Background/Aims: Liver cirrhosis involves chronic inflammation and progressive fibrosis.Among various immune cells, CD8+ T cells are considered a major contributor to hepatic inflammation and fibrosis. However, the exact molecular pathways governing CD8+ T-cell-mediated effects in cirrhosis remain unclear. Methods: This study analyzed transcriptomic and single-cell sequencing data to elucidate CD8+ T-cell heterogeneity and implications in cirrhosis. Results: Weighted gene co-expression analysis of bulk RNA-seq data revealed an association between cirrhosis severity and activated T-cell markers like HLA and chemokine genes. Furthermore, single-cell profiling uncovered eight CD8+ T-cell subtypes, notably, effector memory (Tem) and exhausted (Tex) T cells. Tex cells, defined by PDCD1, LAG3, and CXCL13 expression, were increased in cirrhosis, while Tem cells were decreased. Lineage tracing and differential analysis highlighted CXCL13+ Tex cells as a terminal, exhausted subtype of cells with roles in PD-1 signaling, glycolysis, and T-cell regulation. CXCL13+ Tex cells displayed T-cell exhaustion markers like PDCD1, HAVCR2, TIGIT, and TNFRSF9. Functional analysis implicated potential roles of these cells in immunosuppression. Finally, a CXCL13+ Tex-cell gene signature was found that correlated with cirrhosis severity and poorer prognosis of liver cancer. Conclusions In summary, this comprehensive study defines specialized CD8+ T-cell subpopulations in cirrhosis, with CXCL13+ Tex cells displaying an exhausted phenotype associated with immune dysregulation and advanced disease. Key genes and pathways regulating these cells present potential therapeutic targets.

Background/Aims: Liver cirrhosis involves chronic inflammation and progressive fibrosis.Among various immune cells, CD8+ T cells are considered a major contributor to hepatic inflammation and fibrosis. However, the exact molecular pathways governing CD8+ T-cell-mediated effects in cirrhosis remain unclear. Methods: This study analyzed transcriptomic and single-cell sequencing data to elucidate CD8+ T-cell heterogeneity and implications in cirrhosis. Results: Weighted gene co-expression analysis of bulk RNA-seq data revealed an association between cirrhosis severity and activated T-cell markers like HLA and chemokine genes. Furthermore, single-cell profiling uncovered eight CD8+ T-cell subtypes, notably, effector memory (Tem) and exhausted (Tex) T cells. Tex cells, defined by PDCD1, LAG3, and CXCL13 expression, were increased in cirrhosis, while Tem cells were decreased. Lineage tracing and differential analysis highlighted CXCL13+ Tex cells as a terminal, exhausted subtype of cells with roles in PD-1 signaling, glycolysis, and T-cell regulation. CXCL13+ Tex cells displayed T-cell exhaustion markers like PDCD1, HAVCR2, TIGIT, and TNFRSF9. Functional analysis implicated potential roles of these cells in immunosuppression. Finally, a CXCL13+ Tex-cell gene signature was found that correlated with cirrhosis severity and poorer prognosis of liver cancer. Conclusions In summary, this comprehensive study defines specialized CD8+ T-cell subpopulations in cirrhosis, with CXCL13+ Tex cells displaying an exhausted phenotype associated with immune dysregulation and advanced disease. Key genes and pathways regulating these cells present potential therapeutic targets.

Background/Aims: Liver cirrhosis involves chronic inflammation and progressive fibrosis.Among various immune cells, CD8+ T cells are considered a major contributor to hepatic inflammation and fibrosis. However, the exact molecular pathways governing CD8+ T-cell-mediated effects in cirrhosis remain unclear. Methods: This study analyzed transcriptomic and single-cell sequencing data to elucidate CD8+ T-cell heterogeneity and implications in cirrhosis. Results: Weighted gene co-expression analysis of bulk RNA-seq data revealed an association between cirrhosis severity and activated T-cell markers like HLA and chemokine genes. Furthermore, single-cell profiling uncovered eight CD8+ T-cell subtypes, notably, effector memory (Tem) and exhausted (Tex) T cells. Tex cells, defined by PDCD1, LAG3, and CXCL13 expression, were increased in cirrhosis, while Tem cells were decreased. Lineage tracing and differential analysis highlighted CXCL13+ Tex cells as a terminal, exhausted subtype of cells with roles in PD-1 signaling, glycolysis, and T-cell regulation. CXCL13+ Tex cells displayed T-cell exhaustion markers like PDCD1, HAVCR2, TIGIT, and TNFRSF9. Functional analysis implicated potential roles of these cells in immunosuppression. Finally, a CXCL13+ Tex-cell gene signature was found that correlated with cirrhosis severity and poorer prognosis of liver cancer. Conclusions In summary, this comprehensive study defines specialized CD8+ T-cell subpopulations in cirrhosis, with CXCL13+ Tex cells displaying an exhausted phenotype associated with immune dysregulation and advanced disease. Key genes and pathways regulating these cells present potential therapeutic targets.

