Objective:To investigate the potential diagnostic value of peripheral blood soluble programmed cell death protein 1 (sPD-1) and soluble programmed cell death protein ligand 1 (sPD-L1) in acute rejection (AR) following liver transplantation using a rat model.Methods:A rat liver transplantation AR model was established, with the AR group (Lewis→BN) set as the experimental group (n=6) and the non-AR group (BN→BN) as the control group (n=6). Peripheral blood sPD-1 and sPD-L1 concentrations were measured using enzyme-linked immunosorbent assay (ELISA) at 1 day before transplantation and at 1, 3, and 7 days postoperatively. On postoperative day 7, the expression levels of PD-1 and PD-L1 in liver tissues were detected by immunohistochemistry (IHC). Independent samples t-test and repeated measures ANOVA were used to compare the results between the two groups. Pearson correlation analysis was conducted to evaluate the relationship between sPD-1, sPD-L1, the sPD-1/sPD-L1 ratio, and the rejection activity index.Results:On postoperative day 7, the experimental group exhibited significantly higher peripheral sPD-1 levels (218.59±36.88 vs. 164.95±15.82 ng/L) and a higher sPD-1/sPD-L1 ratio (0.44±0.12 vs. 0.36±0.07), but lower sPD-L1 levels (379.56±73.41 vs. 423.64±96.55 ng/L) compared to the control group (all P<0.05). Correlation analysis revealed a significant positive correlation between sPD-1 levels and the rejection activity index ( r=0.680, P<0.05), as well as between the sPD-1/sPD-L1 ratio and the rejection activity index ( r=0.795, P<0.01), while no correlation was observed between sPD-L1 levels and the rejection activity index. IHC demonstrated positive PD-1 and PD-L1 expression in the liver tissues of the experimental group, whereas the control group showed negative expression. Conclusion:Peripheral blood sPD-1 levels and the sPD-1/sPD-L1 ratio are significantly associated with AR after liver transplantation in rats, suggesting their potential as biomarkers for diagnosing AR in liver transplant recipients.

Objective:To investigate the potential diagnostic value of peripheral blood soluble programmed cell death protein 1 (sPD-1) and soluble programmed cell death protein ligand 1 (sPD-L1) in acute rejection (AR) following liver transplantation using a rat model.Methods:A rat liver transplantation AR model was established, with the AR group (Lewis→BN) set as the experimental group (n=6) and the non-AR group (BN→BN) as the control group (n=6). Peripheral blood sPD-1 and sPD-L1 concentrations were measured using enzyme-linked immunosorbent assay (ELISA) at 1 day before transplantation and at 1, 3, and 7 days postoperatively. On postoperative day 7, the expression levels of PD-1 and PD-L1 in liver tissues were detected by immunohistochemistry (IHC). Independent samples t-test and repeated measures ANOVA were used to compare the results between the two groups. Pearson correlation analysis was conducted to evaluate the relationship between sPD-1, sPD-L1, the sPD-1/sPD-L1 ratio, and the rejection activity index.Results:On postoperative day 7, the experimental group exhibited significantly higher peripheral sPD-1 levels (218.59±36.88 vs. 164.95±15.82 ng/L) and a higher sPD-1/sPD-L1 ratio (0.44±0.12 vs. 0.36±0.07), but lower sPD-L1 levels (379.56±73.41 vs. 423.64±96.55 ng/L) compared to the control group (all P<0.05). Correlation analysis revealed a significant positive correlation between sPD-1 levels and the rejection activity index ( r=0.680, P<0.05), as well as between the sPD-1/sPD-L1 ratio and the rejection activity index ( r=0.795, P<0.01), while no correlation was observed between sPD-L1 levels and the rejection activity index. IHC demonstrated positive PD-1 and PD-L1 expression in the liver tissues of the experimental group, whereas the control group showed negative expression. Conclusion:Peripheral blood sPD-1 levels and the sPD-1/sPD-L1 ratio are significantly associated with AR after liver transplantation in rats, suggesting their potential as biomarkers for diagnosing AR in liver transplant recipients.

