1.Discrimination of half-sibling relationship using diverse reference samples
Haixia LI ; Yuluo LIU ; Xuegang LI ; Huidan LIAO
Chinese Journal of Forensic Medicine 2025;40(3):369-372,374
Objective To analyze 25 cases of the half-sibling identification involving two mothers(same father,different mothers)by using family reconstruction method.Methods Hypothesis testing for half-sibling relationships were established using likelihood ratio methods.The family reconstruction was performed according to the rule of Mendelian inheritance.HSI values were calculated for dyads(two individuals),triads(three individuals with one biological mother),and tetrads(four individuals with two biological mothers)along with unrelated control groups,using 23 and 41 autosomal STR markers.Results Compared to the dyadic and triadic groups,the tetrad group exhibited a significant increase in HSI values(102~105),enabling clear discrimination of half-sibling relationships.Overlapping areas decreased,from 20.29%to 0.83%,which was reduced by 24 times,indicating enhanced system efficacy.Conclusion Incorporating additional reference samples is a feasible strategy to improve the efficacy of half-sibling identification under constrained analytical conditions.The approach enhances the detection reliability and offers valueable insights for forensic genetic practice.
2.Down-regulation of CD151 combined with bevacizumab inhibits the growth and microvessel density of colorectal cancer
Yancai LIU ; Xuegang LIU ; Zhenya ZHANG
Basic & Clinical Medicine 2025;45(1):20-24
Objective To investigate the effects of CD151 down-regulation combined with bevacizumab on colorectal cancer growth and microvessel density.Methods Human colorectal cancer cell line HT-29 and CD151--HT-29 cells strain(CD151 down-regulated HT-29 cells)were treated with bevacizumab.The cells were divided into four groups:control(HT-29)group,bevacizumab-treatment group,CD151--HT-29 group,and CD151--HT-29+bevacizumab-treatment group.Cell proliferation was observed in each group using the MTS assay.A subcutaneous xenograft model in nude mice was established,and the HT-29 control group and CD151--HT-29 group were treated with either 0.9%NaCl solution or bevacizumab.The growth of subcutaneous tumors in the four groups was ob-served,and the volume and weight of the tumors were recorded.Tumor tissues were collected for immunohistochem-ical staining of endothelial cells to assess microvessel density(MVD).Results Compared with the control group,cell proliferation was significantly reduced in the bevacizumab-treated group and CD151--HT-29 group(P<0.001).Cell proliferation in the CD151--HT-29+bevacizumab-treated group was slower than that in the single treatment groups(P<0.001).In the subcutaneous tumor model,the volume,weight,and MVD of tumors in the bevacizumab-treated and CD151--HT-29 groups were significantly reduced compared to the control group(P<0.01).In the CD151--HT-29+bevacizumab group,the tumor volume,weight,and CD34 expression were significantly lower than in the single treatment groups(P<0.01).Conclusions CD151 protein may play a role in the regulation of angiogenesis in colorectal cancer tissues and may have a synergistic effect with bevacizumab in in-hibiting microvessel formation in tumor tissues.