Background/Aims: Liver cirrhosis involves chronic inflammation and progressive fibrosis.Among various immune cells, CD8+ T cells are considered a major contributor to hepatic inflammation and fibrosis. However, the exact molecular pathways governing CD8+ T-cell-mediated effects in cirrhosis remain unclear. Methods: This study analyzed transcriptomic and single-cell sequencing data to elucidate CD8+ T-cell heterogeneity and implications in cirrhosis. Results: Weighted gene co-expression analysis of bulk RNA-seq data revealed an association between cirrhosis severity and activated T-cell markers like HLA and chemokine genes. Furthermore, single-cell profiling uncovered eight CD8+ T-cell subtypes, notably, effector memory (Tem) and exhausted (Tex) T cells. Tex cells, defined by PDCD1, LAG3, and CXCL13 expression, were increased in cirrhosis, while Tem cells were decreased. Lineage tracing and differential analysis highlighted CXCL13+ Tex cells as a terminal, exhausted subtype of cells with roles in PD-1 signaling, glycolysis, and T-cell regulation. CXCL13+ Tex cells displayed T-cell exhaustion markers like PDCD1, HAVCR2, TIGIT, and TNFRSF9. Functional analysis implicated potential roles of these cells in immunosuppression. Finally, a CXCL13+ Tex-cell gene signature was found that correlated with cirrhosis severity and poorer prognosis of liver cancer. Conclusions In summary, this comprehensive study defines specialized CD8+ T-cell subpopulations in cirrhosis, with CXCL13+ Tex cells displaying an exhausted phenotype associated with immune dysregulation and advanced disease. Key genes and pathways regulating these cells present potential therapeutic targets.

Objective:To investigate the correlations between expression of long noncoding RNA metastasis-associated lung adenocarcinoma transcript 1 (MALAT1), nuclear-enriched abundant transcript 1 (NEAT1) and their functions on exosome secretion, proliferation and invasion in hepatocellular carcinoma (HCC).Methods:We used small interfering RNA of MALAT1 (si-MALAT1) to knockdown MALAT1 in HuH-7. At the meanwhile, cells which were transfected with si-NC were used as the negative control group. Expression of NEAT1, cell proliferation and invasion function were detected these two groups. HuH-7 cells were transfected with lentivirus NEAT1 over expressing vector (lv-NEAT1) or negative control (lv-control). Expression of exosomes secretion related genes were analyzed between lv-NEAT1 and lv-control groups. Cells of lv-NEAT1 were knockdown MALAT1 expression using si-MALAT1, which could be si-MALAT1+ lv-NEAT1 group. exosomes secretion was detected in si-NC, si-MALAT1 and si-MALAT1+ lv-NEAT1 group. We treated cells (si-MALAT1 group) with exosomes from cells with lv-NEAT1 or lv-control to divide cells as si-MALAT1+ exosomes of lv-NEAT1 cells and si-MALAT1+ exosomes of lv-control groups. Cell proliferation and invasion of cells were detected in two groups.Results:Low expression of NEAT1 were found in MALAT1 knockdown cells compared with si-NC group [(0.72±0.02) vs. (0.98±0.01), P<0.05]. Cells with MALAT1 knockdown shown diminished proliferation [(0.66±0.03) vs. (0.98±0.04), P<0.05)] and invasion [(88.33±7.26) vs. (147.70±13.62), P<0.05)]. Compared with si-NC group, CD9 and CD63 expression were decreased in exosomes of si-MALAT1 group. Compared with si-MALAT1 group, CD9 and CD63 expression was increased in exosomes of si-MALAT1+ lv-NEAT1 group. Compared with si-MALAT1+ exosomes of lv-control group, proliferation [(0.97±0.03) vs. (0.74±0.05), P<0.05)] and invasion [ (132.70±7.36) vs. (98.33±6.01), P<0.05) ] were increased in si-MALAT1+ exosomes of lv-NEAT1 group. Exosomes related genes expression including HSPA8 (5.53±0.31), SLC3A2 (0.32±0.07) and SLC7A5 (0.77±0.45) were changed in lv-NEAT1 group compared with lv-control group [(0.98±0.15), P<0.05]. Conclusion:MALAT1 induced exosomes secretion by NEAT1 and exosomes related genes regulation. This regulation might be related with increased proliferation and invasion function in HCC cells with MALAT1 and NEAT1 abnormal expression.

