Combining single-cell RNA sequencing and population-based studies reveals hand osteoarthritis-associated chondrocyte subpopulations and pathways

Hand osteoarthritis is a common heterogeneous joint disorder with unclear molecular mechanisms and no disease-modifying drugs.In this study,we performed single-cell RNA sequencing analysis to compare the cellular composition and subpopulationspecific gene expression between cartilage with macroscopically confirmed osteoarthritis(n=5)and cartilage without osteoarthritis(n=5)from the interphalangeal joints of five donors.Of 105142 cells,we identified 13 subpopulations,including a novel subpopulation with inflammation-modulating potential annotated as inflammatory chondrocytes.Fibrocartilage chondrocytes exhibited extensive alteration of gene expression patterns in osteoarthritic cartilage compared with nonosteoarthritic cartilage.Both inflammatory chondrocytes and fibrocartilage chondrocytes showed a trend toward increased numbers in osteoarthritic cartilage.In these two subpopulations from osteoarthritic cartilage,the ferroptosis pathway was enriched,and expression of iron overload-related genes,e.g.,FTH1,was elevated.To verify these findings,we conducted a Mendelian randomization study using UK Biobank and a population-based cross-sectional study using data collected from Xiangya Osteoarthritis Study.Genetic predisposition toward higher expression of FTH1 mRNA significantly increased the risk of hand osteoarthritis(odds ratio=1.07,95%confidence interval:1.02–1.11)among participants(n=332668)in UK Biobank.High levels of serum ferritin(encoded by FTH1),a biomarker of body iron overload,were significantly associated with a high prevalence of hand osteoarthritis among participants(n=1241)of Xiangya Osteoarthritis Study(P-for-trend=0.037).In conclusion,our findings indicate that inflammatory and fibrocartilage chondrocytes are key subpopulations and that ferroptosis may be a key pathway in hand osteoarthritis,providing new insights into the pathophysiology and potential therapeutic targets of hand osteoarthritis.

Hypertonic saline resuscitation maintains a more balanced profile of T-lymphocyte subpopulations in a rat model of hemorrhagic shock

Objective: To investigate the potential and early effect of hypertonic saline resuscitation on T-lymphocyte sub- populations in rats with hemorrhagic shock. Methods: A model of rat with severe hemorrhagic shock was established in 18 Sprague-Dawley (SD) rats. The rats were randomly divided into Sham group, HTS group (hypertonic saline resuscitation group) and NS group (normal saline resuscitation group). Each group contained 6 rats. The CD4+ and CD8+ subpopulations of T-lymphocytes in peripheral blood were detected respectively before shock and after resuscitation by double antibody labelling and flow cytometry. Results: In the early stage after hemorrhagic shock, fluid resuscitation and emergency treatment, the CD4+ lymphocytes of peripheral blood in HTS and NS groups markedly increased. Small volume resuscitation with HTS also induced peripheral CD8+ lymphocytes to a certain extent, whereas NS resuscitation showed no effect in this respect. Consequently, compared with Sham and HTS groups, CD4+/CD8+ ratio of peripheral blood in NS group was obviously increased, and showed statistically differences. Conclusion: In this model of rat with severe hemorrhagic shock, small volume resuscitation with HTS is more effective than NS in reducing immunologic disorders and promoting a more balanced profile of T-lymphocyte subpopula- tions regulating network.

Peripheral T-lymphocyte subpopulations in different clinical stages of chronic HBV infection correlate with HBV load