3.Application of holographic imaging digital localization technology in robotic partial nephrectomy for completely endophytic renal tumor
Zhengsheng LIU ; Shuaishuai SONG ; Xuegang WANG ; Kaiyan ZHANG ; Wei LI ; Zhun WU ; Bin CHEN ; Tao WANG ; Jinchun XING
Chinese Journal of Urology 2025;46(5):376-382
Objective:To evaluate the clinical value of holographic imaging-based digital localization technology in robot-assisted partial nephrectomy(RAPN)for the treatment of completely endophytic renal tumors.Methods:A retrospective analysis was conducted on the clinical data of 23 patients with completely endophytic renal tumors who underwent RAPN at the First Affiliated Hospital of Xiamen University between December 2022 and December 2024. Patients were divided into two groups based on the use of holographic imaging:the holographic imaging group(16 cases)and the conventional group(7 cases). There was no statistically significant difference between the holographic imaging group and the conventional group in terms of age[(41.9 ± 13.4)years vs.(46.9 ± 13.4)years],body mass index[(25.6 ± 4.8)kg/m2 vs.(24.7 ± 3.1)kg/m2],maximum tumor diameter[(3.1 ± 0.9)cm vs.(3.0 ± 9.0)cm],tumor volume[(13.2 ± 9.0)cm3 vs.(34.9 ± 9.9)cm3],R.E.N.A.L. score[(9.4 ± 1.2)points vs.(9.9 ± 0.7)points],PADUA score[(10.4 ± 0.7)points vs.(9.4 ± 0.7)points],proportion of T 1a stage patients[12 cases(75.0%)vs. 6 cases(85.7%)]and preoperative serum creatinine[(67.4 ± 9.5)μmol/L vs.(78.0 ± 16.0)μmol/L]. In the holographic group,holographic models were reconstructed based on preoperative enhanced CT or MRI images and used for preoperative planning and intraoperative localization. In the conventional group,surgeons relied on preoperative CT or MRI images for cognitive fusion during RAPN. Perioperative parameters such as warm ischemia time,operative time,tumor localization time,positive surgical margin rate,and renal function were compared between the two groups. Results:The operative time in the holographic group was significantly shorter than that in the conventional group[(152.8 ± 12.9)min vs.(218.4 ± 105.5)min, P = 0.001]. Warm ischemia time[(26.9 ± 3.4)min vs.(30.7 ± 3.8)min, P < 0.001],localization time[(4.2 ± 0.9)min vs.(8.9 ± 1.7)min, P < 0.001],and estimated blood loss[(47.0 ± 17.7)ml vs.(128.6 ± 87.8)ml, P < 0.001]were also significantly lower in the holographic group. In the conventional group,one patient underwent radical nephrectomy,while no patient in the holographic imaging group required conversion to radical nephrectomy. No cases of positive surgical margins were identified in either group. Serum creatinine levels measured one month after surgery showed no statistically significant differences between the two groups[(79.5 ± 15.7)μmol /L vs.(104.9 ± 22.5)μmol /L]. Conclusions:The application of holographic imaging-based digital localization technology in RAPN for completely endophytic renal tumors significantly reduces operative time,localization time,warm ischemia time,and intraoperative blood loss. This technology improves surgical efficiency and success rates,offering distinct clinical advantages.
4.Discrimination of half-sibling relationship using diverse reference samples
Haixia LI ; Yuluo LIU ; Xuegang LI ; Huidan LIAO
Chinese Journal of Forensic Medicine 2025;40(3):369-372,374
Objective To analyze 25 cases of the half-sibling identification involving two mothers(same father,different mothers)by using family reconstruction method.Methods Hypothesis testing for half-sibling relationships were established using likelihood ratio methods.The family reconstruction was performed according to the rule of Mendelian inheritance.HSI values were calculated for dyads(two individuals),triads(three individuals with one biological mother),and tetrads(four individuals with two biological mothers)along with unrelated control groups,using 23 and 41 autosomal STR markers.Results Compared to the dyadic and triadic groups,the tetrad group exhibited a significant increase in HSI values(102~105),enabling clear discrimination of half-sibling relationships.Overlapping areas decreased,from 20.29%to 0.83%,which was reduced by 24 times,indicating enhanced system efficacy.Conclusion Incorporating additional reference samples is a feasible strategy to improve the efficacy of half-sibling identification under constrained analytical conditions.The approach enhances the detection reliability and offers valueable insights for forensic genetic practice.
