Objective: To investigate the incidence of occult hepatitis B virus infection (OBI) in spouses of asymptomatic hepatitis B virus (HBV) infected individuals in Shenzhen, China, and to analyze their serological and molecular characteristics and possible transmission routes, so as to propose refined strategies for blood safety. Methods: After rapid screening for HBsAg at the blood collection sites, samples from HBsAg-positive blood donors and their concurrently donating spouses were collected. All samples were tested for hepatitis B serological markers by electrochemiluminescence immunoassay (ECLI). Simultaneously, HBV nucleic acid extractiona, nested PCR amplification, gene sequencing of S and BCP/PC regions and qPCR were conducted. Results: A total of 112 samples were collected, including 56 from HBsAg positive donors and 56 from their spouses. All donors were confirmed as HBsAg+/DNA+/anti-HBc+, indicative of asymptomatic chronic hepatitis (CHB) infection. Among their 56 spouses, 11 (19.6%) were identified as HBV DNA+. The prevalence was higher in males (23.1%) than in females (16.7%). Six spouses (10.8%) had OBI, three of whom (5.4%) were negative in routine blood screening tests. The residual risk of HBV were estimated as 1∶127 (95%CI, 1∶356 to 1∶66). Among infected couples, immune escape mutation (E164D) and glycosylation mutations (I126T and T131N/M133T) were identified. Furthermore, sequence analysis suggested partner-to-partner transmission in eight cases. Conclusion: A substantial proportion (19.6%) of spouses of asymptomatic HBV infected donors were HBV-positve, with an OBI prevalence of 10.9%. Among these, 5.4% were negative in routine tests. To ensure blood safety, we recommend that spouses of HBV infected individuals be deferred from blood donation.

Objective: Data on syncopal donation-related vasovagal reaction (DRVR) collected from 74 blood centers between 2020 and 2023 was statistically analyzed to provide a reference for developing preventive strategies against syncopal DRVR. Methods: Data on blood donation adverse reactions and basic information of donors from 2020 to 2023 were collected through the information management system at monitoring sentinel sites. Statistical analysis was performed on the following aspects of syncopal DRVR: characteristics of donors who experienced syncope, reported incidence, triggers, duration, presence and occurrence time of syncope-related trauma, clinical management including outpatient and inpatient treatment, and severity grading. Results: From 2020 to 2023, 45 966 donation-related adverse reactions were recorded. Of these, 1 665 (3.72%) cases were syncopal DRVR. The incidence of syncopal DRVR decreased with age, being the highest in the 18-22 age group. Incidence was significantly higher in female donors than male donors, in first-time donors than repeat donors, and in university and individual donors than group donors (all P<0.05). There was no statistically significant difference among different blood donation locations (P>0.05). The top three triggers were tension, fatigue, and needle phobia or fear of blood. Among syncopal DRVR cases, 60.36% occurred during blood collection, 87.63% lasted for less than 60 seconds, and 5.05% were accompanied by trauma. Notably, 57.14% of these traumas occurred after donor had left the blood collection site. Syncope severity was graded based on required treatment: grade 1 (fully recovered without treatment, 95.50%); grade 2 (recovered after outpatient treatment, 4.02%); and grade 3 (recovered after inpatient treatment, 0.48%). Conclusion: By analyzing the data of syncopal DRVR cases, it is possible to provide a reference for formulating blood donor safety policies.

