This study investigates the expression pattern and functional significance of EF-hand domain-containing protein 2 (EFHD2) in hepatocellular carcinoma (HCC), with particular focus on its regulatory effects on tumor proliferation, migration, and invasion. Cellular experimental study was completed from June 2024 to January 2025 in the Basic Laboratory of the General Hospital of Southern Theater Command. TCGA database to determine EFHD2 expression and its clinicopathological correlations. GSCA database to assess methylation patterns and immune infiltration. Model of transient overexpression and knockdown of EFHD2 was constructed in hepatocellular carcinoma cells Hep3B, then RT-qPCR and Western blot were applied to verify the transfection efficiency. CCK-8 and colony formation assays for proliferation assessment, Transwell chambers for migration/invasion quantification. Protein-protein interaction networks were constructed via STRING, followed by GO/KEGG enrichment analysis. Statistical analysis was performed using the two independent samples t-test. The results showed that EFHD2 demonstrated significant upregulation in HCC tissues versus normal controls ( P<0.05). Elevated EFHD2 expression correlated with advanced clinical stage ( P<0.05) and poor differentiation ( P<0.05). In the CCK-8 assay, the EFHD2 overexpression group demonstrated significantly higher cell viability than the control group, as evidenced by 450 nm relative absorbance values on Day 1 (0.529±0.019 vs. 0.515±0.016, F=0.041, P=0.320), Day 2 (1.356±0.019 vs. 1.094±0.042, F=3.833, P<0.001), Day 3 (2.817±0.049 vs. 2.143±0.124, F=3.833, P<0.001), and Day 4 (3.848±0.015 vs. 3.430±0.021, F=0.469, P<0.001). The EFHD2 knockdown group showed reduced cell viability compared to controls: Day 1 (0.541±0.020 vs. 0.552±0.015, F=0.098, P=0.423), Day 2 (1.154±0.009 vs. 1.326±0.029, F=2.485, P<0.001), Day 3 (2.453±0.041 vs. 2.653±0.031, F=0.479, P<0.001), and Day 4 (3.685±0.038 vs. 3.836±0.021, F=6.804, P<0.001). In colony formation assays, the overexpression group displayed a significant increase in colony numbers (254.667±23.861 vs. 186.000±16.703, F=0.865, P=0.015), whereas the knockdown group exhibited decreased colony formation (229.000±24.637 vs. 306.667±36.501, F=0.988, P=0.038). In Transwell assays, the EFHD2 overexpression group revealed enhanced migratory capacity [ (605.000±72.670) cells vs. (472.667±28.095) cells, F=2.462, P=0.042] and invasive potential [(767.333±21.221) cells vs. (414.333±16.623) cells, F=0.331, P<0.001]. The knockdown group showed attenuated migration [(311.000±71.084) cells vs. (479.667±50.846) cells, F=0.718, P=0.029] and invasion [(247.667±48.263) cells vs. (345.667±32.130) cells, F=0.727, P=0.043] compared to controls. The network of EFHD2-interacting proteins was further constructed by the STRING database, and the GO and KEGG analysis were used to perform bioinformatics analysis reveal that EFHD2 is mainly involved in actin cytoskeleton regulation. In conclusion, EFHD2 is highly expressed in HCC and is involved in the process of proliferation, migration and invasion of HCC.