AIM： To characterize the peripheral T-cell subpopulation profiles and their correlation with hepatitis B virus （HBV） replication in different dinical stages of chronic HBV infection. METHODS： A total of 422 patients with chronic HBV infection were enrolled in this study. The patients were divided into three stages： immune-tolerant stage, immune active stage, and immune-inactive carrier stage. Composition of peripheral T-cell subpopulations was determined by flow cytometry. HBV markers were detected by enzyme-linked immunosorbent assay. Serum HBV DNA load was assessed by quantitative real-time poiymerase chain reaction.RESULTS： CD8^＋ T-cells were significantly higher in patients at the immune-tolerant stage than in patients at the immune-active and -inactive carrier stages （36.87 ± 7.58 vs 34.37 ± 9.07, 36.87 ± 7.58 vs 28.09 ± 5.64, P 〈 0.001）. The peripheral blood in patients at the immune-tolerant and immune active stages contained more CD8^＋ T-cells than CD4^＋ T-cells （36.87 ± 7.58 vs 30.23 ± 6.35, 34.37 ± 9.07 vs 30.92 ± 7.40, P 〈 0.01）, whereas the peripheral blood in patients at the immune- inactive carrier stage and in normal controls contained less CD8^＋ T-cells than CD4^＋ T-cells （28.09 ± 5.64 vs 36.85 ±6.06, 24.02 ± 4.35 vs 38.94 ± 3.39, P 〈 0.01）. ANOVA linear trend test showed that CD8^＋ T-cells were significantly increased in patients with a high viral load （39.41 ± 7.36, 33.83 ± 7.50, 31.81 ± 5.95 and 26.89 ± 5.71, P 〈 0.001）, while CD4^＋ T-cells were significantly increased in patients with a low HBV DNA load （37.45 ± 6.24, 33.33 ± 5.61, 31.58 ± 6.99 and 27.56 ± 5.49, P 〈 0.001）. Nultiple regression analysis displayed that log copies of HBV DNA still maintained its highly significant coefficients for T-cell subpopulations, and was the strongest predictors for variations in CD3^＋, CD4^＋ and CD8^＋ cells and CD4^＋/CD8^＋ ratio after adjustment for age at HBV-infection, maternal HBV-infection status, presence of hepatitis B e antigen and HBV mutation.CONCLUSION： Differences in peripheral T-cell subpopulation profiles can be found in different clinical stages of chronic HBV infection. T-cell impairment is significantly associated with HBV load.

Flow cytometric characterizations of leukocyte subpopulations in the peripheral blood of northern pig-tailed macaques （Macaca leonina）

Pig-tailed macaques(Macaca nemistrina group) have been extensively used as non-human primate animal models for various human diseases in recent years, notably for AIDS research due to their sensitivity to HIV-1. Northern pig-tailed macaques(M. leonina) are distributed in China and other surrounding Southeast Asia countries. Although northern pig-tailed macaques have been bred on a large scale as experimental animals since 2012, the reference value of normal levels of leukocytes is not available. To obtain such information, 62 blood samples from male and female healthy northern pig-tailed macaques at different ages were collected. The normal range of major leukocyte subpopulations, such as T lymphocytes, B lymphocytes, natural killer(NK) cells, monocytes, and the expression levels of activation or differentiation related molecules(CD38, HLA-DR, CCR5, CD21, IgD, CD80 and CD86) on lymphocytes were analyzed by flow cytometry. The counts of B cells decreased with age, but those of CD8+ T cells and NK cells and the frequency of CD38+HLA-DR+CD4+ T cells were positively correlated with age. The counts of leukocyte subpopulations were higher in males than those in females except for CD4+ T cells. Males also showed higher expression levels of Ig D and CD21 within B cells. This study provides basic data about the leukocyte subpopulations of northern pig-tailed macaques and compares this species with commonly used Chinese rhesus macaques(M. mulatta), which is meaningful for the biomedical application of northern pig-tailed macaques.

Hepatitis B virus DNA is more powerful than HBeAg in predicting peripheral T-lymphocyte subpopulations in chronic HBV-infected individuals with normal liver function tests

AIM:To investigate the peripheral T-lymphocyte subpopulation profile,and its correlations with hepatitis B virus(HBV) replication level in chronic HBV-infected(CHI) individuals with normal liver function tests(LFTs) . METHODS:Frequencies of T-lymphocyte subpopu-lations in peripheral blood were measured by flow cytometry in 216 CHI individuals. HBV markers were detected with ELISA. Serum HBV DNA load was assessed with quantitative real-time PCR. Information of age at HBV infection,and maternal HBV infection status was collected. ANOVA linear trend test and linear regression were used in statistical analysis. RESULTS:CHI individuals had significantly decreased relative frequencies of CD3+,CD4+ subpopulationsand CD4+/CD8+ ratio,and increased CD8+ subset percentage compared with uninfected individuals(all P < 0.001) . There was a significant linear relationship between the load of HBV DNA and the parameters of T-lymphocyte subpopulations(ANOVA linear trend test P < 0.01) . The parameters were also significantly worse among individuals whose mothers were known to be HBV carriers,and those having gained infection before the age of 8 years. In multiple regressions,after adjustment for age at HBV infection and status of maternal HBV infection,log copies of HBV DNA maintained its highly significant predictive coefficient on T-lymphocyte subpopulations,whereas the effect of HBeAg was not significant. CONCLUSION:HBV DNA correlates with modification in the relative T-lymphocyte subpopulation frequencies. High viral load is more powerful than HBeAg in predicting the impaired balance of T-cell subsets.