Objective: To retrospectively analyze the prevalence of hepatitis C virus (HCV) infection among voluntary blood donors in Qinghai Province over a ten-year period and to provide evidence for refining blood safety screening strategies. Methods: Blood samples from 362 066 blood donors in Qinghai collected between January 2015 and April 2024 were simultaneously screened using enzyme-linked immunosorbent assay (ELISA) and nucleic acid testing (NAT). Follow-up was conducted for donors with reactive HCV RNA screening results, and alanine transaminase (ALT) was detected by rate method. Results: The HCV positive rate among blood donors in Qinghai was 0.22%. Gender, marital status, number of blood donations, and educational level were associated with HCV infection. Significant differences in HCV positive rates were observed among donors across regions and ethnic groups. The HCV positive rate among donors in Golog Tibetan Autonomous Prefecture (with an average altitude of 4 330 m) was significantly higher than that in Xining (0.52% vs 0.21%, P<0.001). Positivity rates were also significantly higher in Salar (0.84%), Hui (0.81%), Zang (0.60%), and Tu (0.45%) ethnic groups compared to the Han ethnic group (0.17%) (P<0.001). The abnormal rate of ALT in HCV-positive donors was higher than in non-HCV donors (6.13% vs 1.55%) (P<0.001). Conclusion: The relatively high HCV positivity rate among blood donors in Qinghai highlights the need for further investigation into viral sources, risk factors, and transmission routes. Optimized screening strategies are essential to ensure blood safety.

[Objective] To investigate the factors associated with alanine aminotransferase (ALT) abnormalities in multi-ethnic blood donors across five Zang autonomous prefectures in the plateau regions of Qinghai Province, and to provide evidence for ensuring blood safety and formulating screening strategies. [Methods] A retrospective analysis was performed on the ALT abnormal test results of blood donors in the Zang autonomous prefectures of Qinghai from 2022 to 2024. The correlations between ALT levels and factors including gender, age, altitude, and infectious markers were investigated. [Results] The overall ALT unqualified rate among blood donors in this region was 9.01%. Significant differences in ALT levels were observed across genders and age groups (P<0.05). Variations in ALT abnormality rates were also noted among different plateau regions (P<0.05). Overall, ALT values exhibited an increasing trend with rising altitude. The average ALT unqualified rates were 11.19% in Zang donors, 7.96% in Han donors, and 4.79% in donors from other ethnic groups (P<0.05). No statistically significant association was observed between ALT abnormality and the presence of HBV/HCV infectious markers (P>0.05). [Conclusion] In the plateau areas of Qinghai, multi-ethnic blood donors have a relatively high ALT levels and ALT unqualified rates, showing distinct regional characteristics. ALT elevation in voluntary blood donors is related to non-pathological factors such as gender, age, and dietary habits, but not to infectious indicators.

Objective To explore the application of plasma 7 microRNA (miR7) testing in the differential diagnosis of very early-stage hepatocellular carcinoma (HCC). Methods This study is a multicenter real-world study. Patients with single hepatic lesion (maximum diameter≤2 cm) who underwent plasma miR7 testing at Zhongshan Hospital, Fudan University, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Anhui Provincial Hospital, and Peking University People’s Hospital between January 2019 and December 2024 were retrospectively enrolled. Patients were divided into very early-stage HCC group and non-HCC group, and the clinical pathological characteristics of the two groups were compared. The value of plasma miR7 levels, alpha-fetoprotein (AFP), and des-gamma-carboxy prothrombin (DCP) in the differential diagnosis of very early-stage HCC was evaluated using receiver operating characteristic (ROC) curves and area under the curve (AUC). In patients with both negative AFP and DCP (AFP＜20 ng/mL, DCP＜40 mAU/mL), the diagnostic value of plasma miR7 for very early-stage HCC was analyzed. Results A total of 64 528 patients from 4 hospitals underwent miR7 testing, and 1 682 were finally included, of which 1 073 were diagnosed with very early-stage HCC and 609 were diagnosed with non-HCC. The positive rate of miR7 in HCC patients was significantly higher than that in non-HCC patients (67.9% vs 24.3%, P＜0.001). ROC curves showed that the AUCs for miR7, AFP, and DCP in distinguishing HCC patients from the non-HCC individuals were 0.718, 0.682, and 0.642, respectively. The sensitivities were 67.85%, 43.71%, and 44.45%, and the specificities were 75.70%, 92.78%, and 83.91%, respectively. The pairwise comparison of AUCs showed that the diagnostic efficacy of plasma miR7 detection was significantly better than that of AFP or DCP (P＜0.05). Although its specificity was slightly lower than AFP and DCP, the sensitivity was significantly higher. Among patients negative for both AFP and DCP, miR7 maintained an AUC of 0.728 for diagnosing very early-stage HCC, with 67.82% sensitivity and 77.73% specificity. Conclusions Plasma miR7 testing is a potential molecular marker with high sensitivity and specificity for the differential diagnosis of small hepatic nodules. In patients with very early-stage HCC lacking effective molecular markers (negative for both AFP and DCP), miR7 can serve as a novel and effective molecular marker to assist diagnosis.

