confidence interval:68.100-94.900)and a specificity of 83.72％(95％confidence interval:74.200-90.800).The Hosmer-Lemeshow goodness-of-fit test showed there was no statistically significant difference between the predicted probability of short-term MACE occurrence after discharge of AMI patients and the actual probability(x2=5.541,P＞0.05).Conclusion High levels of serum sTWEAK and hs-CRP are risk factors for the occurrence of MACE after discharge in AMI patients.The risk prediction model constructed based on these two indicators has good performance for predicting MACE and can provide a reference for clinical guidance on the management and treatment of AMI patients after discharge.

Circulating tumor DNA (ctDNA) is cell-free DNA released by tumors or circulating tumor cells, containing abundant tumor-specific information that can serve as biomarkers for cancer early screening, monitoring, prognosis, and prediction of treatment response. This is particularly attractive in the field of gastric cancer, where high-quality screening, monitoring, and prediction methods are currently lacking. Gastric cancer exhibits significant tumor heterogeneity, with large differences in genetic and epigenetic characteristics among different subgroups. Methylated ctDNA has high sensitivity and specificity, which can help clarify tumor genotyping and facilitate the formulation of precise diagnostic and therapeutic strategies. Furthermore, numerous studies have confirmed the unique advantages of methylated DNA in predicting treatment response, adjuvant therapy, and drug resistance assessment, which may be used in the future to enhance the efficacy of chemotherapy regimens and improve patient chemotherapeutic response, and even treat multidrug resistance. However, there are several challenges associated with methylated ctDNA, such as low sensitivity and specificity at single-target sites, limited association between some gastric cancer subtypes and ctDNA, off-target risks, and the lack of large-scale and high-quality clinical research evidence. This review mainly summarizes current research on the methylation status of ctDNA in gastric cancer and connects these findings to early screening, recurrence monitoring, and potential treatment opportunities for gastric cancer. With advances in technology and the deepening of interdisciplinary research, ctDNA detection will reveal more disease information and become an essential foundation for gastric cancer research and precision medicine treatment.

OBJECTIVE: To study the preparation and properties of the hyaluronic acid (HA)/α-calcium sulfate hemihydrate (α-CSH)/β-tricalcium phosphate (β-TCP) material (hereinafter referred to as composite material). METHODS: Firstly, the α-CSH was prepared from calcium sulfate dihydrate by hydrothermal method, and the β-TCP was prepared by wet reaction of soluble calcium salt and phosphate. Secondly, the α-CSH and β-TCP were mixed in different proportions (10∶0, 9∶1, 8∶2, 7∶3, 5∶5, and 3∶7), and then mixed with HA solutions with concentrations of 0.1%, 0.25%, 0.5%, 1.0%, and 2.0%, respectively, at a liquid-solid ratio of 0.30 and 0.35 respectively to prepare HA/α-CSH/ β-TCP composite material. The α-CSH/β-TCP composite material prepared with α-CSH, β-TCP, and deionized water was used as the control. The composite material was analyzed by scanning electron microscope, X-ray diffraction analysis, initial/final setting time, degradation, compressive strength, dispersion, injectability, and cytotoxicity. RESULTS: The HA/α-CSH/β-TCP composite material was prepared successfully. The composite material has rough surface, densely packed irregular block particles and strip particles, and microporous structures, with the pore size mainly between 5 and 15 μm. When the content of β-TCP increased, the initial/final setting time of composite material increased, the degradation rate decreased, and the compressive strength showed a trend of first increasing and then weakening; there were significant differences between the composite materials with different α-CSH/β-TCP proportion ( P<0.05). Adding HA improved the injectable property of the composite material, and it showed an increasing trend with the increase of concentration ( P<0.05), but it has no obvious effect on the setting time of composite material ( P>0.05). The cytotoxicity level of HA/α-CSH/β-TCP composite material ranged from 0 to 1, without cytotoxicity. CONCLUSION The HA/α-CSH/β-TCP composite materials have good biocompatibility. Theoretically, it can meet the clinical needs of bone defect repairing, and may be a new artificial bone material with potential clinical application prospect.
Calcium Phosphates ; Bone and Bones ; Phosphates