Early Changes of Peripheral Blood Lymphocyte Subpopulations in Patients with Occupational 2,4-dinitrophenol Poisoning

2,4-dinitrophenol （DNP）, an organic compound which frequently used in industry, is considered to have high toxicity. This study aimed to investigate the early changes of lymphocyte subpopulations in patients with occupational 2,4-DNP poisoning. Totally 9 patients with acute occupational 2,4-DNP poisoning and 30 healthy volunteers as control were enrolled. The patients received immediately comprehensive supportive treatments, including large-dose glucocorticoid and repeated hemoperfusion （HP）. The ratio of CD4＋/CD8＋ T cells were significantly higher in patients upon admission compared to healthy controls （P 〈 0.01）; however, counts of total lymphocytes, CD3＋, CD3＋CD4＋, CD3＋CD8＋, B （CD19＋）, and natural killer （NK） cells （CD16＋CD56＋） were significantly reduced （all P 〈 0.001）. The NK cell count was negatively correlated with initial plasma 2,4-DNP concentration （r = -0.750, P = 0.026）. Thus, acute occupational 2,4-DNP poisoning was accompanied by immediate complex immune cell reactions, especially NK cells might play important role in severe 2,4-DNP poisoning.

Olfactory ensheathing cells for spinal cord repair: crucial differences between subpopulations of the glia

OECs for spinal cord repair： Is repairing the iniured spinal cord by olfactory ensheathing cell （OEC） transplantation pos- sible？ A recent human trial in which a paralysed man regained some function after transplantation of partially purified OECs suggests that this therapy may be a successful approach （Ta- bakow et al., 2014）. In another human trial in which olfactory mucosa lamina propria was transplanted, patients recovered some motor and sensory function （Wang et al., 2015）. While these results show promise, it is clear that improvements are needed to provide patients with increased functional output. Strategies to improve the therapeutic use of OECs may include improving the purification of the OECs used for transplantation, using them in combination with growth factors to combat the inhibitory environment and improve anon growth, the use of nerve bridges, advanced physiotherapy and the use of exo- skeleton robotics to reinforce functional connections. Of all these approaches, it is probably is primarily addressed to ensure crucial that the purity of OECs consistency in outcomes.

Analysis of Genetic Diversity of Four Subpopulations of Chimonobambusa rivularis Yi in Sichuan

A study of the genetic diversity within four subpopulations of Chimonobambusa rivularis Yi and adaptations to altitude, habitat and related factors in Qionglai City of Sichuan province in China, analyzed by random amplified polymorphic DNA technique. 22 random primers were selected in the amplification and 375 repetitive loci with 350 polymorphic loci were produced. The total average percentage of amplification loci was 93.3%. The genetic diversity of every subpopulation was medium on （the percentage of amplification loci was 49.33%-66.67%）. The genetic diversity of high altitude samples of Zhengtiantai was lower than that of low altitude Wutonggang＇s. There was no obvious differentiation between two subpopulations of Zhengtiantais. The correlation is very low between altitude and the amplification loci and the genetic distance, Pearson correlation coefficient was 0.431 and 0.488 （P 〈 0.01）. Through investigation, the primary cause of heredity multiplicity drops was that the habitat had been destroyed by tourism development and other human disturbance.

MINIMUM ATTRIBUTE CO-REDUCTION ALGORITHM BASED ON MULTILEVEL EVOLUTIONARY TREE WITH SELF-ADAPTIVE SUBPOPULATIONS

Attribute reduction is an important process in rough set theory.Finding minimum attribute reduction has been proven to help the user-oriented make better knowledge discovery in some cases.In this paper,an efficient minimum attribute reduction algorithm is proposed based on the multilevel evolutionary tree with self-adaptive subpopulations.A model of multilevel evolutionary tree with self-adaptive subpopulations is constructed,and interacting attribute sets are better decomposed into subsets by the self-adaptive mechanism of elitist populations.Moreover it can self-adapt the subpopulation sizes according to the historical performance record so that interacting attribute decision variables are captured into the same grouped subpopulation,which will be extended to better performance in both quality of solution and competitive computation complexity for minimum attribute reduction.The conducted experiments show the proposed algorithm is better on both efficiency and accuracy of minimum attribute reduction than some representative algorithms.Finally the proposed algorithm is applied to magnetic resonance image(MRI)segmentation,and its stronger applicability is further demonstrated by the effective and robust segmentation results.