This study investigates the expression pattern and functional significance of EF-hand domain-containing protein 2 (EFHD2) in hepatocellular carcinoma (HCC), with particular focus on its regulatory effects on tumor proliferation, migration, and invasion. Cellular experimental study was completed from June 2024 to January 2025 in the Basic Laboratory of the General Hospital of Southern Theater Command. TCGA database to determine EFHD2 expression and its clinicopathological correlations. GSCA database to assess methylation patterns and immune infiltration. Model of transient overexpression and knockdown of EFHD2 was constructed in hepatocellular carcinoma cells Hep3B, then RT-qPCR and Western blot were applied to verify the transfection efficiency. CCK-8 and colony formation assays for proliferation assessment, Transwell chambers for migration/invasion quantification. Protein-protein interaction networks were constructed via STRING, followed by GO/KEGG enrichment analysis. Statistical analysis was performed using the two independent samples t-test. The results showed that EFHD2 demonstrated significant upregulation in HCC tissues versus normal controls ( P<0.05). Elevated EFHD2 expression correlated with advanced clinical stage ( P<0.05) and poor differentiation ( P<0.05). In the CCK-8 assay, the EFHD2 overexpression group demonstrated significantly higher cell viability than the control group, as evidenced by 450 nm relative absorbance values on Day 1 (0.529±0.019 vs. 0.515±0.016, F=0.041, P=0.320), Day 2 (1.356±0.019 vs. 1.094±0.042, F=3.833, P<0.001), Day 3 (2.817±0.049 vs. 2.143±0.124, F=3.833, P<0.001), and Day 4 (3.848±0.015 vs. 3.430±0.021, F=0.469, P<0.001). The EFHD2 knockdown group showed reduced cell viability compared to controls: Day 1 (0.541±0.020 vs. 0.552±0.015, F=0.098, P=0.423), Day 2 (1.154±0.009 vs. 1.326±0.029, F=2.485, P<0.001), Day 3 (2.453±0.041 vs. 2.653±0.031, F=0.479, P<0.001), and Day 4 (3.685±0.038 vs. 3.836±0.021, F=6.804, P<0.001). In colony formation assays, the overexpression group displayed a significant increase in colony numbers (254.667±23.861 vs. 186.000±16.703, F=0.865, P=0.015), whereas the knockdown group exhibited decreased colony formation (229.000±24.637 vs. 306.667±36.501, F=0.988, P=0.038). In Transwell assays, the EFHD2 overexpression group revealed enhanced migratory capacity [ (605.000±72.670) cells vs. (472.667±28.095) cells, F=2.462, P=0.042] and invasive potential [(767.333±21.221) cells vs. (414.333±16.623) cells, F=0.331, P<0.001]. The knockdown group showed attenuated migration [(311.000±71.084) cells vs. (479.667±50.846) cells, F=0.718, P=0.029] and invasion [(247.667±48.263) cells vs. (345.667±32.130) cells, F=0.727, P=0.043] compared to controls. The network of EFHD2-interacting proteins was further constructed by the STRING database, and the GO and KEGG analysis were used to perform bioinformatics analysis reveal that EFHD2 is mainly involved in actin cytoskeleton regulation. In conclusion, EFHD2 is highly expressed in HCC and is involved in the process of proliferation, migration and invasion of HCC.