Objective:To investigate the change trend of hepatectomy for hepatocellular carcinoma and efficacy of fluorescent laparoscopic hepatectomy in a single center from 2014?2023.Methods:The propensity score matching and retrospective cohort study was conducted. The clinicopathological data of 17 232 patients of hepatocellular carcinoma who were admitted to Zhongshan Hospital of Fudan University from January 2014 to December 2023 were collected. There were 14 404 males and 2 828 females, aged 58(range, 11?94)years. Observation indicators: (1) change trend of hepatectomy for hepatocellular carcinoma and application of fluorescent laparoscopy in 2014?2023; (2) propensity score matching between patients undergoing fluorescent and conventional laparoscopic hepatectomy; (3) intraoperative and postoperative conditions of patients in the fluorescent laparoscopy group and the conventional laparoscopy group after propensity score matching. Comparison of measurement data with skewed distribution between groups was conducted using the Mann-Whitney U test. Comparison of count data between groups was conducted using the chi-square test. Comparison of ordinal data between groups was conducted using the nonparametic rank sum test. Propensity score matching was performed using the 1∶3 nearest neighbor matching method, with the caliper value of 0.01. Results:(1) Change trend of hepatectomy for hepatocellular carcinoma and application of fluorescent laparoscopy in 2014-2023. Among 17 232 patients of hepatocellular carcinoma, 4 074 cases underwent laparoscopic surgery, and 13 158 cases underwent open surgery or conversion from laparoscopic to open surgery. From 2014 to 2023, the proportions of laparos-copic surgery in hepatectomy were 8.993%(117/1 301), 6.804%(99/1 455), 9.379%(145/1 546), 17.511%(287/1 639), 23.828%(427/1 792), 24.572%(459/1 868), 29.662%(544/1 834), 31.568%(584/1 850), 38.080%(599/1 573), and 34.246%(813/2 374), respectively. The proportions of Grade Ⅲ surgery in laparoscopic hepatectomy were 11.966%(14/117), 7.071%(7/99), 12.414%(18/145), 21.254%(61/287), 28.337%(121/427), 27.887%(128/459), 34.375%(187/544), 34.644%(214/584), 35.726%(214/599), and 42.681%(347/813), respectively. From 2016 to 2023, the proportions of the application of fluorescent laparoscopy in laparoscopic hepatectomy were 2.069%(3/145), 5.923%(17/287), 8.431%(36/427), 10.022%(46/459), 17.647%(96/544), 24.829%(145/584), 9.182%(55/599), and 12.915%(105/813), respectively. (2) Propensity score matching between patients undergoing fluorescent and conventional laparoscopic hepatectomy. Among the 4 074 patients who underwent laparoscopic liver resection, 503 cases using fluorescent laparoscopy were assigned to the fluorescent laparoscopy group, 3 571 cases using conventional laparoscopy were assigned to the conventional laparoscopy group. A total of 1 794 patients were successfully matched, with 483 cases in the fluorescent laparoscopy group and 1 311 cases in the conventional laparoscopy group. After propensity score matching, the confounding biases of the maximum tumor diameter and the year of surgery were eliminated, and the two groups were comparable. (3) Intraoperative and postoperative conditions of patients in the fluorescent laparoscopy group and the conventional laparoscopy group after propensity score matching. After propensity score matching, there was no perioperative death in either the fluorescent laparoscopy group or the conventional laparoscopy group.The operation time of patients in the fluorescent laparoscopy group was 240(150,328)minutes, the portal triad clamping time was 30(15,45)minutes, the volume of intraoperative blood loss was 50(30,100)mL, the number of patients with complication 72, respectively. For patients in the conventional laparoscopy group, the above indicators were 180(131,240)minutes, 23(15,30)minutes, 55(50,100)mL, and 248, respectively. There were significant differences in the above indicators between the two groups ( Z=?7.593, ?7.372, ?4.941, χ2=3.873, P<0.05). Conclusion:The proportion and difficulty level of laparoscopic hepatectomy for hepatocellular carcinoma in Zhongshan Hospital of Fudan University have shown an upward trend during 2014?2023. Compared with conventional laparoscopy, patients undergoing fluorescent laparoscopic hepatectomy have longer operation time and portal triad clamping time, but less intraoperative blood loss and lower iproportion of complications.