Correlation of Quantitative Changes in Bone Marrow Lymphocyte Subpopulations with Some Rhabdomyosarcoma Prognosis Factors in Children

Rationale: The known prognosis factors for rhabdomyosarcoma (RMS) in children do not always explain the unsatisfactory outcome of treatment. Changes in the subpopulation composition of Bone Marrow (BM) effector cells during the development of RMS may indicate new directions for the search for prognostic factors and points for the impact of targeted therapy. Purpose: To identify correlations between quantitative changes in the levels of subpopulations of T, B and NK-lymphocytes of BM and known risk factors for RMS in children. Objects: The study included 31 patients. The main group included 16 patients with RMS, average age—6.8 ± 1.0 years, while children 1 - 10 years old—13 (81.3%), over 10 years old—3 (18.8%) people, girls and boys were 8 people each. The embryonic variant of RMS was established in 10 (62.5%) cases, the alveolar variant—in 4 (25%) cases, in two patients (12.5%), the histological variant was not established. In 12 (75%) patients, an unfavorable localization of the RMS (parameningeal, extremities, prostate, bladder) was revealed, in 4 patients (25%), the localization of the tumor was regarded as favorable. Patients with T2b—13 (81.2%) and T2a—2 (12.5%) stages prevailed. Regional and distant metastases were detected in 10 (52.6%) patients. The comparison group included 15 children in whom the presence of malignant neoplasia was excluded, the average age was 8.4 ± 1.5 years, 11 boys (73.7%) and 4 girls (26.3%). Methods: All patients underwent morphological (myelogram counting) and immunological (quantitative analysis of lymphocytic subpopulations) bone marrow studies. Immunophenotyping in all patients was carried out by direct immunofluorescence using a triple fluorescent label. Results: Significant differences in the levels of subpopulations of BM T-lymphocytes were found when comparing the values of the main group, distributed by localization and histological variant, with the data obtained in the control group of patients. For example, the percentage of CD3+ T cells with the co-stimulatory molecule CD28+ was significantly higher in patients with parameningeal RMS (p = 0.010). Conclusion: Each clinical group of patients has its own individual immunological characteristics. The results obtained by us can be considered indicative and regarded as starting points for further study of the peculiarities of the subpopulation composition of BM in patients with RMS.

T Lymphocyte Subpopulations in Normal Pregnancies and Those Complicated by Eclampsia in Kaduna State, Nigeria

Background: Eclampsia, the occurrence of generalized convulsion(s) in association with signs of preeclampsia [PE] (hypertension and proteinuria) in pregnancy has remained a significant public threat in Nigeria, contributing to maternal and perinatal morbidity and mortality. This study was a comparative cross-sectional study conducted in some selected hospitals in Kaduna State, between April 2014 and November 2015. Subjects and Methods: Blood (3 mls) was collected into an ethylenediaminetetraaccetic acid (EDTA) vacutainer tube from third trimester women diagnosed with eclampsia (EC;n = 38) and healthy pregnant controls (PC;n = 38)—age and parity matched and healthy non-pregnant controls (NPC;n = 38)—age matched. T Cell subpopulations and Complete Blood Count levels were measured by Sysmex, Auto blood analyzer and flow cytometry respectively. Participants with smear positive malaria, seropositive for human immunodeficiency virus (HIV), any other clinical infection or refused consent were excluded from this study. Data obtained were analyzed using analysis of variance (ANOVA) and Post Hoc test. A p-value of less than 0.05 was considered to be significant. Result: Overall, results showed a depressed (mean ± Standard deviation (SD): CD3+ T cell (65.6 ± 15.5%;1225.5 ± 401.5 cell/μL), CD4+ T cell (36.1 ± 8.7%;657.1 ± 189.9 cell/μL), and a low CD4/CD8 value in women with EC (1.4 ± 0.5) and PC (1.5 ± 0.3) compared to NPC (1.9 ± 0.6) control (p < 05), while the total white blood cell count, and differential percentage neutrophils count were noted to be elevated among the eclamptic women (9.8 ± 4.9 × 109/L;70.3 ± 12.0%) compared to PC (6.9 ± 3.6 × 109/L;64.6 ± 8.1%) and NPC (5.6 ± 2.0 × 109/L;48.5 ± 10.7%), p < 0.05. Conclusion: Eclampsia was associated with significantly depressed CD3+ and CD4+ T lymphocyte, and increased percentage differential neutrophil counts.