BACKGROUND AND AIM: Remnant cholesterol (remnant-C) mediates the progression of major adverse cardiovascular events. It is unclear whether remnant-C, and particularly cumulative exposure to remnant-C, is associated with nonalcoholic fatty liver disease (NAFLD). This study aimed to explore whether remnant-C, not only baseline but cumulative exposure, can be used to independently evaluate the risk of NAFLD. METHODS: This study included 1 cohort totaling 21,958 subjects without NAFLD at baseline who underwent at least 2 repeated health checkups and 1 sub-cohort totaling 2,649 subjects restricted to those individuals with at least 4 examinations and no history of NAFLD until Exam 3. Cumulative remnant-C was calculated as a timeweighted model for each examination multiplied by the time between the 2 examinations divided the whole duration. Cox regression models were performed to estimate the association between baseline and cumulative exposure to remnant-C and incident NAFLD. RESULTS: After multivariable adjustment, compared with the quintile 1 of baseline remnant-C, individuals with higher quintiles demonstrated significantly higher risks for NAFLD (hazard ratio [HR] 1.48, 95%CI 1.31-1.67 for quintile 2; HR 2.07, 95%CI 1.85-2.33 for quintile 3; HR 2.55, 95%CI 2.27-2.88 for quintile 4). Similarly, high cumulative remnant-C quintiles were significantly associated with higher risks for NAFLD (HR 3.43, 95%CI 1.95-6.05 for quintile 2; HR 4.25, 95%CI 2.44-7.40 for quintile 3; HR 6.29, 95%CI 3.59-10.99 for quintile 4), compared with the quintile 1. CONCLUSION Elevated levels of baseline and cumulative remnant-C were independently associated with incident NAFLD. Monitoring immediate levels and longitudinal trends of remnant-C may need to be emphasized in adults as part of NAFLD prevention strategy.
Adult ; Humans ; Cohort Studies ; Non-alcoholic Fatty Liver Disease/etiology* ; Cholesterol ; Proportional Hazards Models ; Risk Factors

Objective:To explore the effects of acupuncture combined with Buyang Huanwu Decoction on intestinal flora in cerebral blood flow hypo perfusion model rats with carotid artery stenosis.Methods:Totally 40 rats were randomly divided into sham-operation group, model group, TCM treatment group and acupuncture and drug combination treatment group, with 10 rats in each group. Except the sham-operation group, the other groups were prepared cerebral ischemia model by needle control and thread embolism method. TCM treatment group received Buyang Huanwu Decoction 100 mg/kg for gavage, once a day, and the intervention lasted for 2 weeks. In the acupuncture and drug combination group, based on the TCM treatment group, Baihui and its left and right sides of 2 mm were selected for acupuncture, once a day, and continuous intervention was performed for 2 weeks. Neurological function evaluation and behavioral function score were performed 7 and 14 days after administration, respectively. 16S rRNA sequencing was used to comprehensively characterize the structure and composition of fecal microflora of rats in each group. Linear discriminant analysis Effect Size (LEfSe) was used to analyze the difference of intestinal bacteria among groups.Result:On the 7th and 14th day after administration, compared with the model group, the neurological function score in the TCM treatment group and the acupuncture and drug combination group decreased ( P<0.05), and the behavioral function score increased ( P<0.05). Compared with model group, the Shannon index of TCM treatment group and acupuncture and drug combination group increased ( P<0.05). The abundance of Firmicutes increased ( P<0.05), and the abundance of Bacteroidetes and Proteobacteria decreased ( P<0.05); the abundance of Clostridia increased ( P<0.05), and the abundance of Gammaproteobacteria decreased ( P<0.05). The abundance of Escherichia-Shigella and Bacteroides decreased ( P<0.05); the abundance of lactobacillus significantly increased ( P<0.05). Conclusion:Acupuncture combined with Buyang Huanwu Decoction can improve the symptoms of cerebral hypoperfusion model rats with carotid artery stenosis, and the mechanism may be to increase the abundance of probiotics.

Objective:To establish and validate a fluorescence PCR with internal positive control for rapid Mycoplasma detection. Methods:A fluorescence PCR with internal positive control for Mycoplasma detection was developed and verified for its specificity, limit of detection, and robustness. A sample of fever with thrombocytopenia syndrome (SFTSV) virus strains was tested with this method, and the result was compared with those of culture method and indicator cell culture method. Results:The established fluorescence PCR had good specificity and could amplify 11 kinds of plasmids containing Mycoplasma 16S rRNA gene with high efficiency. There was no cross reaction with the genomic DNAs of Clostridium sporogenes, Clostridium acetobutylicum, Lactobacillus acidophilus, Streptococcus pneumoniae, Bacillus subtilis, Staphylococcus aureus, Salmonella enteritidis, Escherichia coli, Pseudomonas aeruginosa, Aspergillus niger, Candida albicans, Vero cells, RD cells, and SF9 cells. The amplification efficiency of the internal positive control was basically consistent with that of the target gene of Mycoplasma, suggesting that the internal positive control could be used to detect the presence of PCR inhibitors. The sensitivity of the established method was high, and the detection limit was 10 colony-forming unit (CFU)/ml for Mycoplasma fermentans, 5 CFU/ml for Mycoplasma arginine, 5 CFU/ml for Mycoplasma gallisepticum, 5 CFU/ml for Mycoplasma hyorhinis, 5 CFU/ml for Acholeplasma laidlawii, 5 CFU/ml for Mycoplasma orale, 5 CFU/ml for Mycoplasma pneumoniae, 5 CFU/ml for Mycoplasma synoviae, and 1 CFU/ml for Spiroplasma citri by 7500 Fast real-time PCR system. At the detection limit of each species, there was no significant difference in the positive detection rate using different thermal cycler types. The established fluorescence PCR, culture method, and indicator cell culture were performed to detect Mycoplasma in the sample of SFTSV virus strains, and the results all showed Mycoplasma contamination. Conclusions:The established fluorescence PCR has high specificity, sensitivity, and robustness, and can be used as an alternative method for rapid detection of Mycoplasma.