5.Application of holographic imaging digital localization technology in robotic partial nephrectomy for completely endophytic renal tumor
Zhengsheng LIU ; Shuaishuai SONG ; Xuegang WANG ; Kaiyan ZHANG ; Wei LI ; Zhun WU ; Bin CHEN ; Tao WANG ; Jinchun XING
Chinese Journal of Urology 2025;46(5):376-382
Objective:To evaluate the clinical value of holographic imaging-based digital localization technology in robot-assisted partial nephrectomy(RAPN)for the treatment of completely endophytic renal tumors.Methods:A retrospective analysis was conducted on the clinical data of 23 patients with completely endophytic renal tumors who underwent RAPN at the First Affiliated Hospital of Xiamen University between December 2022 and December 2024. Patients were divided into two groups based on the use of holographic imaging:the holographic imaging group(16 cases)and the conventional group(7 cases). There was no statistically significant difference between the holographic imaging group and the conventional group in terms of age[(41.9 ± 13.4)years vs.(46.9 ± 13.4)years],body mass index[(25.6 ± 4.8)kg/m2 vs.(24.7 ± 3.1)kg/m2],maximum tumor diameter[(3.1 ± 0.9)cm vs.(3.0 ± 9.0)cm],tumor volume[(13.2 ± 9.0)cm3 vs.(34.9 ± 9.9)cm3],R.E.N.A.L. score[(9.4 ± 1.2)points vs.(9.9 ± 0.7)points],PADUA score[(10.4 ± 0.7)points vs.(9.4 ± 0.7)points],proportion of T 1a stage patients[12 cases(75.0%)vs. 6 cases(85.7%)]and preoperative serum creatinine[(67.4 ± 9.5)μmol/L vs.(78.0 ± 16.0)μmol/L]. In the holographic group,holographic models were reconstructed based on preoperative enhanced CT or MRI images and used for preoperative planning and intraoperative localization. In the conventional group,surgeons relied on preoperative CT or MRI images for cognitive fusion during RAPN. Perioperative parameters such as warm ischemia time,operative time,tumor localization time,positive surgical margin rate,and renal function were compared between the two groups. Results:The operative time in the holographic group was significantly shorter than that in the conventional group[(152.8 ± 12.9)min vs.(218.4 ± 105.5)min, P = 0.001]. Warm ischemia time[(26.9 ± 3.4)min vs.(30.7 ± 3.8)min, P < 0.001],localization time[(4.2 ± 0.9)min vs.(8.9 ± 1.7)min, P < 0.001],and estimated blood loss[(47.0 ± 17.7)ml vs.(128.6 ± 87.8)ml, P < 0.001]were also significantly lower in the holographic group. In the conventional group,one patient underwent radical nephrectomy,while no patient in the holographic imaging group required conversion to radical nephrectomy. No cases of positive surgical margins were identified in either group. Serum creatinine levels measured one month after surgery showed no statistically significant differences between the two groups[(79.5 ± 15.7)μmol /L vs.(104.9 ± 22.5)μmol /L]. Conclusions:The application of holographic imaging-based digital localization technology in RAPN for completely endophytic renal tumors significantly reduces operative time,localization time,warm ischemia time,and intraoperative blood loss. This technology improves surgical efficiency and success rates,offering distinct clinical advantages.
6.Mechanism of glioma stem cells with high expression of PTPRZ1 inducing TAMs polarization to M2 immunosuppressive phenotype
Lele AN ; Ying YANG ; Qing LIU ; Feiyue DOU ; Lujing WANG ; Yue CHENG ; Chao WANG ; Qianying RUAN ; Lei ZHOU ; Haitao GUO ; Weikai KONG ; Xuegang LI ; Chuan LAN ; Fei LI ; Yu SHI
Journal of Army Medical University 2024;46(8):796-803