Objective:To investigate the effects of pre-existing antibody on seroconversion rate after influenza vaccination.Methods:This study recruited 1 900 healthy volunteers to receive influenza split vaccines in Xinjiang Uygur Autonomous region and Yunnan Province from September 2009 to October 2018. Hemagglutinin agglutination inhibition assay was used to detect the titers of specific antibodies in blood samples collected before vaccination and 28 d after vaccination and the effects of pre-existing antibody on the seroconversion to different influenza vaccine components were analyzed.Results:Trend analysis showed that with the increasing titer of pre-existing antibody, the seroconversion rates to A/H1N1, A/H3N2, B/Victoria and B/Yamagata vaccine components were gradually decreased (χ 2=121.76, P<0.001; χ 2=67.58, P<0.001; χ 2=45.25, P<0.001; χ 2=54.55, P<0.001). After adjusting for factors such as region, gender and age, multivariate logistic regression showed that pre-existing antibody titer equal to or higher than 40 was an independent factor that affected the seroconversion to A/H1N1, A/H3N2 and B/Victoria vaccine components, and the adjusted OR (95%CI) values were 2.50(2.00-3.13)、1.64(1.35-2.00) and 2.50(1.79-3.45), respectively. Conclusions:The seroconversion rate to each vaccine component was negatively correlated with the pre-existing antibody titer. The factor that pre-existing antibody titer equal to or higher than 40 was detrimental to the seroconversion to A/H1N1, A/H3N2 and B/Victoria vaccine components, but had no significant influence on B/Yamagata seroconversion.

Objective:The antigenic and genetic characteristics of influenza A(H1N1)pdm09 virus isolated from the mainland of China during the 2018-2019 influenza surveillance year were analyzed.Methods:Two thousand nine hundred and fifty-eight influenza A(H1N1)pdm09 virus strains in the 2018-2019 influenza surveillance year were analyzed by hemagglutination inhibition test. The hemagglutinin(HA) gene of 279 influenza A(H1N1)pdm09 virus strains was sequenced and analyzed. The representative strains of the dominant clades were performed for antigenic characteristics using post-vaccination human antisera.Results:Two thousand eight hundred and sixty-one (97%, 2 861/2 958) viruses characterized were antigenically similar to A/Michigan/45/2015. All HA gene of the sequenced viruses belonged to 6B.1 clade, and 269(96%, 269/279) viruses belonged to 6B.1A subclade. Compared with the vaccine virus, it had the common amino acid substitutions of S74R, S164T and I295V in the HA protein. There were several small groups with common amino acid substitutions in the 6B.1A subclade, and 51% sequenced viruses had S183P amino acid substitution in this subclade. The result of antigenic analysis using post-vaccination human antiserums showed that most of the representative strains were well inhibited by the sera.Conclusions:The antigenicity of influenza A(H1N1)pdm09 viruses in the mainland of China in 2018-2019 influenza surveillance year matched well with the corresponding vaccine strain, but the HA gene had genetically diverse characteristic.

Objective Few studies are reported on the protective effect of valproic acid (VPA) against traumatic brain injury (TBI) by down-regulating the protein expressions of matrix metalloproteinase-9 (MMP-9) and aquaporin-4 (AQP-4) in the brain tissue. This study aimed to investigate the neuroprotective effects of different doses of VPA against TBI in experimental rats. Methods We randomly divided 100 adult male rats into five groups of equal number, sham operation, TBI model, and low- (30 mg), medium- (150 mg) and high-dose (300 mg) VPA treatment. At 1, 3, 7 and 14 days after modeling by controlled cortex impact, we obtained the modified Neurological Severity Scores (mNSS), measured the VPA concentration in the venous blood, and then killed the rats and harvested the brain tissue for determination of the water content using the dry-wet method and the expressions of MMP-9 and AQP-4 by Western blot and immunohistochemistry. Results At 1, 3, 7 and 14 days after modeling, the mNSSs in the high-dose VPA group were 4.6 ± 1.3, 3.8 ± 1.3, 3.0 ± 0.7 and 1.8 ± 0.8, respectively, significantly lower than 8.4 ± 0.9, 7.0 ± 0.7, 5.8 ± 1.0 and 4.5 ± 1.3 in the TBI group (P < 0.05), decreasing in a time-dependent manner, with statistically significant difference between any two dose groups (P < 0.05). At 1, 3 and 7 days, the water contents in the brain tissue were (76.2 ± 0.7)%, (76.9 ± 1.7)% and (73.9 ± 1.3)% in the high-dose VPA group, significantly lower than (79.6 ± 0.8)%, (82.6 ± 0.8)% and (78.6 ± 0.7)% in the TBI group (P < 0.05), also decreasing in a time-dependent manner, with statistically significant difference between any two dose groups (P < 0.05). At 1 and 3 days, the expressions of MMP-9 and AQP-4 in the brain tissue were markedly down-regulated in the VPA groups in a dose-dependent manner as compared with those in the TBI group (P < 0.05), with statistically significant difference between any two dose groups (P < 0.05), and meanwhile immunohistochemistry showed large numbers of cells with positive expressions of MMP-9 and AQP-4, which were reduced with the increased dose of VPA. Conclusion VPA has a neuroprotective effect against TBI in rats by inhibiting the expressions of MMP-9 and AQP-4 proteins in the brain tissue and alleviating brain edema. Within the range of the doses studied, higher-dose VPA produces a better effect.