Identification of Sperm Subpopulations in Water Buffalo Ejaculates： Changes in Cryopreservation Stages and Bull Variation

This study aimed to identify and characterize the sperm subpopulations existing in water buffalo semen using a computer assisted sperm analyzer （CASA）, as well as assess the effects of cryopreservation on the sperm subpopulation structure and evaluate bull variability. The semen of eight Bulgarian Murrah bulls was collected by four times in an interval of one week each. The semen was cryopreserved following a standard protocol and sperm kinematics was assessed. Clustering methods were applied to individual sperms, forming two significantly different （P 〈 0.05） subpopulations （P1 and P2）. Subpopulation P1 represents those spermatozoa that moved most rapidly and progressively （46.29%）, and subpopulation P2 includes spermatozoa with relatively low velocity or poorly motile but with high progressiveness （53.41%）. There was a decline on the population of P1 sperms from fresh （52.52%）, pre-freeze （45.73%） to post-thaw （35.17%） stages and significant difference on the sperm kinematics between P1 and P2. A significant decline in the values of distance, velocity and amplitude of lateral head （ALH） parameters were observed at post-thaw stage, while an increase was observed on trajectory and beat cross frequency （BCF） kinematics. Values of sperm kinematics were also significantly different （P 〈 0.05） among all bulls. The frequency distribution of spermatozoa on both subpopulations P1 and P2 was quite similar for all bulls in pre-freeze and post-thaw stages, but with significant （P 〈 0.05） variability on fresh stage. Bulls with the highest maintained frequency of P1 sperms are denoted as good freezer bulls. In sum, kinematic characterization of water buffalo sperm and clustering into subpopulation enable to identify bulls that are more resistant to cryopreservation and production of quality semen for genetic propagation.

Analysis of Peripheral T-Lymphocyte Subpopulations in Patients with Early Syphilis by Flow Cytometry

Objective: To study the cellular immunity function of patientswith early syphilis and the effects on immune modifiersEsberitox N or IFN. Methods: T-lymphocyte subpopulations of the peripheralblood in 44 patients with syphilis and 40 healthy controls wereexamined by flow cytometry. Results: The number of CD_4+ cells and the CD_4+/CD_8+ ratioin patients with syphilis were found to be significantly lowerthan those in the control (P<0.0l), while the number of CD_8+cells was higher than that in the control (P<0.01). TheCD_4+/CD_8+ ratio in those with active disease was lower thanthat in those who had been cured (P<0.05). The CD_4+ countand the CD_4+/CD_8+ ratio in those treated with antibiotics alone(Penicillin G or Cephalosporins) were lower than those treatedwith both antibiotics and immunomodulators (P<0.05). Conclusions: Cellular immunity in the patients with earlysyphilis was prominently suppressed, and treatment withimmunomodulators may be helpful for the recovery of cellularimmunity of these patients.

流式细胞术检测细胞周期影响因素及不同免疫细胞亚群周期分析

细胞周期检测对于了解细胞增殖状态、细胞功能研究、药物筛选与评估等领域具有重要意义。流式细胞仪分析细胞周期是目前较为经典且广泛应用的检测手段之一。碘化丙啶(propidium iodide,PI)单染是最常用的基于流式的检测细胞周期方法,然而,该方法在使用过程有诸多处理因素会对检测结果造成影响。此外,不同免疫细胞亚群在不同细胞周期阶段的分布差异也有助于研究免疫反应,了解疾病状态。本文以B16-F10细胞系为例,采用PI单染法,从固定条件、上机条件和软件分析3个角度,评价多种处理因素对细胞周期检测结果的影响。根据结果分析,在进行细胞周期检测时,建议取3×10^(6)个细胞,用300μL预冷PBS重悬细胞后逐滴滴入700μL预冷的无水乙醇,放置于4℃或-20℃过夜固定,低速收样,收样速率为每秒400~600颗粒数,去黏连后需收集至少3000个细胞。另外,通过EdU、PI双指标染色可以精确划分细胞周期,而细胞表面标志物染色联合Ki-67和PI染色法,可在不进行细胞分选的情况下进行不同免疫细胞亚群周期分析。本文较为系统的探讨了PI单染法检测细胞周期的影响因素,提供了标准的实验操作流程。建立了EdU联合PI检测细胞周期和针对不同免疫细胞亚群周期分析方案,拓宽了细胞周期检测方式。