Objective To explore the effect of glioma stem cells with high expression of protein tyrosin phosphatase receptor type Z1 (PTPRZ1 )on the phenotypic polarization and phagocytosis of tumor-associated macrophages and its regulatory mechanism.Methods GSCs and non-stem tumor cells (NSTCs) were screened out from human glioblastoma (GBM) specimens using flow cytometry,and the PTPRZ1 expression in paired GSCs and NSTCs were detected.Human peripheral blood mononuclear cells (PBMC)-derived CD14+monocytes were exposed to the conditioned medium from glioma cells or recombinant chemokine C-C motif ligand 20 (CCL20)for TAM polarization.Stable PTPRZ1 knockout GSCs (PTPRZ1-KO GSCs) were constructed using CRISPR/Cas9. TAM phagocytosis to GSCs,NSTCs,PTPRZ1-Control GSCs (PTPRZ1-Ctrl GSCs)and PTPRZ1-KO GSCs and the expression of immunosuppressive phenotype (M2) polarization marker CD163 were examined using flow cytometry.Differentially expressed genes (DEGs ) between paired GSCs and NSTCs were determined using a bulk RNA-sequencing dataset (GSE54791 )from Gene Expression Omnibus (GEO).A gene set informing worse outcome of patients with GBM was generated using The Cancer Genome Atlas (TCGA)-GBM cohort.By intersecting the aforementioned gene set with the gene set that encodes for human membrance proteins,the PTPRZ1 gene is obtained.Gene set enrichment analysis (GSEA)was used for pathway enrichment analysis to compare the differentially regulated pathways between GBMs with high or low PTPRZ1 expression.Bulk RNA sequencing,qRT-PCR and Western blotting were used to identify the DEGs between PTPRZ1-KO GSCs and PTPRZ1-Ctrl GSCs.Results GSCs were more capable of escaping from TAM phagocytosis than NSTCs (P<0.05 )and had specifically up-regulated PTPRZ1 expression.PTPRZ1-KO significantly suppressed GSCs escaping from TAM phagocytosis (P<0.01 ). GBMs with high PTPRZ1 expression showed significant inhibition of pathways mediating phagocytosis (P<0.05).The expression of CCL20 as a M2 TAM polarization chemokine was significantly down-regulated in PTPRZ1-KO GSCs (P<0.05 ).Treatment with recombinant CCL20 up-regulated the expression of CD163 as a M2 TAM marker in TAM.Conclusion PTPRZ1+GSCs mediate M2 TAM polarization and inhibit TAM phagocytosis,which may be related to the up-regulation of CCL20 in PTPRZ1+GSCs.
7.Genetic engineering and molecular modification of recombinant fully humanized single-domain antibody against Helicobacter pylori UreB
Xuefang WANG ; Yang ZHAO ; Zhuqing LIU ; Le GUO ; Feiliang ZHONG ; Xuegang LUO
Journal of China Pharmaceutical University 2024;55(5):666-672
To construct a recombinant expression system for a single-domain antibody targeting the urease of Helicobacter pylori(Hp),this study employed several strategies.First,using artificial intelligence(AI)auxiliary tools such as Pymol,I-TASSER,and ClussPro2,the molecular interactions between different antibodies and Hp urease subunit B(UreB)were analyzed.The fully humanized single-domain antibody UreBAb was identified as the primary research target.Next,the UreBAb gene sequence was optimized based on Escherichia coli codon preferences,and was inserted into expression vectors such as pET28a and pE-SUMO.The resulting recombinant expression strains were obtained by transforming Escherichia coli Rosetta(DE3).Recombinant antibody proteins were prepared through IPTG induction,and its activity was detected using extracted Hp urease as the antigen.SDS-PAGE analysis confirmed the correct expression of both UreBAb and SUMO-UreBAb,with protein yields of 0.34 mg/mL and 0.41 mg/mL,respectively.Unidirectional immunodiffusion experiments further confirmed that both recombinant antibodies exhibited strong affinity for Hp UreB antigen,with inhibition rates of 51.27%and 74.07%,respectively.Additionally,leveraging artificial intelligence tools such as AlphaFold2,cluspro2,mCSM-AB,OSPREY,and FoldX,the study evaluated and analyzed key binding sites and mutational strategies affecting the stability of the antigen-antibody complex.Subsequently,nine UreBAb evolution mutants were constructed,and their binding activities with the antigen were enhanced.Among these,the I107W mutant showed the most significant improvement,achieving a 24.95%increase compared to the wild-type UreBAb.This research lays a solid foundation for the development of fully humanized single-domain antibodies against Hp.