This study aimed to investigate the differences in peripheral blood lymphocyte subsets among patients with different immune statuses in the early postoperative period after liver transplantation, as well as the dynamic changes during the early post-transplantation period. A retrospective study was conducted, selecting a total of 82 patients who underwent liver transplantation at the General Hospital of PLA Southern Theater Command from January, 2018 to December, 2023. Based on the patients′ postoperative immune status, they were categorized into stable group ( n=40), infection group ( n=21), and rejection group ( n=21). Peripheral blood samples of 2-3 ml were collected from patients at weeks 1 to 4 postoperatively, and flow cytometry was employed to measure the absolute values of peripheral blood lymphocyte subsets. For metric data conforming to normal distribution and homogeneity of variance, multiple group comparisons were conducted using ANOVA and Bonferroni multiple comparisons; for non-normally distributed data, the Kruskal Wallis test was used. Friedman test was used to compare different time periods within 4 weeks after liver transplantation. The results showed that there were no statistically significant differences in the absolute values of lymphocyte subsets among the three groups in the first week after liver transplantation ( P>0.05); however, significant differences were observed in the absolute values of lymphocyte subsets among the three groups in the second, third, and fourth weeks postoperatively ( P<0.05). In the second week, the rejection group showed significantly higher absolute counts of T cells, CD4 +T cells, CD8 +T cells, NK cells, and B cells compared to the infection group (585.0 vs. 199.0; 324.0 vs.113.0; 188.0 vs.56.0; 57.0 vs.11.0; 145.0 vs.65.0 cells/μl), with statistically significant differences ( Z=-3.972, P<0.001; Z=-3.590, P=0.001; Z=-3.978, P<0.001; Z=-3.072, P=0.006; Z=-2.472, P=0.040). In the third week, the rejection group showed significantly higher absolute counts of T cells, CD4 +T cells, and CD8 +T cells compared to the infection group (660.0 vs.216.0; 350.0 vs.123.0; 184.0 vs.76.0 cells/μl), with statistically significant differences ( Z=-3.019, P=0.008; Z=-3.492, P=0.001; Z=-2.845, P=0.013). In the fourth week, the rejection group showed significantly higher absolute counts of T cells, CD4 +T cells, CD8 +T cells, and B cells compared to the infection group (690.0 vs.273.0; 405.0 vs.168.0; 214.0 vs.96.0; 117.0 vs.48.0 cells/μl), with statistically significant differences ( Z=-3.379, P=0.002; Z=-3.068, P=0.006; Z=-3.007, P=0.0086; Z=-2.330, P=0.020). Within 4 weeks after liver transplantation, the absolute values of T cells, CD8 +T cells, and NK cells in the fourth week were higher than those in the first week, with statistically significant differences ( Z=-3.825, P=0.001; Z=-3.466, P=0.003; Z=-3.526, P=0.003); however, the absolute values of B cells showed an overall decreasing trend, and were significantly lower in the fourth week than in the first and second weeks, with statistically significant differences ( Z=3.705, P=0.001; Z=2.630, P=0.009). The changes in lymphocyte subset absolute values in the rejection group were more significant than those in the infection group, with T cells, CD4 +T cells, and CD8 +T cells showing significant increases in the second, third, and fourth weeks postoperatively compared with the first week, with statistically significant differences ( Z=-3.466, P=0.003; Z=-4.661, P<0.001; Z=-5.020, P<0.001; Z=-2.749, P=0.036; Z=-4.422, P<0.001; Z=-4.542, P<0.001; Z=-3.466, P=0.003; Z=-3.765, P=0.001; Z=-4.482, P<0.001); NK cell absolute values in the third and fourth weeks postoperatively were significantly higher than those in the first week, with statistically significant differences ( Z=-2.570, P=0.061; Z=-3.765, P=0.001). In summary, monitoring the differences and dynamic changes of lymphocyte subsets in patients after liver transplantation may have certain guiding significance for evaluating the immune function status of patients and adjusting treatment plans.