鼻咽癌放化疗治疗患者外周血PD-1及免疫指标水平的变化及其临床意义

目的探讨鼻咽癌放化疗治疗患者外周血程序性死亡受体1(PD-1)及免疫指标水平的变化及其临床意义。方法回顾性选取2020年1月至2023年江苏省肿瘤医院收治的90例鼻咽癌患者,所有患者均经病理确诊并接受放化疗治疗,采集其血液样本之后采用流式细胞术对不同时间段(治疗前、新辅助化疗后、放疗后)外周血淋巴细胞亚群比例、外周血PD-1、CD8^(+)CD28^(+)细胞比例的水平变化予以动态监测并比较。结果鼻咽癌患者新辅助化疗后CD3^(+)、CD4^(+)细胞比例、CD4^(+)/CD8^(+)比值分别为(72.28±8.37)%、(39.27±8.58)%、1.58±0.67,均明显高于治疗前,CD3-CD16^(+)CD56^(+)、CD19^(+)细胞比例分别为(18.27±8.38)%、(7.87±4.08)%,均明显低于治疗前,差异均有统计学意义(P<0.05);新辅助化疗后与治疗前的CD8^(+)细胞比例比较,差异无统计学意义(P>0.05)。鼻咽癌患者新辅助化疗之后CD8^(+)CD28^(+)细胞比例为(10.68±3.87)%,明显高于治疗前,差异有统计学意义(P<0.05);新辅助化疗后与治疗前的外周血PD-1水平比较,差异无统计学意义(P>0.05)。放疗后与治疗前的CD3^(+)细胞比例比较,差异无统计学意义(P>0.05);鼻咽癌患者放疗后CD4^(+)细胞比例、CD4^(+)/CD8^(+)比值、CD19^(+)细胞比例分别为(26.68±6.09)%、0.88±0.29、(3.69±2.36)%,均明显低于治疗前,CD8^(+)、CD3-CD16^(+)CD56^(+)细胞比例分别为(31.03±8.08)%、(27.39±10.26)%,均明显高于治疗前,差异均有统计学意义(P<0.05)。鼻咽癌患者放疗后CD8^(+)CD28^(+)细胞比例为(7.08±2.57)%,明显低于治疗前,外周血PD-1水平为(13.38±6.27)%,明显高于治疗前,差异均有统计学意义(P<0.05)。结论新辅助化疗之后鼻咽癌患者外周血T细胞亚群比值处于持续上调趋势,而放疗完成后处于下降趋势,表明鼻咽癌患者于放疗完成后免疫功能受损;放疗完成后鼻咽癌患者T细胞PD-1表达水平明显上调,提示PD-1抑制剂最佳使用时间可能为放化疗完成时,抗PD-1维持治疗可发挥持久、高效的抗肿瘤作用。

腹腔镜胰十二指肠切除术治疗早期胰头癌患者的效果

目的:观察腹腔镜胰十二指肠切除术(LPD)治疗早期胰头癌患者的效果。方法:选取2022年8月至2023年10月该院收治的92例早期胰头癌患者进行前瞻性研究,按照随机数字表法将其分为对照组与观察组各46例。对照组采用开腹胰十二指肠切除术(OPD)治疗,观察组采用LPD治疗。比较两组围术期相关指标(手术时间、术中出血量、淋巴结清扫数目、术后排气时间、下床时间、住院时间及术后疼痛程度)水平、手术前后T细胞亚群指标(CD4+、CD8+、CD4+/CD8+)水平、R0切除率、半年复发率及并发症发生率。结果:两组淋巴结清扫数目比较,差异无统计学意义(P>0.05);观察组术中出血量少于对照组,手术时间长于对照组,术后排气时间、下床时间及住院时间短于对照组,视觉模拟评分法评分低于对照组,差异均有统计学意义(P<0.05);术后,两组CD4+、CD4+/CD8+水平均低于术前,但观察组高于对照组,两组CD8+水平均高于术前,但观察组低于对照组,差异有统计学意义(P<0.05);两组R0切除率、半年复发率、并发症发生率比较,差异均无统计学意义(P>0.05)。结论:与传统OPD相比,LPD治疗早期胰头癌患者可减少术中出血量,缩短术后排气时间、下床时间及住院时间,减轻术后疼痛程度,且术中淋巴结清扫数目、R0切除率、半年复发率、并发症发生率与OPD相当,但会延长手术时间。