Objective:To explore the feasibility of the maze design of a proton therapy system.Methods:According to the construction scheme of a medical proton therapy system, the ambient neutron dose equivalent rate outside the maze entrance was estimated by numerical analytical formula, and compared with the monitoring result.Results:The ambient neutron dose equivalent rate was estimated at between 3.85×10 -2 μSv/h and 7.57 μSv/h and measured to be ≤1.56 μSv/h. Conclusions:The maze entrance was designed to have 5 sections of tunnel for cyclotron room and 3 sections of tunnel for treatment room, respectively, each with designated length and width. The levels of ambient dose equivalent rate outside the maze entrance is acceptable.

Osteofacial compartment syndrome (OCS) is one of the serious complications in traumatic orthopedics. If not treated in time, OCS may result in irreversible damage to nerve and muscle,even amputation or death in serious condition. 5P presents to be the classic clinical diagnosis of OCS, but it is highly subjective and cannot timely and accurately judge the progression of the disease. Intracompartment pressure manometry is the main auxiliary method for the diagnosis of OCS. Although there are many manometry methods, there is still no authoritative pressure threshold as the diagnosis standard. Clinicians often aggressively perform fasciotomy to avoid serious complications, leading to unnecessary fasciotomy. The authors retrospectively reviewed the data of patients with OCS treated at Air Force Hospital of Eastern Theater of PLA from March 2010 to March 2020 and found that some patients with OCS had gradual alleviation of clinical symptoms after appropriate conservative treatments such as brace releasing, limb stabilization and swelling subsidence, with no need of fasciotomy. However, the symptoms of some patients progressively aggravated after the above-mentioned traditional treatments and timely fasciotomy was required. The authors graded the severity of OCS and proposed for the first time the OCS grading criteria according to quantitative clinical results and quantitative indicators such as ratio of mean blood flow velocity of bilateral arteries and pulse wave changes, aiming to take corresponding intervention measures for patients with different OCS classifications, carry out more precise treatment and avoid unnecessary fasciotomy.

The aim of this study was to identify whether periodontitis induces gut microbiota dysbiosis via invasion by salivary microbes. First, faecal and salivary samples were collected from periodontally healthy participants (PH group, n = 16) and patients with severe periodontitis (SP group, n = 21) and analysed by 16S ribosomal RNA sequencing. Significant differences were observed in both the faecal and salivary microbiota between the PH and SP groups. Notably, more saliva-sourced microbes were observed in the faecal samples of the SP group. Then, the remaining salivary microbes were transplanted into C57BL6/J mice (the C-PH group and the C-SP group), and it was found that the composition of the gut microbiota of the C-SP group was significantly different from that of the C-PH group, with Porphyromonadaceae and Fusobacterium being significantly enriched in the C-SP group. In the colon, the C-SP group showed significantly reduced crypt depth and zonula occludens-1 expression. The mRNA expression levels of pro-inflammatory cytokines, chemokines and tight junction proteins were significantly higher in the C-SP group. To further investigate whether salivary bacteria could persist in the intestine, the salivary microbiota was stained with carboxyfluorescein diacetate succinimidyl ester and transplanted into mice. We found that salivary microbes from both the PH group and the SP group could persist in the gut for at least 24 h. Thus, our data demonstrate that periodontitis may induce gut microbiota dysbiosis through the influx of salivary microbes.
Animals ; Dysbiosis ; Gastrointestinal Microbiome ; Humans ; Mice ; Mice, Inbred C57BL ; Microbiota ; Periodontitis ; RNA, Ribosomal, 16S/metabolism*