OBJECTIVE： To prepare Syringopicroside solid lipid nanoparticles （SYR-SLN）， and optimize the formula and characterize SYR-SLN. METHODS： SYR-SLN were prepared by emulsion evaporation method. Using entrapment efficiency as index， based on single factor， orthogonal design was adopted to optimize the mass ratio of lecithin-monoglyceride， volume ratio of organ phase to water phase， poloxamer 188 （F68） concentration and drug dosage. The optimal formula technology was established to investigate entrapment efficiency， drug-loading amount， morphology， particle size， Zeta potential， stability， etc. RESULTS： The mass ratio of lecithin-monoglyceride was 3 ∶ 1； the volume ratio of organic phase to water phase was 1 ∶ 2； the concentration of F68 was 0.4％； drug dosage was 10 mg. The optimal formula included that monoglyceride 80 mg， lecithin 240 mg， 0.4％ F68， syringopicroside 10 mg， absolute ethyl alcohol 5 mL， distilled water 10 mL， emulsification temperature at 65℃ and stirring at 600 r/min. Encapsulation efficiency of SYR-SLN was （42.35±0.60）％ （n＝3）； drug-loading amount was （5.33±0.03）％ （n＝3）； SYR-SLN had a spherical morphology and was evenly distributed. The average particle size was （180.30±5.31） nm with Zeta potential of （－41.9±0.8） mV， and the SYR-SLN could maintain stable for 15 days at 4℃. CONCLUSIONS： SYR-SLN is prepared successfully， and the technology is simple with high encapsulation efficiency.

A new caffeate compound, (E)-erythro-syringylglyceryl caffeate (1), was isolated from the roots and rhizomes of Nardostachys chinensis Batal., together with nine known phenolic compounds, including (+)-licarin A (2), naringenin 4', 7-dimethyl ether (3), pinoresinol-4-O-β-D-glucoside (4), caraphenol A (5), Z-miyabenol C (6), protocatechuic acid (7), caffeic acid (8), gallic acid (9) and vanillic acid (10). Their chemical structures were elucidated on the basis of spectroscopic data and physicochemical properties. Furthermore, this is the first report of compounds 2, 5 and 6 from Nardostachys genus.