8.Human umbilical cord mesenchymal stem cell-derived extracellular vesicles enhance the regenerative capability of fibrotic liver
Yunguo LEI ; Jia YAO ; Jun ZHENG ; Tongyu LU ; Jiebin ZHANG ; Jiaqi XIAO ; Yasong LIU ; Haitian CHEN ; Xuegang ZHAO ; Xingye YANG
Organ Transplantation 2023;14(3):379-
Objective To investigate the role of human umbilical cord mesenchymal stem cell-derived extracellular vesicle (hUC-MSC-EV) in the regeneration of fibrotic liver. Methods C57BL/6 mice were randomly divided into the 70% normal liver resection group (Oil+PHx group), 70% liver fibrosis resection group (CCl4+PHx group) and 70% liver fibrosis resection+mesenchymal stem cell-derived extracellular vesicle (MSC-EV) treatment group (CCl4+PHx+MSC-EV group), with 8 mice in each group. LX-2 cell lines were assigned into the phosphate buffer solution (PBS) group, transforming growth factor (TGF)-β group and TGF-β+MSC-EV group. The serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and lactate dehydrogenase (LDH) in mice after partial liver resection were detected in each group. The expression levels of liver fibrosis and proliferation-related parameters were analyzed in each group. The messenger RNA (mRNA) expression levels of epidermal growth factor (EGF), fibroblast growth factor (FGF), vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF) in LX-2 cells were detected in each group, and their effects on HGF expression in mouse liver were observed. Results Compared with the Oil+PHx group, the serum levels of AST, ALT and LDH were up-regulated, and the degree of fibrosis was more severe, the positive area of Sirius red and α-smooth muscle actin (α-SMA) staining was larger, and the expression level of α-SMA protein was up-regulated in the CCl4+PHx group. Compared with the CCl4+PHx group, the serum levels of AST, ALT and LDH were decreased, the degree of fibrosis was slighter, the positive area of Sirius red and α-SMA staining was decreased, and the expression level of α-SMA protein was down-regulated in the CCl4+PHx+MSC-EV group, and the differences were statistically significant (all
9.A novel surgical difficulty scoring system for renal carcinoma based on holographic imaging
Zhengsheng LIU ; Zhun WU ; Xuegang WANG ; Kaiyan ZHANG ; Wei LI ; Chaohao MIAO ; Yu LUO ; Jinchun XING
Chinese Journal of Urology 2022;43(5):344-349
Objective:To establish surgical difficulty scoring system of partial nephrectomy based on holographic imaging and explore its application value in partial nephrectomy.Methods:A total of 184 patients including 110 males and 74 females with renal tumors diagnosed as stage cT 1 to cT 2 before surgery in the First Affiliated Hospital of Xiamen University from October 2019 to January 2022 were included. Among the 184 patients, 141 patients were treated with partial nephrectomy and 43 patients were treated with radical nephrectomy (3 partial nephrectomies were finally changed to radical nephrectomies due to vascular and tumor location). 60 patients had hypertension. 24 patients had diabetes. 7 patients had hyperuricemia. The median age was 55(47, 62) years. The median BMI was 23.7(21.8, 26.4) kg/m 2. The median maximum tumor diameter was 3.9(2.9, 5.2) cm. The median preoperative eGFR was 99.7(83.4, 114.2) ml/(min·1.73m 2). The median R. E.N.A.L. score was 8(6, 9). The median PADUA score was 9(8, 10). 153 patients were diagnosed as stage cT 1 and 31 patients were diagnosed as stage cT 2. The hologram reconstruction was performed according to preoperative CT or MRI examination. The maximum diameter of the tumor in the kidney (D), the compression degree of the renal segmental vessels by tumor(C), the area of the renal sinus occupied by tumor(O) and the mass of exophytic rate(M) were comprehensively considered and finally constituted the difficulty scoring system named DCOM score for partial nephrectomy. The DCOM score divided the complexity of tumor surgery into mild (4-6 points), moderate (7-8 points) and high (≥ 9 points). Meanwhile, the MIC (surgical margins are negative, WIT is <20 min, and no major complications)was used to evaluate the overall surgical effect. The DCOM, R. E.N.A.L. and PADUA scores were performed on all patients and compared with each other to evaluate the surgical effect of DCOM score in partial nephrectomy. Results:All surgeries in this study were successfully completed, including 141 partial nephrectomies and 43 radical nephrectomies. The DCOM score was 10(9, 11) for radical nephrectomy and 6(5, 8) for partial nephrectomy, and the difference was statistically significant ( P=0.001). There were 23 patients (37.7%) in highly complex group, 39 patients (88.6%) in moderately complex group and 79 patients (100.0%) in mildly complex group underwent partial nephrectomy, respectively. According to multifactorial analysis, patients in highly and moderately complex group of DCOM score had 8.88 times ( P=0.001) and 1.76 times ( P=0.005) less reach MIC than those in mildly complex group, respectively. Patients in highly and moderately complex group of PADUA score had 4.86 times ( P=0.005)and 3.41 times ( P=0.006)less reach MIC than patients in mildly complex group of DCOM score, respectively. What’s more, patients in moderately complex group of R. E.N.A.L. score had 3.11 times ( P=0.003) less reach MIC than patients in mildly complex group of DCOM score. In the ROC curves to predict MIC achievement, the AUC values of R. E.N.A.L., PADUA and DCOM scores were 0.657, 0.655 and 0.746, respectively. Comparing:R. E.N.A.L. score with DCOM score, the AUC value was statistically significant ( P=0.025). Conclusions The surgical difficulty scoring system (DCOM score) based on holographic imaging can predict the outcome of partial nephrectomy, but further verification is needed.