金匮肾气丸联用cART对HIV-1感染者的疗效及不良反应分析

获得性免疫缺陷综合征(acquired immunodeficiency syndrome,AIDS)的主要治疗方法为联合抗逆转录病毒治疗(combined antiretroviral therapy,cART),但部分患者由于免疫功能重建不全和不良反应,可能出现药物性肝损伤、消化功能下降、神经系统毒性和代谢紊乱等,严重影响了抗病毒疗效和患者生存质量,已成为AIDS治疗领域的瓶颈[1]。部分国家资助的中医药防治AIDS科研课题提出正虚(主要为肾虚)是AIDS发生发展的内在病理基础,以肾气与肾阴肾阳的功能下降为主要特征[2-3]。补肾制剂可能是缓解人类免疫缺陷病毒(human immunodeficiency virus,HIV)感染患者病情和不良反应的可靠方式。金匮肾气丸是由地黄、山药、茯苓、牡丹皮、山茱萸、牛膝、泽泻等多味中药制成的综合制剂[4],具有温补肾阳、化气行水的作用。本研究首次采用金匮肾气丸与cART联用治疗HIV-1感染者,比较了金匮肾气丸与cART联用、单用cART对HIV-1感染者抗病毒疗效和不良反应的差异,试图为探索HIV-1感染者治疗的新途径提供参考。

人脐带间充质干细胞的异质性探讨及基于单细胞RNA测序结果的亚群分析

目的探讨人脐带间充质干细胞(hUCMSCs)的异质性,并基于单细胞RNA测序结果进行亚群分析。方法选择两株由郑州奥博细胞医学实验室有限公司提供的hUCMSCs(分别命名为hUCMSCs-1和hUCMSCs-2),通过实时荧光定量聚合酶链反应(qPCR)进行干性基因[性别决定区Y-box 2(Sox2)、Nanog]检测。对hUCMSCs进行成骨诱导,3 d后检测成骨基因碱性磷酸酶(Alp)的表达,7 d后进行碱性磷酸酶染色,14 d后进行茜素红染色。对两株hUCMSCs进行成软骨诱导,21 d后检测成软骨基因二型胶原(COL2A1)的表达。用10 ng/mL白细胞介素-1β(IL-1β)刺激大鼠软骨细胞24 h,与hUCMSCs-1和hUCMSCs-2共培养后,检测软骨细胞合成代谢基因蛋白聚糖(ACAN)和分解代谢基因基质金属蛋白酶3(MMP3)的表达。对hUCMSCs-1进行单细胞RNA测序,并根据功能进行亚群分组,通过生物信息学分析方法描绘各个亚群标志基因热图、富集的信号通路。结果两株hUCMSCs的Sox2、Nanog、Alp、COL2A1表达水平差异有统计学意义(P<0.05)。在成骨诱导7 d后,hUCMSCs-1的碱性磷酸酶染色程度深于hUCMSCs-2。在成骨诱导14 d后,茜素红染色结果显示,hUCMSCs-1钙结节数量多于hUCMSCs-2。经IL-1β干预后,软骨细胞与两株hUCMSCs共培养后的MMP3表达水平差异有统计学意义(P<0.05),但ACAN表达水平差异不显著(P>0.05)。基于单细胞RNA测序结果,hUCMSCs-1可分为C1、C2、C3三个功能亚群。C1亚群的信号通路在细胞外基质受体相互作用上富集,与软骨细胞合成代谢相关;C2亚群信号通路在细胞衰老和凋亡上富集,与细胞的自我更新相关;C3亚群信号通路在DNA复制和减数分裂上富集,与细胞周期相关。结论不同个体来源的hUCMSCs存在异质性,其在成骨、成软骨和抗分解代谢能力方面可能存在差异。通过单细胞RNA测序可较好地根据hUCMSCs的功能特性对其进行分群,有助于提高间充质干细胞治疗的临床疗效。

