ObjectiveTo investigate the effect of Toxoplasma gondii infection on copper metabolism in the kidneys of mice. MethodsA total of 80 7-8-week-old C57BL/6 female mice were randomly divided into four groups of 20 mice in each group after one week of adaptation， including Control group， Cu group， TgCtwh6 group and Cu+TgCtwh6 group. Mice that were not infected and fed with normal diet and water were used as the Control group； Mice fed with 1 g/kg of copper chloride processing diet and 0.1% copper chloride water for 60 consecutive days were used as Cu group； Mice infected with 25-30 TgCtwh6 cysts （one of the predominant genotype Chinese 1 in China） fed with normal diet and water were used as the TgCtwh6 group； mice infected with 25-30 TgCtwh6 cysts and fed with a processed diet containing 1 g/kg of copper chloride and water with 0.1% copper chloride for 60 consecutive days were used as the Cu+TgCtwh6 group. ICP-MS was used to determine the changes in copper content in kidney tissues. Hematoxylin-eosin （HE） staining was used to observe the pathological changes of mouse kidney tissue. The number of apoptotic cells was observed by PI staining. Western blot was used to detect the protein expression levels of glutathione peroxidase 4 （GPX4） and superoxide dismutase （SOD1， SOD2）. RT-qPCR was used to detect the mRNA expression of cuproptosis-related genes. ResultsPathological manifestations such as inflammatory cell infiltration in the Cu group and TgCtwh6 group were seen under the microscope， and the inflammatory infiltrating cells of the renal interstitial were reduced in the Cu+TgCtwh6 group， and the pathological manifestations

Objective:To analyze the genetic evolution characteristics of hemagglutinin (HA) and neuraminidase (NA) of influenza B virus in Tongling during 2019-2022 surveillance years.Methods:Twenty-two strains of Victoria influenza B virus isolated from our laboratory during 2019-2022 were selected for whole genome sequencing. The sequence comparison and phylogenetic analysis were conducted by using bioinformatic analysis software.Results:During 2019-2022, seasonal influenza in Tongling City was predominantly caused by influenza B Victoria lineage viruses, which fell within the V1A.3 branch. Among these, 14 strains isolated in the 2021-2022 season were further classified into the V1A.3a.2 sub-branch. Compared with vaccine strains, multiple amino acid mutation sites were detected in both HA and NA proteins of the 22 influenza B Victoria lineage viruses. Notably, all four major antigenic sites (120-loop, 150-loop, 160-helix, and 190-helix regions) in the HA protein exhibited variations. Although no mutations were detected at resistance sites on the NA protein, a change occurred in the glycosylation site at position 197 NETQ in the HA protein.Conclusions:The main amino acid sites of the HA protein of the influenza B Victoria lineage viruses in Tongling City from 2019 to 2022 have undergone significant variation, which may lead to antigenic drift. Therefore, it is essential to strengthen the monitoring of influenza virus mutations.

Direct conversion of cardiac fibroblasts (CFs) to cardiomyocytes (CMs) in vivo to regenerate heart tissue is an attractive approach. After myocardial infarction (MI), heart repair proceeds with an inflammation stage initiated by monocytes infiltration of the infarct zone establishing an immune microenvironment. However, whether and how the MI microenvironment influences the reprogramming of CFs remains unclear. Here, we found that in comparison with cardiac fibroblasts (CFs) cultured in vitro, CFs that transplanted into infarct region of MI mouse models resisted to cardiac reprogramming. RNA-seq analysis revealed upregulation of interferon (IFN) response genes in transplanted CFs, and subsequent inhibition of the IFN receptors increased reprogramming efficiency in vivo. Macrophage-secreted IFN-β was identified as the dominant upstream signaling factor after MI. CFs treated with macrophage-conditioned medium containing IFN-β displayed reduced reprogramming efficiency, while macrophage depletion or blocking the IFN signaling pathway after MI increased reprogramming efficiency in vivo. Co-IP, BiFC and Cut-tag assays showed that phosphorylated STAT1 downstream of IFN signaling in CFs could interact with the reprogramming factor GATA4 and inhibit the GATA4 chromatin occupancy in cardiac genes. Furthermore, upregulation of IFN-IFNAR-p-STAT1 signaling could stimulate CFs secretion of CCL2/7/12 chemokines, subsequently recruiting IFN-β-secreting macrophages. Together, these immune cells further activate STAT1 phosphorylation, enhancing CCL2/7/12 secretion and immune cell recruitment, ultimately forming a self-reinforcing positive feedback loop between CFs and macrophages via IFN-IFNAR-p-STAT1 that inhibits cardiac reprogramming in vivo. Cumulatively, our findings uncover an intercellular self-stimulating inflammatory circuit as a microenvironmental molecular barrier of in situ cardiac reprogramming that needs to be overcome for regenerative medicine applications.
Animals ; Mice ; Macrophages/immunology* ; Fibroblasts/cytology* ; Cellular Reprogramming ; STAT1 Transcription Factor/genetics* ; Signal Transduction ; Interferon-beta/genetics* ; Myocardial Infarction/pathology* ; Myocytes, Cardiac/cytology* ; Mice, Inbred C57BL ; GATA4 Transcription Factor/genetics* ; Male ; Cells, Cultured