Objective To investigate the significance of serum neuron specific enolase (NSE) in the severity and prognosis assessments of diffuse axonal injury (DAI).Methods The levels of serum NSE were measured by radioimmunoassay (RIA) at 12 hours and 1st,2nd,3rd,7th and 14th days after injury in 79patients with DAI.The relationship of serum NSE level with the severity and the prognosis of DAI were analyzed in the patients with DAI.Another 15 patients with only limb fracture and without hemorrhagic shock treated in the hospital during the same period served as the control group.Results The serum NSE levels of the mild injury group were (10.47 ± 2.75) ng/L,(13.41 ± 3.45) ng/L,(16.41 ±4.14) ng/L,(15.57 ±4.28) ng/L,(7.95 ±2.79) ng/L,and (6.39 ± 1.55)ng/L at 12 hours and 1st,2nd,3rd,7th and 14th days after injury.The serum NSE levels of the moderate injury group were (14.98 ± 3.78) ng/L,(19.88 ± 4.78)ng/L,(22.41 ±5.50) ng/L,(20.11 ±6.60) ng/L,(14.59 ±6.64) ng/L,and (8.31 ±3.83) ng/L respectively at 12 hours and 1st,2nd,3rd,7th and 14th days after injury.While the serum NSE levels of the severe injury group were (27.22 ± 4.54) ng/L,(36.43 ± 10.38) ng/L,(41.32 ± 12.44) ng/L,(43.98 ±9.51) ng/L,(42.22 ± 13.05) ng/L,and (37.59 ± 12.96) ng/L at 12 hours and 1st,2nd,3rd,7th and 14th days after injury respectively.The NSE levels in each time point were significantly higher in the severe injury group than in the mild and moderate injury groups (F within =28.11,P ＜ 0.001 ; F between =57.34,P ＜0.001 ;F interaction =8.21,P ＜ 0.001 ;P ＜ 0.01).Compared with the control group ((6.26 ± 1.35) ng/L),the serum NSE levels of the DAI group were significantly different at 12 hours after injury ((18.16 ± 3.76)ng/L,t =2.938,P ＜ 0.01).At three months after injury,patients were divided into the decreased group (n =9),poor prognoses group (in vegetative state or severely disabled,n =29) and good prognoses group (moderately disabled or completely recovered,n =41) according to the GOS score.The serum NSE levels of the decreased group were (32.07 ± 5.73) ng/L,(43.12 ± 15.04) ng/L,(48.26 ± 14.89) ng/L,(50.47 ±11.05) ng/L,(52.90 ±3.82) ng/L,and (56.17 ± 14.62) ng/L respectively at 12 hours and 1st,2nd,3rd,7th and 14th days after injury.The serum NSE levels of the poor prognoses group were (21.90 ± 4.95) ng/L,(24.13 ± 9.94) ng/L,(26.43 ± 6.99) ng/L,(21.62 ± 9.77) ng/L,(15.80 ± 7.15) ng/L,and (10.16 ± 2.33) ng/L respectively at 12 hours and 1st,2nd,3rd,7th and 14th days after injury.The serum NSE levels of the good prognoses group were (13.61 ±4.56) ng/L,(13.75 ±5.10) ng/L,(14.77 ±5.41) ng/L,(13.47 ±4.49) ng/L,(8.92 ± 5.61) ng/L,and (6.60 ± 2.30) ng/L at 12 hours and 1 st,2nd,3rd,7th and 14th days after injury respectively.At each time point,the serum NSE levels were significantly different in the decreased group than in the good prognoses and the poor prognoses groups (F within =18.70,P ＜ 0.001 ; F between =62.97,P ＜0.001 ;F interaction =11.83,P ＜0.001).Conclusion The serum NSE levels can be regard as an index for judging the injury severity and prognosis of DAI,and can be used to guide the option and adjustment of therapeutic approaches for patients with DAI.

Objective To compare the prophylactic effect of laminar closure between titanium miniplate and anchor fixation in open-door cervical laminoplasty.Methods Between January 2010 and December 2010,63 patients with cervical spondylotic myelopathy were treated by open-door laminoplasty.Of them,30 patients underwent laminoplasty by titanium miniplate fixation and 33 by anchor fixation.During follow-up,multi-detector CT was performed preoperatively,at 1 week and 6 months after surgery.At each level,the anteroposterior diameter (APD) of the spinal canal and opening angle (OA) were measured.And the spinal canal expansion rate are calculated.MRI was performed preoperatively and 1 year after surgery to evaluate the severity of cord compression.Results All incisions healed by first intention.The incidence of postoperative axial symptoms in miniplate fixation group and anchor fixation group were 33.3％ (10/30) and 39.4％ (13/33),respectively.The OA,APD,and the spinal canal expansion rate of patients in both groups improved significant postoperatively,but differing from miniplate fixation group.The OA,the APD and the spinal canal expansion rate in anchor fixation group after 6 months were reduced than one week after surgery,and the difference between the groups was statistically significant.Lamina close in two groups was not found.CT images at 6 months showed complete fusion of the hinge area by mature bone or callus in two groups,by cervical sagittal MRI assessment.The severity of spinal cord compression was improved after 1 year.Preoperative and 1 year after the surgery,the severity of spinal cord compression between the two groups showed no significant difference.The severity of spinal cord compression after 1 year in both groups were no more than three grade.Conclusion Open-door cervical laminoplasty by anchor fixation or titanium miniplate can effectively prevent the occurrence of postoperative lamina closure,which can help patients to do functional exercises early,but improvement of spinal cord compression has no significant difference between both of them.However,titanium miniplate fixation for maintenance of the expansive spinal canal is better.