1.Corporate Governance and Workplace Mental Health Practices: The Mediating Role of Structured Occupational Safety and Health Engagement
Ro-Ting LIN ; Lung-Chang CHIEN ; Chieh-Wen CHANG ; Yu-Chi LIAO ; Tomohisa NAGATA
Safety and Health at Work 2026;17(1):33-39
Background:
This study examines whether stronger corporate governance is associated with workplace mental health practices and whether this relationship is mediated by a structured sequence of occupational safety and health (OSH) engagement: recognition, goal-setting, and implementation.
Methods:
We analyzed 134 listed companies in Taiwan that published sustainability reports and received corporate governance evaluations between 2014 and 2023. Governance scores―based on shareholder rights, board functioning, transparency, and sustainability―were dichotomized into high vs. low categories. Workplace mental health practices were measured using 24 binary indicators across planning, provision, and reporting. OSH engagement was conceptualized in three stages based on Global Reporting Initiative 403 guidelines, with composite indicators derived via grouped weighted quantile sum regression. Associations were analyzed using generalized linear mixed-effects and serial mediation models.
Results:
Compared to companies ranked low in corporate governance, those ranked high had a higher likelihood of adopting mental health practices. OSH goal-setting and implementation showed significant positive associations, whereas recognition did not. Serial mediation analysis showed that 83% of the total effect of corporate governance on mental health practices was direct, while 17% was mediated through structured OSH engagement, primarily via implementation. Recognition showed a nonsignificant effect but initiated significant sequential pathways via goal-setting and implementation.Goal-setting functioned as a bridge within these chains.
Conclusion
Corporate governance plays a central role in advancing workplace mental health practices.While most effects are direct, structured OSH engagement helps translate governance priorities into sustained organizational actions that embed mental health into routine practice.
2.Expression levels of Yes-associated protein 1 and human collagen type Ⅵ alpha 1 in pancreatic cancer under hyperglycemic conditions and their impact on prognosis
Jiahua LIAO ; Xuehuan HUANG ; Ruohan LI ; Xiping TANG ; Wen QIN
Journal of Clinical Hepatology 2026;42(5):1133-1143
ObjectiveTo investigate the changes in the expression levels of Yes-associated protein 1 (YAP1) and human collagen type Ⅵ alpha 1 (COL6A1) in pancreatic cancer under hyperglycemic conditions and their association with prognosis. MethodsGEPIA database and R software were used to analyze the expression levels of YAP1 and COL6A1 and their correlation in pancreatic cancer, as well as their impact on the prognosis of patients with pancreatic cancer. The cBioPortal database was used to identify the co-expressed genes of YAP1 and COL6A1 in pancreatic cancer, and the gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed. Quantitative real-time PCR and Western blot were used to measure the mRNA and protein expression levels of YAP1 and COL6A1 in human pancreatic cancer PANC-1 cells cultured under hyperglycemic or normoglycemic conditions. A nude mouse model of pancreatic cancer with type 2 diabetes was established, and immunohistochemistry was used to measure the expression of YAP1 and COL6A1 in subcutaneous tumor tissue. Clinical specimens and data of 96 patients with pancreatic cancer were collected from January 2016 to January 2020 in the First Affiliated Hospital of Guangxi Medical University, Guangxi Medical University Cancer Hospital and Wuming Hospital Affiliated to Guangxi Medical University. Among them, 48 patients with pancreatic cancer and diabetes were enrolled as study group, and 48 patients with pancreatic cancer alone were enrolled as control group; immunohistochemistry was used to measure the expression levels of YAP1 and COL6A1 in tissue samples from both groups, and their association with clinical indicators and survival outcomes was analyzed. The t-test was used for comparison of continuous data between groups, and the chi-square test or the Mann-Whitney U test was used for comparison of categorical data between groups; the Pearson correlation analysis was used for normally distributed continuous variables, while the Spearman correlation analysis was used for discontinuous variables or non-normally distributed variables. The Kaplan-Meier method was used to plot survival curves, and the Log-rank test was used for comparison of survival rates. The Cox proportional hazards model was used to perform univariate and multivariate analyses. ResultsBoth YAP1 and COL6A1 were highly expressed in pancreatic cancer, with a strong correlation between them (r=0.85, P<0.001), and they were associated with the poor prognosis of patients (hazard ratio [HR]=1.8 and 1.5, both P<0.05). YAP1 and COL6A1 were significantly correlated with 219 and 122 genes, respectively (r=0.83 and 0.86, both P<0.05), and the GO and KEGG enrichment analyses showed that these genes affected the development and progression of pancreatic cancer through extracellular matrix-receptor interaction, the Hippo signaling pathway, and the phosphatidylinositol 3-kinase/protein kinase B signaling pathway. Quantitative real-time PCR and Western blot showed that there were significant increases in the mRNA and protein expression levels of YAP1 and COL6A1 in human pancreatic cancer PANC-1 cells under hyperglycemic conditions (mRNA expression: t=4.726 and 6.197, both P<0.05; protein expression: t=6.826 and 5.254, both P<0.05). Immunohistochemistry showed that there were significant increases in the expression levels of YAP1 and COL6A1 in subcutaneous tumor tissue of nude mice, with a positive correlation between them (r=0.985, P<0.001). For the clinical specimens of pancreatic cancer tissue, immunohistochemistry showed that there were significant increases in the positive expression levels of YAP1 and COL6A1 proteins in the study group, with a positive correlation between them (r=0.882, P<0.001). The multivariate analysis of clinical data showed that lymph node metastasis (HR=0.083, 95% confidence interval [CI]: 0.030 — 0.231, P<0.05), distant organ metastasis (HR=0.166, 95%CI: 0.065 — 0.420, P<0.05), YAP1 expression level (HR=2.027, 95%CI: 1.065 — 3.857, P<0.05), and COL6A1 expression level (HR=2.044, 95%CI: 1.019 — 4.099, P<0.05) were independent influencing factors for the prognosis of patients with pancreatic cancer. The survival analysis showed that the high YAP1 expression group had a significantly lower 2-year survival rate than the low YAP1 expression group (6.82% vs 25.00%, χ2=16.382, P<0.001), and the high COL6A1 expression group had a significantly lower 2-year survival rate than the low COL6A1 expression group (14.58% vs 18.75%, χ2=4.579, P=0.032). ConclusionThe hyperglycemic environment promotes the high expression of YAP1 and COL6A1 in pancreatic cancer, which is associated with poor prognosis in patients.
3.Analysis of specific risks and long-term toxicities of BCR-ABL1 TKIs in pediatric patients with hematological malignancies
Luping WEN ; Fan XIA ; Ziqiong LIAO ; Benjie ZHOU ; Hui CHEN
China Pharmacy 2026;37(8):1050-1055
OBJECTIVE To analyze the specific risks and long-term toxicities of four BCR-ABL1 tyrosine kinase inhibitors (TKIs)(imatinib, dasatinib, nilotinib, and bosutinib) in pediatric patients with hematological malignancies. METHODS Adverse drug event (ADE) reports submitted to the the United States FDA Adverse Event Reporting System (FAERS) from January 2012 to December 2024, with imatinib, dasatinib, nilotinib, and bosutinib as the primary suspect drugs, were collected. Data mining was performed using the reporting odds ratio method and proportional reporting ratio method. ADE terms were classified and summarized by system organ class (SOC) and preferred term (PT) according to the Medical Dictionary for Drug Regulatory Activities (MedDRA, version 26.0). Meanwhile, the ADE reports were divided by age into the adult group (≥18 years) and the pediatric group (<18 years) to compare the differences in ADE between the two groups. RESULTS A total of 1 512 pediatric ADE reports were included: 993 for imatinib, 391 for dasatinib, 112 for nilotinib, and 16 for bosutinib. Among the reported ADEs, the patients were mainly aged 12-<18 years; the reports mainly originated from the United States, France, and Japan; and the primary indications were chronic myeloid leukemia and acute lymphoblastic leukemia. A total of 5 256 ADE signals were mined, among which 235 were positive signals, involving 1 103 PT across 27 SOC. The top five PT ranked by the number of positive signals were nausea, febrile neutropenia, abdominal pain, neutropenia, and anemia. The top two SOC were general disorders and administration site conditions, and gastrointestinal disorders. Compared with the adult group, the pediatric group had relatively higher proportions of events related to infections and infestations as well as blood and lymphatic system disorders. Pediatric long-term toxicity signals primarily included growth retardation, accompanied by signals related to endocrine system abnormalities and bone metabolism abnormalities. Specific signals included imatinib-associated septic shock, dasatinib-associated chylothorax, and nilotinib-associated electrocardiographic QT interval prolongation. CONCLUSIONS When pediatric patients use BCR-ABL1 TKIs, priority monitoring of infection risk and hematologic parameters is required, along with long-term follow-up of height, endocrine, and bone metabolism parameters. Targeted screening and management of drug-specific signals should be performed to ensure the long-term safety of pediatric medication.