Objective　To understand the impact of early and timely treatment and initial antiviral treatment regimen on mortality and attrition of antiretroviral therapy. Methods A retrospective cohort study was conducted using download data on antiretroviral therapy for HIV-infected patients in Liuzhou City, Guangxi Province, from the database of the Basic Information System for AIDS Control and Prevention (BISAC) from 2010 to 2020. The Cox proportional risk regression model was used to analyze the influencing factors of mortality and attrition. Results A total of 15 713 infected patients were included, including 53.4% aged 18-<50 years, 69.4% male, 61.0% farmer, 75.1% CD4 count <350 cells /μL before initial antiviral treatment, the overall mortality rate was 4.30/100 person-years, and the overall attrition was 2.42/100 person-years. The results of Cox regression analysis showed that the influencing factors of mortality were pretreatment CD4 counts of 350-<500 cells/μL(AHR=0.72, 95%CI: 0.63-0.81) and ≥500 cells/μL (AHR= 0.64, 95%CI: 0.55-0.76); duration from diagnosis to initial antiviral treatment 91-180 days (AHR=1.25, 95%CI: 1.08-1.45), 181-365 days (AHR=1.26, 95%CI: 1.08-1.47), and ≥365 days (AHR=1.26, 95%CI: 1.11-1.44); initial antiviral treatment regimens of D4T+3TC+EFV/NVP (AHR=1.47, 95%CI: 1.32-1.63) and AZT/D4T/TDF+3TC+LPV/r (AHR=1.73, 95%CI: 1.50-1.99). Factors affecting attrition were pretreatment CD4 counts of 350-499 cells/μL (AHR=1.32, 95%CI: 1.16-1.50) and ≥500 cells/μL (AHR=1.28, 95%CI: 1.10-1.50); interval from HIV positivity confirmation to initial dosing ≥365 days (AHR=1.21, 95%CI: 1.04-1.40), initial antiviral treatment regimens of TDF+3TC+NVP (AHR=1.32, 95%CI: 1.13-1.55), AZT+3TC+EFV/NVP (AHR=1.43, 95%CI: 1.26-1.62) and AZT/D4T/TDF+3TC+LPV/r (AHR=1.33, 95CI%: 1.06-1.67). Conclusions　Early and timely treatment and the initial antiviral treatment regimen of TDF+3TC+EFV have good efficacy, but attention should be paid to the high risk of attrition of HIV-infected people with high CD4 count before treatment.

Objective : To investigate the changes of iron and other metal elements and inflammatory factor γ interferon (IFN⁃γ) in the liver after taking deferiprone (DFP) in mice with chronic infection of Toxoplasma gondii , and to investigate the efficacy of DFP in the treatment of liver injury caused by chronic infection of Toxoplasma gondii. Methods : 6 ⁃week⁃old female C57BL/6 mice were randomly divided into four groups : control group , control + DFP group , TgCtWh6 group , TgCtWh6 + DFP group , with 6 mice in each group , and sterile gavage PBS in control group. Daily gavage was 50 μg/kg DFP in TgCtWh6 + DFP group , 20 toxoplasma cysts in TgCtWh6 group , 20 toxoplasmosis cysts in TgCtWh6 + DFP group , 20 toxoplasmosis cysts with daily gavage of 50 μg/kg DFP , and mice were sacrificed and liver tissues were taken after 45 days. Inductively coupled plasma mass spectrometry (ICP⁃MS) was used to detect the content of iron and other metal elements in mouse liver tissues. Real⁃time PCR (qRT⁃PCR) and Western blot were used to detect the mRNA levels and protein levels of IFN⁃γ and ferritin heavy chain (Fth) in mouse liver tissues , respectively. Results : Compared with the TgCtWh6 group , the Fth content in the liver (P < 0. 01) was significantly reduced in the liver of the TgCtWh6 + DFP group after taking DFP , the iron metabolism disorder was effectively improved , the IFN⁃γ content in the liver was significantly lowered (P < 0. 01) , and the inflammatory response of the liver was also reduced. Conclusion DFP can effectively improve iron metabolism disorders in the liver after infection with Toxoplasma gondii and reduce inflammatory damage in the liver.