Osteoarthritis (OA), in which M1 macrophage polarization in the synovium exacerbates disease progression, is a major cause of cartilage degeneration and functional disabilities. Therapeutic strategies of OA designed to interfere with the polarization of macrophages have rarely been reported. Here, we report that SHP099, as an allosteric inhibitor of src-homology 2-containing protein tyrosine phosphatase 2 (SHP2), attenuated osteoarthritis progression by inhibiting M1 macrophage polarization. We demonstrated that M1 macrophage polarization was accompanied by the overexpression of SHP2 in the synovial tissues of OA patients and OA model mice. Compared to wild-type (WT) mice, myeloid lineage conditional Shp2 knockout (cKO) mice showed decreased M1 macrophage polarization and attenuated severity of synovitis, an elevated expression of cartilage phenotype protein collagen II (COL2), and a decreased expression of cartilage degradation markers collagen X (COL10) and matrix metalloproteinase 3 (MMP3) in OA cartilage. Further mechanistic analysis showed thatSHP099 inhibited lipopolysaccharide (LPS)-induced Toll-like receptor (TLR) signaling mediated by nuclear factor kappa B (NF-κB) and PI3K-AKT signaling. Moreover, intra-articular injection of SHP099 also significantly attenuated OA progression, including joint synovitis and cartilage damage. These results indicated that allosteric inhibition of SHP2 might be a promising therapeutic strategy for the treatment of OA.

Objective:To investigate the etiology,clinical features and prognosis of pediatric liver retransplantation.Methods:The data of 1 024 cases of pediatric liver transplantation (<18 years old) from January 2014 to December 2019 operated at Tianjin First Central Hospital were collected,retrospectively. Retransplantation was performed in 26 cases,among which 25 cases received secondary liver transplantation and 1 case received a third liver transplantation. There were 13 male and 12 female patients among the 25 patients. The median age was 12.9(20.5) months(range: 5.8 to 134.8 months), the body weight was 8.0(5.6) kg(range: 5.0 to 30.0 kg) at the time of retransplantation. The pediatric end-stage liver disease(PELD) score was 17.0(21.3) (range: 0 to 45) before retransplantation. The etiology of retransplantation was biliary complications in 7 cases,primary nonfunction of liver graft in 5 cases,antibody-mediated rejection in 4 cases,hepatic artery thrombosis in 3 cases,portal vein thrombosis in 3 cases,concomitant hepatic artery and portal vein thrombosis in 2 cases,thrombogenesis of inferior Vena Cava in 1 case and sinusoidal obstruction syndrome in 1 case. The patients were divided into two groups according to the time interval(30 days) between two liver transplantations,8 patients were classified into early-retransplantation(≤30 days) group and 18 patients were classified into late-retransplantation (>30 days) group. The etiology of liver retransplantation,pre-transplant score,time interval between two transplantations,surgical aspects,major complications and survival rates were compared between the two groups. Continuous variables with normal distribution were compared with t test,while Mann-Whitney U test was applied to compare variables without normal distribution. Categorical variables were compared with chi-square test. The survival curves were created by Kaplan-Meier method and compared by Log Rank test. Results:The median follow-up time was 26.8(30.2) months(range: 1 day to 85.7 months), and the incidence of retransplantation was 1.9%. In the early-retransplantation group,the duration of surgery was (439.8±151.0)minutes,the graft-to-recipient weight ratio was 5.0(1.8)%(range:3.6% to 6.1%),the main cause for retransplantation were primary nonfunction and vascular complications. In the late-retransplantation group,the duration of surgery was (604.4±158.0)minutes,the graft-to-recipient weight ratio was 3.4(2.1)%(range:1.4% to 5.3%),the main cause for retransplantation were biliary complications,antibody mediated rejection and vascular complications.The 3-month,1-year and 2-year recipient survival rates in the early-retransplantation group were all 62.3%,while the recipient survival rates in the late-retransplantation group were 100%,93.8% and 93.8%,respectively. The difference of recipient survival rates was significant between the early-retransplantation group and the late-retransplantation group( P=0.019). The overall 3-month,1-year and 3-year recipient survival rates after the primary liver transplantation were 97.1%,95.4%,94.1%,respectively. Conclusions:The vascular complications,biliary complications,primary nonfunction and antibody-mediated rejection are the main causes of liver retransplantation.The PELD score is higher in patients receiving early retransplantation,while the surgery is relatively more complex in patients receiving late retransplantation,which is reflected by longer duration of surgeries. Patients in the late-retransplantation group showed similar recipient survival rates with primary liver transplantation recipients,and the survival rates are superior to those of patients in the early-retransplantation group. Infection and multiple organ failure are the most common fatal causes after retransplantation.