Objective:To investigate the expression levels, correlation, and diagnostic efficacy of peripheral blood CD8 +T lymphocyte subsets and different immune checkpoint markers in patients with Brucellosis. Method:A case-control study was conducted on 32 patients with acute phase brucellosis (27 males and 5 females, aged 36 (30, 43) years), 38 patients with chronic phase brucellosis (30 males and 8 females, aged 40 (32, 48) years), and 30 healthy controls (24 males and 6 females, aged 39 (32, 46) years), who underwent physical examination at Xinjiang Uygur Autonomous Region People′s Hospital from February 1, 2021 to December 31, 2023. All subjects had fasting blood sampling once in the morning. Flow cytometry was used to detect the proportion of peripheral blood lymphocyte subsets, the expression levels of CD8 +T lymphocyte surface programmed cell death receptor-1 (PD-1), T lymphocyte immunoglobulin receptor with Ig and ITIM domains (TIGIT), T cell immunoglobulin and mucin domain containing protein 3 (TIM-3), perforin and granzyme B. The changes in these indicators during the acute and chronic phases of the disease were observed, and correlation analysis was performed using Spearman′s method. Receiver Operating Characteristic Curve (ROC) analysis is used to evaluate the diagnostic value of immunological indicators with differences in acute and chronic brucellosis. Results:CD3 +T lymphocyte in the chronic group (70.71%±8.78%) is significantly lower than that in the healthy control group (74.65%±7.31%) ( P<0.05), and CD4 +T lymphocyte in the acute phase group (39.52%±5.85%) is also lower than that in the healthy control group (45.10%±5.18%) ( P<0.01); while CD8 +T lymphocyte in the acute group (31.73%±5.87%) is significantly higher than that in the chronic phase group (26.75%±4.71%) ( P<0.001). There was a statistically significant difference ( P<0.001) in CD8+CD28 -T lymphocyte among the acute group (69.85% (58.62%, 78.55%)), chronic group (86.46% (73.30%, 92.52%)) and healthy control group (25.39% (20.60%,32.90%)), when compared pairwise. The expression levels of immune checkpoint PD-1, TIGIT, and TIM-3 on the surface of CD8 +T lymphocytes were higher in both the acute and chronic groups than in the healthy control group ( P<0.001). The expression level of perforin secreted by CD8 +T lymphocytes in the acute and chronic groups was lower than that in the healthy control group ( P<0.05), while the expression level of granzyme B in the acute and chronic groups was higher than that in the healthy control group ( P<0.01). The proportion of CD8 +CD28 -T lymphocytes in brucellosis patients was positively correlated with the expression levels of TIGIT and TIM-3 ( r=0.624, 0.406, P<0.001). The ROC curve combined with the proportion of CD8 +CD28 -T lymphocytes and the proportion of TIGIT on the surface of CD8 +T lymphocytes can distinguish acute and chronic brucellosis. The Area Under Curve (AUC) is 0.973, which has certain implications for clinical differentiation of patients with acute and chronic diseases. Conclusion:The CD8 +T lymphocyte subsets in patients with brucellosis exhibit dynamic changes, accompanied by changes in relevant immune checkpoint molecules, and can regulate the activation and inhibition of the immune status of brucellosis patients. The synergistic effect of CD8 +CD28 -T cells and TIGIT/TIM-3 may be a key mechanism of driving chronicity, and their combined diagnosis can serve as a clinical staging marker.

[Objective] To analyze the loss rate of platelet donors and evaluate the strategies for establishing a platelet donor bank. [Methods] A total of 1 443 donors who joined the HLA and HPA gene donor bank for platelets in Henan Province from 2018 to 2020 were included in this study. Data on the total number of apheresis platelet donations, annual donation frequency, age at enrollment, donation habits (including the number of platelets donated per session and whether they had previously donated whole blood), and enrollment location were collected from the platelet donor information management system. Donor loss was determined based on the date of their last donation. The loss rates of different groups under various conditions were compared to assess the enrollment strategies. [Results] By the time the platelet bank was officially operational in 2022, 421 donors had been lost, resulting in an loss rate of 29% (421/1 443). By the end of 2023, the overall cumulative loss rate reached 52% (746/1 443). The loss rate was lower than the overall level in groups meeting any of the following conditions: total apheresis platelet donations exceeding 50, annual donation frequency of 10 or more, age at enrollment of 40 years or older, donation of more than a single therapeutic dose per session, or a history of whole blood donation two or more times. Additionally, loss rates varied across different enrollment locations, with higher enrollment numbers generally associated with higher loss rates. [Conclusion] Through a comprehensive analysis of donor loss, our center has adjusted its strategies for establishing the donor pool. These findings also provide valuable insights for other blood collection and supply institutions in building platelet donor banks.