It is difficult to maintain the initial posterior stability of the knee after posterior cruciate ligament reconstruction. Residual posterior knee laxity after operation is a problem of PCL reconstruction. It not only results in abnormal kinematics of the knee, but also leads to secondary meniscus injury and cartilage degeneration of the affected knee, and eventually leads to knee osteoarthritis, which may especially happen with persistent and severe posterior laxity. The main reasons of residual posterior knee laxity after PCL reconstruction are: improper treatment of the posterolateral corner injury, poor positioning of the femoral tunnel, small tibial slope, and unreasonable postoperative rehabilitation. There are some concepts and technologies, such as using artificial ligaments, tibial tunnel fixation with suspensory device or suspensory device combined with interference screws, enlargement of graft diameter, all-inside reconstruction combined suture augmentation, slow and gradual postoperative rehabilitation, which can eliminate or reduce the postoperative residual laxity, in order to improve clinical outcomes after PCL reconstruction. For the patients with flat tibial slopes, double-bundle PCL reconstruction and concurrent slope-increasing tibial osteotomy is suggested. It can reduce the risk of posterior laxity and improve the stability of the knee after operation.

The reconstruction of the posterior cruciate ligament (PCL) through the tibial tunnel is the most commonly used reconstruction technique after ligament injury.However, when the graft passes through the tibial tunnel back to the medial condyle of the femur, a sharp angle is formed at the proximal end of the tibia, called the "killer turn". The existence of the "killer turn" can lead to graft wear and expansion of adjacent tibial tunnel after PCL reconstruction, affecting the stability of the posterior knee joint after operation and even leading to the failure of operation. There are several techniques, such as modifified tibial tunnel technique which the proximal exit of tibial tunnel is located in the inferior and lateral aspect of the PCL tibial anatomic insertion site, increasing the angle between the tibial tunnel and the tibial plateau, creating a tibial tunnel from the anterior lateral side of the tibia, remnant preserving as soft tissue cushion, and inlay and onlay techniques for reconstructing PCL without using tibial tunnel reconstruction, can reduce the "killer turn" effect. The above 6 techniques, theoretically, can effectively reduce or eliminate the "killer turn" effect and improve the posterior stability of the knee joint after PCL reconstruction, so as to improve the clinical efficacy of PCL reconstruction. But, the number of cases using these techniques is relatively small, and their effectiveness, reliability, and advantages and disadvantages for patients still need more clinical practice to further explore and verify.

Objective:To clarify the clinicopathological, especially molecular, features of early-onset gastric cancer with the aim of informing analysis of treatment strategies.Methods:In this retrospective case-control study, we examined data from a dedicated gastric cancer database in Zhongshan Hospital affiliated to Fudan University. The original cohort comprised 2506 patients with gastric cancer who had undergone gastrectomy in Zhongshan Hospital Fudan University from July 2020 to October 2021, including 198 with early-onset gastric cancer (aged ≤45 years) and 2,308 with non-early gastric cancer. We used a simple random sampling method to select 396 of the 2,308 patients aged >45 years (ratio of 1:2) as the control group and then compared molecular diagnostic data and clinicopathological features of the two groups.Results:The median age was 39 years in the early-onset gastric cancer group, while 66 years in the control group. The clinicopathological features of early-onset gastric cancer included female predominance (59.1% [117/198] vs. 27.8% [110/396], χ 2=54.816, P<0.001), less comorbidity (32.3% [64/198] vs. 57.1% [226/396], χ 2=32.355, P<0.001), poorer differentiation (93.9% [186/198] vs. 74.5% [295/396], χ 2=30.777, P<0.001) and higher proportion of diffuse type (40.4% [80/198] vs. 15.9% [63/396], χ 2=69.639, P<0.001), distant metastasis (7.1% [14/198] vs. 2.8% [11/396], χ 2=6.034, P=0.014). Regarding treatment, distal gastrectomy was more commonly performed than proximal gastrectomy (55.1% [109/198] vs. 47.0% [186/396], 1.5% [3/198] vs. 8.3% [33/396], χ 2=11.644, P=0.003). Family history of gastric cancer, TNM stage, tumor size, lymph node dissection, nerve invasion, nodes harboring metastases, range of lymph node dissection, digestive tract reconstruction procedure, implementation of laparoscopic surgery, combined resection, and preoperative treatment did not differ significantly between the two groups (all P>0.05). Molecular diagnosis showed there was a smaller percentage of mismatch repair deficiency in the early-onset gastric cancer than in the control group (1.0% [2/198] vs. 10.1% [40/396], χ 2=16.301, P<0.001), and a higher rate of positivity for Claudin 18.2 (77.8% [154/198] vs. 53.0% [210/396], χ 2=5.442, P<0.001). HER-2 and Epstein–Barr virus positivity rates did not differ significantly between the two groups. Conclusion:Early-onset gastric cancer is a distinct type of gastric cancer with a high degree of malignancy, and treatment targeting Claudin 18.2 may be effective.