Objective:To investigate the change trend of hepatectomy for hepatocellular carcinoma and efficacy of fluorescent laparoscopic hepatectomy in a single center from 2014?2023.Methods:The propensity score matching and retrospective cohort study was conducted. The clinicopathological data of 17 232 patients of hepatocellular carcinoma who were admitted to Zhongshan Hospital of Fudan University from January 2014 to December 2023 were collected. There were 14 404 males and 2 828 females, aged 58(range, 11?94)years. Observation indicators: (1) change trend of hepatectomy for hepatocellular carcinoma and application of fluorescent laparoscopy in 2014?2023; (2) propensity score matching between patients undergoing fluorescent and conventional laparoscopic hepatectomy; (3) intraoperative and postoperative conditions of patients in the fluorescent laparoscopy group and the conventional laparoscopy group after propensity score matching. Comparison of measurement data with skewed distribution between groups was conducted using the Mann-Whitney U test. Comparison of count data between groups was conducted using the chi-square test. Comparison of ordinal data between groups was conducted using the nonparametic rank sum test. Propensity score matching was performed using the 1∶3 nearest neighbor matching method, with the caliper value of 0.01. Results:(1) Change trend of hepatectomy for hepatocellular carcinoma and application of fluorescent laparoscopy in 2014-2023. Among 17 232 patients of hepatocellular carcinoma, 4 074 cases underwent laparoscopic surgery, and 13 158 cases underwent open surgery or conversion from laparoscopic to open surgery. From 2014 to 2023, the proportions of laparos-copic surgery in hepatectomy were 8.993%(117/1 301), 6.804%(99/1 455), 9.379%(145/1 546), 17.511%(287/1 639), 23.828%(427/1 792), 24.572%(459/1 868), 29.662%(544/1 834), 31.568%(584/1 850), 38.080%(599/1 573), and 34.246%(813/2 374), respectively. The proportions of Grade Ⅲ surgery in laparoscopic hepatectomy were 11.966%(14/117), 7.071%(7/99), 12.414%(18/145), 21.254%(61/287), 28.337%(121/427), 27.887%(128/459), 34.375%(187/544), 34.644%(214/584), 35.726%(214/599), and 42.681%(347/813), respectively. From 2016 to 2023, the proportions of the application of fluorescent laparoscopy in laparoscopic hepatectomy were 2.069%(3/145), 5.923%(17/287), 8.431%(36/427), 10.022%(46/459), 17.647%(96/544), 24.829%(145/584), 9.182%(55/599), and 12.915%(105/813), respectively. (2) Propensity score matching between patients undergoing fluorescent and conventional laparoscopic hepatectomy. Among the 4 074 patients who underwent laparoscopic liver resection, 503 cases using fluorescent laparoscopy were assigned to the fluorescent laparoscopy group, 3 571 cases using conventional laparoscopy were assigned to the conventional laparoscopy group. A total of 1 794 patients were successfully matched, with 483 cases in the fluorescent laparoscopy group and 1 311 cases in the conventional laparoscopy group. After propensity score matching, the confounding biases of the maximum tumor diameter and the year of surgery were eliminated, and the two groups were comparable. (3) Intraoperative and postoperative conditions of patients in the fluorescent laparoscopy group and the conventional laparoscopy group after propensity score matching. After propensity score matching, there was no perioperative death in either the fluorescent laparoscopy group or the conventional laparoscopy group.The operation time of patients in the fluorescent laparoscopy group was 240(150,328)minutes, the portal triad clamping time was 30(15,45)minutes, the volume of intraoperative blood loss was 50(30,100)mL, the number of patients with complication 72, respectively. For patients in the conventional laparoscopy group, the above indicators were 180(131,240)minutes, 23(15,30)minutes, 55(50,100)mL, and 248, respectively. There were significant differences in the above indicators between the two groups ( Z=?7.593, ?7.372, ?4.941, χ2=3.873, P<0.05). Conclusion:The proportion and difficulty level of laparoscopic hepatectomy for hepatocellular carcinoma in Zhongshan Hospital of Fudan University have shown an upward trend during 2014?2023. Compared with conventional laparoscopy, patients undergoing fluorescent laparoscopic hepatectomy have longer operation time and portal triad clamping time, but less intraoperative blood loss and lower iproportion of complications.