Objective:To investigate the impact of para-tumoral micro-metastasis(PTMM) and other clinicopathological characteristics on the prognosis of patients with intrahepatic cholangiocarcinoma (ICC).Methods:This is a retrospective cohort study. Clinical data from 137 ICC patients who underwent radical resection at the Hepatobiliary Surgery Center, Department of General Surgery, Huashan Hospital, Fudan University, between January 2017 and December 2022 were analyzed retrospectively. The cohort included 91 males and 46 females, with age ( M(IQR)) of 63 (13) years (range: 32 to 82 years). Kaplan-Meier curves were used to estimate median survival times, while Log-rank tests assessed differences in overall survival (OS) and recurrence-free survival (RFS). Univariate and multivariate Cox regression models were employed to identify factors associated with OS and RFS. Results:The median OS for all 137 ICC patients was 34 months, with 1-, 2-, and 3-year OS rates of 90.7%, 69.4%, and 39.5%, respectively. The results of univariate and multivariate Cox analysis showed that Child-Pugh grade, CA19-9, carcinoembryonic antigen, and PTMM were independent prognostic factors for OS in ICC patients after radical resection (all P<0.05), Child-Pugh grade, maximum tumor diameter, whether lymph node metastasis, and PTMM were independent prognostic factors for RFS in radical resection in ICC patients (all P<0.05). PTMM-positive patients had a median OS of 21 months and median RFS of 12 months, whereas PTMM-negative patients exhibited a median OS exceeding 60 months and median RFS of 36 months. Log-rank tests demonstrated statistically significant differences in OS and RFS between PTMM-positive and PTMM-negative patients ( P<0.01 and P=0.001, respectively). Conclusion:Preliminary findings suggest that PTMM holds significant prognostic value in evaluating outcomes for ICC patients undergoing curative resection.

Objective:To investigate the expression levels, correlation, and diagnostic efficacy of peripheral blood CD8 +T lymphocyte subsets and different immune checkpoint markers in patients with Brucellosis. Method:A case-control study was conducted on 32 patients with acute phase brucellosis (27 males and 5 females, aged 36 (30, 43) years), 38 patients with chronic phase brucellosis (30 males and 8 females, aged 40 (32, 48) years), and 30 healthy controls (24 males and 6 females, aged 39 (32, 46) years), who underwent physical examination at Xinjiang Uygur Autonomous Region People′s Hospital from February 1, 2021 to December 31, 2023. All subjects had fasting blood sampling once in the morning. Flow cytometry was used to detect the proportion of peripheral blood lymphocyte subsets, the expression levels of CD8 +T lymphocyte surface programmed cell death receptor-1 (PD-1), T lymphocyte immunoglobulin receptor with Ig and ITIM domains (TIGIT), T cell immunoglobulin and mucin domain containing protein 3 (TIM-3), perforin and granzyme B. The changes in these indicators during the acute and chronic phases of the disease were observed, and correlation analysis was performed using Spearman′s method. Receiver Operating Characteristic Curve (ROC) analysis is used to evaluate the diagnostic value of immunological indicators with differences in acute and chronic brucellosis. Results:CD3 +T lymphocyte in the chronic group (70.71%±8.78%) is significantly lower than that in the healthy control group (74.65%±7.31%) ( P<0.05), and CD4 +T lymphocyte in the acute phase group (39.52%±5.85%) is also lower than that in the healthy control group (45.10%±5.18%) ( P<0.01); while CD8 +T lymphocyte in the acute group (31.73%±5.87%) is significantly higher than that in the chronic phase group (26.75%±4.71%) ( P<0.001). There was a statistically significant difference ( P<0.001) in CD8+CD28 -T lymphocyte among the acute group (69.85% (58.62%, 78.55%)), chronic group (86.46% (73.30%, 92.52%)) and healthy control group (25.39% (20.60%,32.90%)), when compared pairwise. The expression levels of immune checkpoint PD-1, TIGIT, and TIM-3 on the surface of CD8 +T lymphocytes were higher in both the acute and chronic groups than in the healthy control group ( P<0.001). The expression level of perforin secreted by CD8 +T lymphocytes in the acute and chronic groups was lower than that in the healthy control group ( P<0.05), while the expression level of granzyme B in the acute and chronic groups was higher than that in the healthy control group ( P<0.01). The proportion of CD8 +CD28 -T lymphocytes in brucellosis patients was positively correlated with the expression levels of TIGIT and TIM-3 ( r=0.624, 0.406, P<0.001). The ROC curve combined with the proportion of CD8 +CD28 -T lymphocytes and the proportion of TIGIT on the surface of CD8 +T lymphocytes can distinguish acute and chronic brucellosis. The Area Under Curve (AUC) is 0.973, which has certain implications for clinical differentiation of patients with acute and chronic diseases. Conclusion:The CD8 +T lymphocyte subsets in patients with brucellosis exhibit dynamic changes, accompanied by changes in relevant immune checkpoint molecules, and can regulate the activation and inhibition of the immune status of brucellosis patients. The synergistic effect of CD8 +CD28 -T cells and TIGIT/TIM-3 may be a key mechanism of driving chronicity, and their combined diagnosis can serve as a clinical staging marker.