This study aimed to investigate the differences in peripheral blood lymphocyte subsets among patients with different immune statuses in the early postoperative period after liver transplantation, as well as the dynamic changes during the early post-transplantation period. A retrospective study was conducted, selecting a total of 82 patients who underwent liver transplantation at the General Hospital of PLA Southern Theater Command from January, 2018 to December, 2023. Based on the patients′ postoperative immune status, they were categorized into stable group ( n=40), infection group ( n=21), and rejection group ( n=21). Peripheral blood samples of 2-3 ml were collected from patients at weeks 1 to 4 postoperatively, and flow cytometry was employed to measure the absolute values of peripheral blood lymphocyte subsets. For metric data conforming to normal distribution and homogeneity of variance, multiple group comparisons were conducted using ANOVA and Bonferroni multiple comparisons; for non-normally distributed data, the Kruskal Wallis test was used. Friedman test was used to compare different time periods within 4 weeks after liver transplantation. The results showed that there were no statistically significant differences in the absolute values of lymphocyte subsets among the three groups in the first week after liver transplantation ( P>0.05); however, significant differences were observed in the absolute values of lymphocyte subsets among the three groups in the second, third, and fourth weeks postoperatively ( P<0.05). In the second week, the rejection group showed significantly higher absolute counts of T cells, CD4 +T cells, CD8 +T cells, NK cells, and B cells compared to the infection group (585.0 vs. 199.0; 324.0 vs.113.0; 188.0 vs.56.0; 57.0 vs.11.0; 145.0 vs.65.0 cells/μl), with statistically significant differences ( Z=-3.972, P<0.001; Z=-3.590, P=0.001; Z=-3.978, P<0.001; Z=-3.072, P=0.006; Z=-2.472, P=0.040). In the third week, the rejection group showed significantly higher absolute counts of T cells, CD4 +T cells, and CD8 +T cells compared to the infection group (660.0 vs.216.0; 350.0 vs.123.0; 184.0 vs.76.0 cells/μl), with statistically significant differences ( Z=-3.019, P=0.008; Z=-3.492, P=0.001; Z=-2.845, P=0.013). In the fourth week, the rejection group showed significantly higher absolute counts of T cells, CD4 +T cells, CD8 +T cells, and B cells compared to the infection group (690.0 vs.273.0; 405.0 vs.168.0; 214.0 vs.96.0; 117.0 vs.48.0 cells/μl), with statistically significant differences ( Z=-3.379, P=0.002; Z=-3.068, P=0.006; Z=-3.007, P=0.0086; Z=-2.330, P=0.020). Within 4 weeks after liver transplantation, the absolute values of T cells, CD8 +T cells, and NK cells in the fourth week were higher than those in the first week, with statistically significant differences ( Z=-3.825, P=0.001; Z=-3.466, P=0.003; Z=-3.526, P=0.003); however, the absolute values of B cells showed an overall decreasing trend, and were significantly lower in the fourth week than in the first and second weeks, with statistically significant differences ( Z=3.705, P=0.001; Z=2.630, P=0.009). The changes in lymphocyte subset absolute values in the rejection group were more significant than those in the infection group, with T cells, CD4 +T cells, and CD8 +T cells showing significant increases in the second, third, and fourth weeks postoperatively compared with the first week, with statistically significant differences ( Z=-3.466, P=0.003; Z=-4.661, P<0.001; Z=-5.020, P<0.001; Z=-2.749, P=0.036; Z=-4.422, P<0.001; Z=-4.542, P<0.001; Z=-3.466, P=0.003; Z=-3.765, P=0.001; Z=-4.482, P<0.001); NK cell absolute values in the third and fourth weeks postoperatively were significantly higher than those in the first week, with statistically significant differences ( Z=-2.570, P=0.061; Z=-3.765, P=0.001). In summary, monitoring the differences and dynamic changes of lymphocyte subsets in patients after liver transplantation may have certain guiding significance for evaluating the immune function status of patients and adjusting treatment plans.