Objective:To clarify the clinicopathological, especially molecular, features of early-onset gastric cancer with the aim of informing analysis of treatment strategies.Methods:In this retrospective case-control study, we examined data from a dedicated gastric cancer database in Zhongshan Hospital affiliated to Fudan University. The original cohort comprised 2506 patients with gastric cancer who had undergone gastrectomy in Zhongshan Hospital Fudan University from July 2020 to October 2021, including 198 with early-onset gastric cancer (aged ≤45 years) and 2,308 with non-early gastric cancer. We used a simple random sampling method to select 396 of the 2,308 patients aged >45 years (ratio of 1:2) as the control group and then compared molecular diagnostic data and clinicopathological features of the two groups.Results:The median age was 39 years in the early-onset gastric cancer group, while 66 years in the control group. The clinicopathological features of early-onset gastric cancer included female predominance (59.1% [117/198] vs. 27.8% [110/396], χ 2=54.816, P<0.001), less comorbidity (32.3% [64/198] vs. 57.1% [226/396], χ 2=32.355, P<0.001), poorer differentiation (93.9% [186/198] vs. 74.5% [295/396], χ 2=30.777, P<0.001) and higher proportion of diffuse type (40.4% [80/198] vs. 15.9% [63/396], χ 2=69.639, P<0.001), distant metastasis (7.1% [14/198] vs. 2.8% [11/396], χ 2=6.034, P=0.014). Regarding treatment, distal gastrectomy was more commonly performed than proximal gastrectomy (55.1% [109/198] vs. 47.0% [186/396], 1.5% [3/198] vs. 8.3% [33/396], χ 2=11.644, P=0.003). Family history of gastric cancer, TNM stage, tumor size, lymph node dissection, nerve invasion, nodes harboring metastases, range of lymph node dissection, digestive tract reconstruction procedure, implementation of laparoscopic surgery, combined resection, and preoperative treatment did not differ significantly between the two groups (all P>0.05). Molecular diagnosis showed there was a smaller percentage of mismatch repair deficiency in the early-onset gastric cancer than in the control group (1.0% [2/198] vs. 10.1% [40/396], χ 2=16.301, P<0.001), and a higher rate of positivity for Claudin 18.2 (77.8% [154/198] vs. 53.0% [210/396], χ 2=5.442, P<0.001). HER-2 and Epstein–Barr virus positivity rates did not differ significantly between the two groups. Conclusion:Early-onset gastric cancer is a distinct type of gastric cancer with a high degree of malignancy, and treatment targeting Claudin 18.2 may be effective.

Objective : To observe the effects of TgCtwh3 and TgCtwh6, Toxoplasma gondii of Chinese 1 dominant genotype, on the ultrastructure of hippocampal of mice. Methods : Six⁃week⁃old female C57BL/6 mice were randomly divided into three groups: control group, TgCtwh3 infection group and TgCtwh6 infection group, with 6 mice in each group. Control group was intraperitoneally injected with sterile PBS. The TgCtwh3 infected group was intraperitoneally injected with 4 000 TgCtwh3 Toxoplasma gondii tachyzoites and fed for 6 days. The TgCtwh6 infected group was given 20 TgCtwh6 Toxoplasma gondii cysts by gavage and fed for 6 weeks. Then, all the animals were sacrificed, the brains were obtained for preparing electron microscope specimens and observed. Results : Compared with the control group, the cells in hippocampal of TgCtwh3 infected group showed nuclear shrinkage and necrosis; cytoplasmic and axon swelling; organelle disappearance; axonotmesis and other pathological changes. Compared with the control group, the cells in hippocampal of TgCtwh6 infected group showed swollen, vacuolated, crista⁃broken or membrane⁃broken mitochondria; lipofuscin deposition; dilation of golgi apparatus and other pathological changes. Conclusion Both TgCtwh3 and TgCtwh6 infected mice showed ultrastructural changes in hippocampal compared with normal mice. The TgCtwh3 group showed irreversible cell necrosis, while the TgCtwh6 group showed chronic cell damage mainly caused by mitochondrial structure destruction. These results lay a foundation to explore the pathogenesis of toxoplasmosis.