雷火炙结合大活络胶囊对神经根型颈椎病患者炎症因子、血液流变学指标和T淋巴细胞亚群的影响

目的探究雷火灸结合大活络胶囊对神经根型颈椎病(CSR)患者炎症因子、血液流变学指标和T淋巴细胞亚群的影响。方法前瞻性选取2021年10月至2023年10月广州市中西医结合医院收治的120例CSR患者作为研究对象,按照随机数字表法将其分为对照组和观察组,每组各60例。对照组给予大活络胶囊(口服,3次/d,每次服用4粒)治疗,观察组给予雷火灸结合大活络胶囊治疗,雷火灸每次30 min,1次/d,每周5次,两组均持续治疗2周。比较两组患者的临床疗效,治疗前和治疗2周的疼痛程度[视觉模拟评分法(VAS)评分]、症状积分、炎症因子[白细胞介素(IL)-6、IL-8、肿瘤坏死因子-α(TNF-α)]、血液流变学指标[全血高切黏度、全血低切黏度、血浆黏度、红细胞比容(HCT)]及T淋巴细胞亚群(CD3^(+)、CD4^(+)、CD8^(+)、CD4^(+)/CD8^(+))变化。结果观察组临床总有效率为91.67%,高于对照组(76.67%),差异有统计学意义(P<0.05)。治疗2周后,两组VAS均较治疗前降低,症状积分均较治疗前升高,且观察组的VAS评分为(1.74±0.55)分,低于对照组[(2.82±0.83)分],症状积分为(16.20±3.26)分,高于对照组[(14.85±3.47)分],差异均有统计学意义(P<0.05)。治疗2周后,两组血清IL-6、IL-8、TNF-α水平均较治疗前降低,且观察组的血清IL-6、IL-8、TNF-α水平分别为(27.37±3.47)ng/L、(56.54±15.21)g/L、(8.61±2.13)ng/L,均低于对照组[(38.34±4.64)ng/L、(81.91±16.78)g/L、(13.57±2.42)ng/L],差异均有统计学意义(P<0.05)。治疗2周后,两组全血高切黏度、全血低切黏度、血浆黏度及HCT水平均较治疗前降低,且观察组的全血高切黏度、全血低切黏度、血浆黏度及HCT分别为(3.11±0.45)mPa/s、(6.62±2.17)mPa/s、(1.35±0.1)mPa/s、(38.95±3.08)%,均低于对照组[(3.62±0.68)mPa/s、(8.35±2.36)mPa/s、(1.78±0.26)mPa/s、(42.97±2.85)%],差异均有统计学意义(P<0.05)。治疗2周后,CD3^(+)、CD4^(+)水平及CD4^(+)/CD8^(+)均较治疗前升高,CD8^(+)水平均较治疗前降低,且观察组的CD3^(+)、CD4^(+)水平及CD4^(+)/CD8^(+)分别为(75.32±7.23)%、(47.58±6.33)%、2.17±0.77,均高于对照组[(68.04±8.01)%、(42.49±7.47)%、1.79±0.72],CD8^(+)水平为(20.15±4.45)%,低于对照组[(23.75±4.54)%],差异均有统计学意义(P<0.05)。结论雷火灸结合大活络胶囊治疗CSR疗效显著,可有效减轻患者疼痛,缓解临床症状,抑制炎症反应、改善血液流变学指标、增强免疫力可能是其疗效的作用机制。

抑癌止痛散联合曲马多缓释片对原发性肝癌癌性疼痛疗效、肝癌标志物及T细胞亚群的影响

目的:探讨抑癌止痛散联合盐酸曲马多缓释片对原发性肝癌轻中度癌性疼痛患者的治疗效果及对血清肝癌标志物、T细胞亚群的影响。方法:选取我院2021年11月至2022年12月的原发性肝癌轻中度癌性疼痛住院患者60例,按完全随机数字法分为对照组和观察组各30例。两组患者均给予常规治疗,对照组在常规治疗基础上给予盐酸曲马多缓释片治疗,观察组在常规治疗基础上给予抑癌止痛散外敷联合盐酸曲马多缓释片治疗,共治疗10 d。观察两组患者治疗前后疼痛缓解情况(NRS)评分、生活质量(QOL-LC)评分、血清甲胎蛋白(AFP)、甲胎蛋白异质体(AFP-L3)、T淋巴细胞亚群(CD4^(+)、CD8^(+))水平及治疗结束时两组患者盐酸曲马多缓释片平均剂量,并判定临床疗效。结果:治疗10 d后观察组患者临床疼痛总缓解率96.67%,明显高于对照组的73.33%(P<0.05);治疗后两组患者QOL-LC评分均高于治疗前(P<0.05),且观察组优于对照组(P<0.05);治疗后两组患者血清AFP、AFP-L3水平较治疗前下降(P<0.05),且观察组下降幅度更大(P<0.05);治疗后两组患者血清中CD4^(+)含量均较治疗前上升(P<0.05),且观察组上升更显著(P<0.05);治疗后两组患者血清中CD8^(+)含量均较治疗前下降(P<0.05),且观察组下降更显著(P<0.05);观察组患者治疗结束时盐酸曲马多缓释片平均剂量较对照组明显减少,差异具有统计学意义(P<0.05)。结论:抑癌止痛散外敷联合盐酸曲马多缓释片对于原发性肝癌合并轻中度癌性疼痛患者,能改善临床症状,降低肿瘤标志物水平,改善患者免疫功能,减少止痛药物用量,减轻药物副作用,提高患者生活质量,具有较好的临床疗效,值得推广应用。