Objective To explore the expression level of serum long non-coding RNA(lncRNA)T342620 in patients with hepatocellular carcinoma(HCC)and the clinical value of single or combined detection with al-pha-fetoprotein(AFP)for HCC.Methods Case-control studies were conducted.A total of 69 patients with primary hepatocellular carcinoma(HCC group),32 patients with hepatitis B(hepatitis B group),20 patients with liver cirrhosis(liver cirrhosis group),30 patients after transcatheter hepatic arterial chemoembolization(TACE)for primary hepatocellular carcinoma(HCC postoperative group)and 50 healthy patients(health ex-amination group)treated in the General Hospital of Southern Theatre Command of PLA from April 2021 to May 2023 were selected as the study objects.The serum total RNA was extracted and the relative expression level of lncRNA T342620 in serum was detected by real-time quantitative PCR.Combined with the clinical di-agnosis and treatment data of patients,the correlation between its expression and pathological characteristics and serological indexes was analyzed,and the specificity and sensitivity of lncRNA T342620 alone and in com-bination with AFP in the diagnosis of HCC were analyzed by receiver operating characteristic(ROC)curve.The diagnostic efficacy was judged according to the area under the curve,and its application value in the diag-nosis of HCC was evaluated.The chi-square test was used for comparison between groups,and the Spearman method was used for correlation analysis.Results The serum expression levels of lncRNA T342620 in liver cancer group and postoperative liver cancer group were higher than those in healthy physical examination group,hepatitis B group and liver cirrhosis group,and the differences were statistically significant(P＜0.001).Clinical pathological and serological index analysis revealed that as the tumor size increased,the serum lncRNA-T342620 expression level also increased.In the HCC group,the serum lncRNA T342620 expression level was negatively correlated with albumin(ALB)and the A/G ratio(P＜0.05),while it was positively cor-related with α-L-fucosidase(AFU)and HBV-DNA(P＜0.05).In patients from the HCC postoperative group,the serum lncRNA T342620 expression level was positively correlated with total bile acid(TBA)(P＜0.05).ROC curve analysis demonstrated that when using serum lncRNA T342620 to distinguish,the sensitivi-ty and the specificity were 55.1％and 94.1％,respectively,indicating good diagnostic value.When combined with AFP detection,the sensitivity and the specificity improved to 91.3％and 91.2％,respectively,which were higher than those of individual indicators and had a superior diagnostic efficiency with area under the cuve(AUC)of 0.954 compared to AUC of AFP or lncRNA-T342620 alone(0.906,0.758),and the differ-ences were statistically significant(P＜0.05).Conclusion Serum lncRNA T342620 may be a new serological index for the auxiliary diagnosis of HCC.

Objective : To study the guidance effects of fibroblast growth factor 3 ( FGF3) on the prethalamic γ⁃aminobutyric acid (GABA) inhibitory axons, and to explore the effect of FGF3 on the formation of thalamic axonal network. Methods : Expression and localization of FGF3 and prethalamic GABA marker gene in chicken embryonic diencephalon were detected by immunofluorescence. Early orientation of GABA axons was tracked by DiI. Three⁃dimensional gel co⁃culture experiment was carried out to investigate guidance effects of FGF3 on early prethalamic GABA inhibitory axons. Results : Prethalamic GABA cells were adjacent to the FGF3 + hypothalamas, located above the hypothalamus. DiI tracing experiments revealed that prethalamic GABA inhibitory axons had already extended into the thalamus at E6. Compared with the blank control group, FGF3 not only significantly promoted the growth of prethalamic GABA inhibitory nerve fibers in the prethalamus, but also repelled the newborn prethalamic axons to the dorsal thalamus. The number of prethalamic axons was significantly less in proximal section (towards FGF3 beads) than that in distal section (away from FGF3 beads) (P < 0. 01) . Moreover, the guidance effects of FGF3 on prethalamic axons could be blocked by the FGF pathway inhibitor SU5402. Conclusion FGF3, an axon guidance molecule expressed in hypothalamus, exerts guidance effects on the pathway selection of adjacent prethalamic GABA inhibitory nerve fibers.

Objective To investigate the differences of hand and foot morphology and genetic phenotypic characteristics in adults of Hui and Han nationality in Southwestern Henan.Methods The indicators of height,weight,hand and foot were measured by the morphological measurements,the hand and foot genetic phenotype classification was observed and performed the statistical analysis.Results The hand width,foot length and foot width of the Hui adult men and women in Southwestern Henan were(8.27±0.55,23.10±1.20,9.34±0.83)cm and(7.41±0.44,20.50±1.23,8.79±0.69) cm,respectively,while which of the Han adult men and women were(8.56±0.09,24.57±1.33,9.47±0.70)cm and(7.74±0.36,22.46±1.21,8.91±0.85) cm,respectively.The total number of both hands fingerprint ridge line in Hui adult men and women were(135.06 ± 19.87) and (125.50 ±20.44)respectively,and which in Han adult men and women were (144.46 ±14.08) and (129.20 ± 20.34)respectively,the difference was statistically significant(P＜0.05).The tPD,atd angle and a-b crest line number among the Hui and Han nationalities were 16.07± 6.46,(44.61±8.66)°,34.04±5.47 and 16.53±6.27,(43.19±9.52)°,36.73±4.22 respectively.And the handedness,fingernail form,thumb type,footedness,right type ratio of foot and toe length of the Hui and Han nationalities were 90.01％,38.52％,85.59％,70.47％,56.92％ and 89.33％,45.26％,70.91％,96.98％,74.89％,respectively,the difference between the Hui and Han nationalities was statistically significant(P＜0.05).Conclusion The national differences and gender differences exist in the multiple indicators of hand and foot morphology,finger and palm prints,and genetic phenotype among the Hui and Han adults in Southwestern Henan.