ObjectiveTo investigate the effects of different cultivation modes on the yield of Gentiana crassicaulis and its microbial diversity and secondary metabolite content in the rhizosphere soil. MethodWith G. crassicaulis of different cultivation modes and its rhizosphere soil as the research objects， the composition of bacterial and fungal communities， dominant bacteria， and differential microorganisms in the rhizosphere soil were analyzed by high-throughput sequencing technology. HPLC was used to determine the content of iridoids in G. crassicaulis with different cultivation modes. ResultCompared with plastic film mulching， planting without mulch and intercropping of peony， white kidney bean， potato， and corn increased the yield of fresh products by 16.11%-17.68%， 22.48%-26.34%， 29.37%-32.19%， 34.82%-36.57%， and 35.34%-39.71%， respectively， and increased the yield of dry products by 19.75%-23.17%， 25.86%-29.32%， 30.18%-34.94%， 35.22%-39.87%， and 39.72%-43.73%. The total content of four iridoids， including gentiopicrin， loganic acid， sweroside， and swertiamarin， increased by 10.17%-37.83%， 5.93%-47.44%， 9.09%-28.84%， and 10.71%-28.57%， respectively. The diversity of bacterial and fungal communities in the rhizosphere soil increased significantly （P<0.05）. The relative abundance of pathogenic bacteria such as Sordariomycetes， Leotiomycetes， Tremellomycetes， Eurotiomycetes， Fusarium， and Cladophialophora decreased， and the proportions of beneficial bacteria such as Proteobacteria， Acidobacteria， and Actinobacteriota increased and they gradually became the dominant bacteria. ConclusionDifferent cultivation modes can affect the yield of G. crassicaulis and its microbial diversity and iridoid content in the rhizosphere soil. Cultivation without mulch and intercropping patterns have certain advantages， which can provide theoretical references for the planting of G. crassicaulis.

Objective To investigate the effect of exosomes derived from Echinococcus multilocularis on macrophage polarization after treatment for different durations and concentrations. Methods A total of 60 BALB/c mice were used for modeling, among which 4 mice were selected to observe the growth of abdominal lesions on 7.0T MRI. The mice for modeling were dissected, and the protoscoleces was taken from the abdominal lesion and cultured in vitro ; ultracentrifugation was used to extract the exosomes from the supernatant, and transmission electron microscopy and Western blotting were used for the characterization of exosomes. The macrophages without exosome treatment were established as control group, and the macrophages co-cultured with different concentrations of exosomes derived from Echinococcus multilocularis were established as experimental group (10 μg/mL group and 50 μg/mL group) and were cultured for 48 and 72 hours. The morphological changes of macrophages were observed under a microscope, and flow cytometry and ELISA were used to observe polarization state. A one-way analysis of variance was used for comparison of normally distributed continuous data between multiple groups, and the least significant difference t -test was used for further comparison between two groups. Results The results of 7.0T MRI showed the formation of diffuse lesions with different sizes in the abdominal cavity of mice, and the exosomes derived from Echinococcus multilocularis were approximately 100 nm in diameter and were cup-shaped or saucer-shaped, with the positive expression of the surface markers CD9, TSG101, and CD63. After co-culture, most of the cells in the experimental group were elongated with an irregular and polygonal shape. Flow cytometry showed that after 48 hours of co-culture, the positive rates of CD16/32, CD206, and CD369 in the control group were 99.53%±0.06%, 90.27%±0.21%, and 2.40%±0.20%, respectively; compared with the control group, except that the 10 μg/mL exosome group had a significant