Objective:To investigate the impact of para-tumoral micro-metastasis(PTMM) and other clinicopathological characteristics on the prognosis of patients with intrahepatic cholangiocarcinoma (ICC).Methods:This is a retrospective cohort study. Clinical data from 137 ICC patients who underwent radical resection at the Hepatobiliary Surgery Center, Department of General Surgery, Huashan Hospital, Fudan University, between January 2017 and December 2022 were analyzed retrospectively. The cohort included 91 males and 46 females, with age ( M(IQR)) of 63 (13) years (range: 32 to 82 years). Kaplan-Meier curves were used to estimate median survival times, while Log-rank tests assessed differences in overall survival (OS) and recurrence-free survival (RFS). Univariate and multivariate Cox regression models were employed to identify factors associated with OS and RFS. Results:The median OS for all 137 ICC patients was 34 months, with 1-, 2-, and 3-year OS rates of 90.7%, 69.4%, and 39.5%, respectively. The results of univariate and multivariate Cox analysis showed that Child-Pugh grade, CA19-9, carcinoembryonic antigen, and PTMM were independent prognostic factors for OS in ICC patients after radical resection (all P<0.05), Child-Pugh grade, maximum tumor diameter, whether lymph node metastasis, and PTMM were independent prognostic factors for RFS in radical resection in ICC patients (all P<0.05). PTMM-positive patients had a median OS of 21 months and median RFS of 12 months, whereas PTMM-negative patients exhibited a median OS exceeding 60 months and median RFS of 36 months. Log-rank tests demonstrated statistically significant differences in OS and RFS between PTMM-positive and PTMM-negative patients ( P<0.01 and P=0.001, respectively). Conclusion:Preliminary findings suggest that PTMM holds significant prognostic value in evaluating outcomes for ICC patients undergoing curative resection.

Objective To evaluate the clinical value of neutrophil CD64(nCD64)index as a novel biomark-er in the differential diagnosis of acute and chronic brucellosis.Methods A total of 38 patients with acute bru-cellosis and 48 patients with chronic brucellosis diagnosed in the People's Hospital of Xinjiang Uygur Autono-mous Region from February 2021 to July 2023 were included.Peripheral blood of the patients was collected and nCD64 index was detected by flow cytometry,and the correlation between nCD64 index and disease severi-ty was analyzed.Receiver operating characteristic(ROC)curve was used to analyze the sensitivity and the specificity of nCD64 index in differentially diagnosing acute and chronic brucellosis.Meanwhile,Rose-Bengal Plate Test(RBPT)and Standard Tube Agglutination Test(SAT)were used as controls to evaluate the clini-cal diagnostic value of the three.Results The nCD64 index of acute brucellosis patients was higher than that of chronic brucellosis patients(U=216.00,P<0.001),and the index was positively correlated with the sever-ity of the disease(r=0.670,P<0.001).The ROC curve analysis results showed that the area under the curve of nCD64 index in the differential diagnosis of acute and chronic brucellosis was 0.882(95%CI:0.811-0.952,P<0.001),the cut-off value was 2.81,and sensitivity,specificity,positive predictive value,negative predictive value and accuracy were 83.3%,81.6%,80.4%,81.9%and 82.6%,respectively.The efficacy of nCD64 index in differential diagnosis of nCD64 index was significantly better than those of the qualitative tests of RBPT and SAT.Conclusion nCD64 index has favourable sensitivity and specificity in the differential diag-nosis of acute and chronic brucellosis,and tends to reflect the severity of the disease.It has clinical value in the differential diagnosis of acute brucellosis and chronic brucellosis,and plays an important role in the early diag-nosis and treatment effect monitoring of brucellosis.