Objective Mild hypothermia (MHT) can effectively protect the brain in traumatic brain injury (TBI). This study was to investigate the effects of MHT on the calmodulin (CAM) expression and brain edema in the rat model of TBI. Methods Ninety adult SD rats were randomly divided into a sham operation, a normal temperature and an MHT group of equal number. Immediately after TBI, the rats of the MHT group maintained at a rectal temperature of (32 ± 0.5) °C for 6 hours. Modified neurological severity scores (mNSS) were obtained from 6 rats in each group at 1, 3, 5 and 7 days after modeling, and the rest of the animals subjected to brain MRI at 6, 12, 24 and 48 hours and then killed for determination of the CAM gene transcription and protein expression in the brain tissue by real-time PCR, immunohistochemistry and Western blot. Results The mNSSs were significantly higher in the MHT and normal temperature groups than in the sham operation control (P < 0.05) at all time points, neurological severity markedly decreased in the MHT group compared with the normal temperature group (P < 0.05). At 6, 12, 24 and 48 hours, the expression of CAM mRNA was remarkably down-regulated in the MHT group (1.83 ± 0.19, 1.72 ± 0.12, 1.59 ± 0.06 and 1.60 ± 0.07) in comparison with the normal temperature group (2.76 ± 0.25, 2.49 ± 0.18, 2.04 ± 0.14 and 1.65 ± 0.09) (P < 0.05), even lower in the MHT than in the normal temperature group (P < 0.05), but higher in both of the two groups than in the sham operation group (P < 0.05). At 6, 12, 24 and 48 hours, the volume of brain edema was significantly reduced in the MHT group ([32.14 ± 4.52], [36.52 ± 4.10], [42.10 ± 4.38] and [46.25 ± 5.02] mm³) as compared with the normal temperature group ([48.56 ± 5.35], [53.13 ± 6.31], [59.23 ± 6.82] and [62.35 ± 7.25] mm³) (P < 0.05). Conclusion Mild hypothermia can improve the neurological function and reduce the CAM expression and brain edema in the brain tissue of rats with traumatic brain injury, which may be related to the neuroprotective effect of mild hypothermia.

Objective To analyze the frequency of interleukin (IL)-22+CD161+CD4+ T cells in the peripheral blood mononuclear cells (PBMCs) in rheumatoid arthritis (RA) patients compared with healthy control subjects and investigate the relationship of IL-22+CD4+CD161+ T lymphocyte frequency changes with RA disease activity.In addition to explore the pathogenesis of RA,and to look for new treatment targets for RA.Methods Twenty-one RA cases were included in the Department of Rheumatology of Tangshan Gongren Hospital from 2017 to 2018.Fourteen patients were female and 7 were male with the age ranged from 36 to 74 years old.The average age of this group of patients was (55±10) years,the average disease course was (60±50) months.All patients fulfilled the classification criteria of American College of Rheumatology [American College of Rheumatology (ACR)].Twenty-one subjects were enrolled as the control group,all of them came to Tangshan Gongren Hospital for regular health check-up.Fifteen subjects in the control group were female and 6 were male.Their age ranged between 40-78 years old with the average age of (55±9) years.IL-22+CD4+CD161+ T cells in PBMCs were detected by flow cytometry.The frequency variation of different CD4+CD161 + T was compared between case and control groups.The correlation was studied between the frequency and RA disease activity score (DAS28),tender joints number,swollen joints number,red blood cell sedimentation rate,high sensitive C reactive protein and white blood cell counts,red blood cell counts,platelet counts,IgG,IgA,IgM,complement C3 level,complement C4 level.T-test or Mann-Whitney U test were used for single-factor analysis,Pearson's test was used for correlation analysis.Results The percentage of RA group secreted CD4+ T cells (0.33± 0.20)％ of INF-γand IL-22,CD4+ T cells (0.51±0.29)％ of IL-22,and CD4+CD161+ T cells of IL-22 simultaneously.The number (0.55 ±0.28)％ was.significantly higher than that of the healtby control group [(0.22±0.14)％,(0.25±0.18)％,(0.36±0.24)％],and the differences were statistically significant [P=0.002,P=-0.0.45,P=0.026].Conclusion The percentage of IL-22+CD4+CD161+ T lymphocytes in the peripheral blood monocytes in RA patients is significantly higher than that in the healthy controls.The results of this study suggest that IL-22+CD4+CD161+ T lymphocytes in RA patients maybe related to RA disease activity and joint lesions.