reduction in the positive rate of CD369 (0.80%±0.00%) ( P < 0.05), all the other groups had a significant increase in the positive rates of CD16/32, CD206, and CD369 (all P < 0.000 1); after 72 hours of co-culture, the positive rates of CD16/32, CD206, and CD369 in the control group were 99.67%±0.06%, 85.47%±0.55%, and 6.60%±0.20%, respectively, and compared with the control group, the experimental group had significant increases in the positive rates of CD16/32, CD206, and CD369 (all P < 0.05). ELISA showed that after 48 hours of co-culture, the levels of IL-6 and TNFα in the control group were 58.53±15.52 pg/mL and 320.70±5.30 pg/mL, respectively, and when the exosome concentration was 50 μg/mL, the level of IL-6 in the experimental group was 98.81±15.55 pg/mL, which was higher than that in the control group ( P < 0.05); after 72 hours of co-culture, the levels of IL-6 and TNFα in the control group were 76.22±9.68 pg/mL and 323.90±87.37 pg/mL, respectively, and when the exosome concentration was 10 μg/mL, the level of TNFα was 164.20±14.17 pg/mL, which was significantly lower than that in the control group ( P < 0.05); when the exosome concentration was 50 μg/mL, the level of IL-6 was 99.52±8.35 pg/mL, which was significantly higher than that in the control group ( P < 0.05). Conclusion Exosomes derived from Echinococcus multilocularis can regulate macrophage polarization and induce M2-like polarization of macrophages after co-culture at a concentration of 10 μg /mL for 72 hours, and further studies are needed to clarify the specific method.

Objective : To investigate the effect of hypoxia/cold exposure on white fat browning in an obese rat model . Methods : Obese rat model was constructed by high⁃fat feeding and randomly divided into control group , of body weight and body fat were observed , and the morphological changes of rat adipose tissue cells were observed using HE staining and compared with the area of adipocytes . The expression of peroxisome proliferator⁃activated receptor γ2 (PPAR⁃γ2) , PR structural domain binding factor 16 ( PRDM16) and uncoupling protein 1 ( UCP⁃1) gene and the expression of UCP⁃1 protein in the adipose tissue of rats were examined by immunofluorescence , realtime fluorescence quantitative PCR and Western blot . Results : Compared with the control group , the body weight and body fat of rats in the hypoxic , hypothermic and cryogenic hypoxic groups were reduced , and the body weight and body fat of rats in the hypothermic hypoxic group were lower than those in the hypoxic and cryogenic groups (P< 0. 05) . Compared with the control group , the adipocyte area of rats in the hypoxia , hypothermia and low⁃temperature hypoxia groups was reduced , and the adipocyte area of rats in the low⁃temperature hypoxia group was lower than that in the hypoxia and low⁃temperature groups (P < 0. 05) . Compared with the control group , the expression of PPAR⁃γ2 , PRDM16 and UCP⁃1 genes all increased in the scapular brown adipose tissue ( F = 378. 495 , 102. 061 , 322. 443 , P < 0. 05) and decreased in the perirenal white adipose tissue (F = 4. 555 , P < 0. 05) in the hypoxic , hypothermic and low⁃temperature hypoxic groups of rats , PRDM16 and UCP⁃1 gene expression all increased (F = 24. 387 , 163 . 660 , P < 0. 05) . Compared with the control group , the expression of UCP⁃1 protein in scapular brown adipose tissue and perirenal white adipose tissue of rats in the hypoxic , hypothermic and low⁃temperature hypoxic groups increased (P < 0. 05) ; UCP⁃1 protein expression was lower in scapular brown adipose tissue and perirenal white adipose tissue of rats in the hypoxic and hypothermic groups compared to the hypothermic hypoxic group (P < 0. 05) . Conclusion Hypoxia/cold exposure can induce white adipose browning and affect the body weight of rats by modulating the intra⁃adipose PPAR⁃γ2 and PRDM16 pathways which lead to high UCP⁃1 expression .