Objective To investigate the characteristics of changes in interleukin(IL)-33 and regulatory T(Treg)cells in brucellosis,to verify the regulatory effect of IL-33 on Treg cells,so as to clarify the immune mechanism of IL-33 on Treg cells in brucellosis.Methods The peripheral blood of 39 patients with brucellosis treated in the People's Hospital of Xinjiang Uygur Autonomous Region from January to December 2021(the brucellosis group)and 42 healthy controls(the healthy control group)who underwent physical examination during the same period were collected.The serum IL-33 level was detected by AimPlex kit,and the proportion of Treg cells was detected by flow cytometry.Peripheral blood mononuclear cell(PBMC)was extracted and cultured in vitro to observe the proportion and mRNA expression levels of forkhead box protein P3(Foxp3)after stimulation and blocking of IL-33.Results Compared with the healthy control group,the level of IL-33 and the proportion of Treg cells in brucellosis group were significantly increased,with statistical significance(P＜0.05).In vitro tests showed that the Foxp3 proportion and mRNA expression level of PBMC in the two groups were significantly increased after IL-33 stimulation,and significantly decreased after IL-33 blocking,with statistical significance(P＜0.001).Conclusion IL-33 and Treg cells increased significantly in brucellosis patients,and IL-33 promoted the immune function of Treg cells.Blocking IL-33 is expected to be a potential target for immunotherapy of brucellosis.

OBJECTIVE: To design and construct a graphene oxide (GO)/silver nitrate (Ag₃PO₄)/chitosan (CS) composite coating for rapidly killing bacteria and preventing postoperative infection in implant surgery. METHODS: GO/Ag₃PO₄ composites were prepared by ion exchange method, and CS and GO/Ag₃PO₄ composites were deposited on medical titanium (Ti) sheets successively. The morphology, physical image, photothermal and photocatalytic ability, antibacterial ability, and adhesion to the matrix of the materials were characterized. RESULTS: The GO/Ag₃PO₄ composites were successfully prepared by ion exchange method and the heterogeneous structure of GO/Ag₃PO₄ was proved by morphology phase test. The heterogeneous structure formed by Ag₃PO₄ and GO reduced the band gap from 1.79 eV to 1.39 eV which could be excited by 808 nm near-infrared light. The photothermal and photocatalytic experiments proved that the GO/Ag₃PO₄/CS coating had excellent photothermal and photodynamic properties. In vitro antibacterial experiments showed that the antibacterial rate of the GO/Ag₃PO₄/CS composite coating against Staphylococcus aureus reached 99.81% after 20 minutes irradiation with 808 nm near-infrared light. At the same time, the composite coating had excellent light stability, which could provide stable and sustained antibacterial effect. CONCLUSION GO/Ag₃PO₄/CS coating can be excited by 808 nm near infrared light to produce reactive oxygen species, which has excellent antibacterial activity under light.
Chitosan ; Silver Nitrate ; Titanium ; Anti-Bacterial Agents/pharmacology* ; Coloring Agents

Bispecific antibody (BsAbs) are antibodies (Abs) containing two different antigen-binding sites in one molecule. In the last decade, three BsAbs drugs have been approved for therapeutic use. Meanwhile there are a number of BsAbs in preclinical or clinical studies. In this review, we describe BsAb design, discovery, mechanism of action, and the recent research progress in developing BsAbs.

Objective To study the clinical features and risk factors of acute pulmonary reperfusion injury after operation in neonates with severe pulmonary stenosis or pulmonary atresia.Method From February 2014 to February 2018,a retrospective analysis was performed in patients with critical pulmonary stenosis or pulmonary atresia who received percutaneous balloon pulmonary valvuloplasty (PBPV) in the neonatal intensive care unit of our hospital.Clinical characteristics,perioperative cardiac structure,hemodynamic data and biochemical results were collected.The neonates were assigned into injury group if they had acute lung reperfusion injury,and non-injury group if not.The risk factors of acute lung reperfusion injury were analyzed using multi-variate Logistic regression model.Result A total of 32 patients (24 prenatal diagnosis and 8 postnatal diagnosis) with severe pulmonary stenosis or pulmonary atresia with intact ventricular septum were enrolled.The main manifestations were dyspnea and cyanosis.Intravenous prostaglandin E was administered to keep the ductus arteriosus open.The age of operation ranged from 1 to 52 days and the median age was 7.5 days.Postoperative acute lung reperfusion injury occurred in 7 cases (21.9％).Preoperative and intraoperative pulmonary valve annulus diameter,balloon diameter,preoperative hemoglobin,hematocrit and blood albumin were significantly lower in the injury group.The operation duration,total length of hospital stay and postoperative duration were longer than in the non-injury group,the differences were statistically significant (P ＜ 0.05).Multi-variate Logistic regression analysis showed that the diameter of pulmonary valve annulus (OR =5.814,95％CI 1.106 ～30.568),preoperative blood albumin (OR =1.361,95％ CI 1.063 ～ 1.742),and hematocrit (OR =1.173,95％ CI 1.010 ～ 1.363) were risk factors of acute lung reperfusion injury,with statistically significant differences (P ＜ 0.05).Conclusion Acute lung reperfusion injury is one of the common complications after the operation of severe pulmonary stenosis or pulmonary atresia.The severity of pulmonary valve annulus stenosis,preoperative hematocrit and blood albumin level may be the risk factors of postoperative acute lung reperfusion injury.