Objective CRISPR/Cas9 genome-editing technique provides an novel method for whole genome editing in eukaryotic cells.Recently,we found that gene subtype library with smaller size and focused pur-pose is more economical and practical. In this study,we aimed to target kinases,a group of pivotal cell signal transducers,to construct a kinase knock-out library using CRISPR/Cas9 technique.The construction strategy wll al-so be discussed. Methods 10 sgRNA was designed for each kinase target.After oligo pool synthesis by semicon-ductor chip,the oligos were eluted from the chip. The oligo templates were amplified and cloned into Cas9 vector and transformed into Stble3 competent cells.Monoclonal colonies were selected for DNA sequencing. Results(1) GO analysis of 507 cell kinases showed that the cell kinases took part in a wide range of cell signaling.(2)The sgRNA pool with about 140 bp in length was successfully amplified by using oligo pool as the template and univer-sal PCR primers.(3)In 40 identified library clones,34 clones were sequenced successfully. Among them,the DNA sequencing results of 25 samples were completely consistent with the designed target sequences.But there are some mutations in the primers of 9 samples.Failure in bacteria shaking,DNA sequencing and other factors were ex-isted in the other clones. Conclusion The CRISPR/Cas9 kinase knock-out library can be widely used for screen-ing the important kinases which may mediate cell proliferation,metastasis,drug resistance and autophagy.This li-brary will play an important role in clarifying the development of disease associated with kinases.

OBJECTIVETo analyze and evaluate the clinical efficacy of heat-sensitive moxibustion for symptoms of large intestine cancer.

METHODSSixty patients with large intestine cancer were randomly divided into an observation group and a control group, 30 cases in each one. FOLFOX chemotherapy regimen was used in the two groups,and heat-sensitive moxibustion was added in the observation group. The acupoints were Zusanli(ST 36), Sanyinjiao (SP 6) Xuehai (SP 10) and Geshu (BL 17), etc. The treatment was applied once a day,five-day treatment as one course. Four courses were required. The reaction rates of uncomfortable symptoms by the Chinese version of the M. D. Anderson symptom inventory (MDASI-C) scale and clinical effects were analyzed and evaluated in the two groups.

RESULTSAfter treatment, the MDASI-C reaction rate of uncomfortable symptoms in the observation group was 50.4% which was lower than 53.3% in the control group (P < 0.05). The total effective rate of symptom improvement in the observation group was 83.3% (25/30), which was higher than 60.0% (18/30) in the control group (P < 0.05).

CONCLUSIONHeat-sensitive moxibustion can improve symptoms of chemotherapy for large intestine cancer.

Aged ; Antineoplastic Agents ; adverse effects ; therapeutic use ; Antineoplastic Combined Chemotherapy Protocols ; adverse effects ; therapeutic use ; Colorectal Neoplasms ; drug therapy ; Drug-Related Side Effects and Adverse Reactions ; etiology ; therapy ; Female ; Fluorouracil ; adverse effects ; therapeutic use ; Humans ; Intestine, Large ; drug effects ; Leucovorin ; adverse effects ; therapeutic use ; Male ; Middle Aged ; Moxibustion ; instrumentation ; Organoplatinum Compounds ; adverse effects ; therapeutic use ; Treatment Outcome