10.Unqualified rate of anti-HIV detection in the laboratories from blood banks in Beijing-Tianjin-Hebei region
Wei ZHEN ; Hongwei GE ; Rui WANG ; Tong PAN ; Wei HAN ; Peng WANG ; Li YANG ; Shaoqiu SUN ; Xiao CAO ; Liye CUI ; Chao WEI ; Guijun YU ; Yunpeng XU ; Jinjuan FANG ; Caixia LIU ; Xuegang WANG ; Zhijun ZHEN ; Xiaojie LIU ; Wengong DU ; Lunan WANG ; Jiang LIU ; Hongjie WANG
Chinese Journal of Blood Transfusion 2021;34(4):371-376
【Objective】 To investigate the unqualified rate of anti-HIV detection of blood screening laboratories in Beijing-Tianjin-Hebei region, and explore the differences in anti-HIV detection ability and influencing factors in each laboratory. 【Methods】 Through filling questionnaires via e-mail, the anti-HIV ELISA unqualified rate and confirmed (WB) positive results (data) from January to December 2018 from 15 blood screening laboratories in Beijing-Tianjin-Hebei region were collected. Our laboratory was responsible for data collection and confirmation, and statistics software SPSS22.0 was used for analysis. 【Results】 1) There was a statistically significant difference among the unqualified rate of anti-HIV ELISA(6.77‱~35.71‱) and confirmed positive rate(0.60‱~3.56‱) in 15 blood screening laboratories in Beijing-Tianjin-Hebei region (P<0.05); 2) There were significant differencse among the ELISA unqualified rate and the confirmed positive rate of 8 reagents for anti-HIV detection(P<0.01), and the sensitivity of the 4th generation detection reagent and the imported reagent was higher than that of the 3rd generation reagent and the domestic reagent. The anti-HIV ELISA unqualified rate of R5 was the highest (19.08‱). 3)There were significant differences in the anti-HIV ELISA unqualified rate of R1, R2, R3, R5 and R7 reagents among different blood station laboratories(P<0.05), and there were no significant differences in the anti-HIV ELISA unqualified rate of R4, R6 and R8 reagents among different blood station laboratories(P>0.05). 4)The unqualified rate of anti-HIV ELISA of laboratories using different regents showed significant differences(P<0.05), except H, J, M. The unqualified rate of imported reagent was significantly higher than that of domestic reagents of laboratories using imported and domestic reagents combinations(P<0.05), except O. 62.5% (5/8) laboratories using domestic 3rd and 4th generation reagent combination showed significant differences in the unqualified rates among different reagents(P<0.05); 5) The positive rate of single-reagent(62.02%~95.45%)in 15 blood screening laboratories showed significant difference(P<0.001), and A was the lowest (62.02%). 【Conclusion】 The anti-HIV detection ability among 15 blood screening laboratories in Beijing-Tianjin-Hebei region is quite different. The application of different reagents is the main factor for the difference, and other factors such as personnel, instruments and test strategies also has a great impact on the detection of anti-HIV. It is still necessary to promote the process of homogenization of blood testing quality among blood screening laboratories in Beijing-Tianjin-Hebei region.

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