4.Research progress on the mechanism of action and clinical application of Shenqi dihuang decoction in the treatment of diabetic nephropathy
Jiajie LI ; Jiaqi WANG ; Jie ZHAO ; Zezhu LI ; Yaping WANG ; Guirong ZHANG ; Heguo YAN ; Jiabao LIAO ; Weibo WEN
China Pharmacy 2026;37(8):1085-1091
Diabetic nephropathy(DN) is a common and severe microvascular complication of diabetes. In recent years, the classical herbal formula Shenqi dihuang decoction has demonstrated unique advantages in the clinical treatment of DN. This article conducts a systematic review of the mechanisms of action and clinical applications of Shenqi dihuang decoction in the treatment of DN. It reveals that the mechanism by which this formula improves DN involves multi-target synergistic regulation. For instance, Shenqi dihuang decoction exerts multiple pharmacological effects by regulating signaling pathways including phosphatidy linostiol 3-kinase/protein kinase B, AMP-activated protein kinase/silent information regulator 1/forkhead box O1, and nuclear factor erythroid 2-related factor 2/heme oxygenase-1 pathways.These effects include regulating glucose and lipid metabolism, inhibiting oxidative stress, reducing inflammation, improving insulin resistance, modulating cell death (apoptosis/autophagy/ferroptosis/pyroptosis), and preventing renal fibrosis. Existing clinical studies indicate that Shenqi dihuang decoction and its modified formulas, alone or in combination with other therapeutic methods, can significantly improve glucose and lipid metabolism, reduce proteinuria, and delay renal function decline in patients with DN. These effects are superior to those of Western medicines such as irbesartan, valsartan, and empagliflozin, and the treatment demonstrates good safety. Future research should leverage systems biology and artificial intelligence technologies to further elucidate the integrated mechanisms in the treatment of DN by Shenqi dihuang decoction, thereby advancing the precision and standardization of its clinical application.
5.Prenatal diagnosis of 22q11.2 microduplication syndrome in a three-generation family: Clinical-genetic characteristics and literature review.
Yifan LIAO ; Yidong WEN ; Xiaoqin DENG ; Cimo WANG ; Zhirong SHANG ; Jinghong YANG ; Jiabing LI
Chinese Journal of Medical Genetics 2026;43(1):57-63
OBJECTIVE:
To explore the genetic etiology for a pregnant woman with a history of multiple adverse pregnancies and assess the phenotype-genotype correlation of 22q11.2 microduplication syndrome in her family.
METHODS:
Amniotic fluid sample was taken from a pregnant woman for whom non-invasive prenatal screening indicated chromosome 22 abnormalities in the fetus. Peripheral blood samples from the woman, her brother and parents were collected for high-throughput low-depth whole genome sequencing (CNV-seq). A pedigree traceability analysis of the results was conducted in conjunction with analysis of clinical manifestation. Relevant literature (from establishment to March 2025) was systematically searched. This study was approved by the Medical Ethics Committee of Mianyang Maternal and Child Health Care Hospital (Ethics No.: Lun Shen [2024]009).
RESULTS:
CNV-seq revealed that the fetus had harbored a 6.02 Mb duplication at 22q11.21q11.23. Karyotyping confirmed it as 46,X?dup(22)(q11.2). Pedigree verification demonstrated that the pregnant woman, her brother and mother had all carried the same duplication. Phenotypic analysis of the affected family members showed classic features of 22q11.2 microduplication syndrome, including hypernasal speech, low nasal bridge, congenital heart disease, and cognitive impairment. A total of 44 cases with full information (including three patients from this pedigree) were included in the analysis. The penetrance of 22q11.2 duplication was approximately 29.5% (13/44), and 52.3% (23/44) of the cases had inherited the variant from a phenotypically normal parent.
CONCLUSION
This study has identified the genetic basis for the woman's recurrent adverse pregnancies and phenotypic abnormalities in her family members. The scoliosis identified in her younger brother has not been previously reported, thereby may enrich the clinical phenotype of this syndrome. For fetuses identified with a 22q11.2 microduplication, detailed fetal imaging is recommended, and genetic counseling should be provided to the couples.
Humans
;
Female
;
Pregnancy
;
Prenatal Diagnosis/methods*
;
Chromosome Duplication/genetics*
;
Male
;
Pedigree
;
DiGeorge Syndrome/diagnosis*
;
Adult
;
Chromosomes, Human, Pair 22/genetics*
;
Abnormalities, Multiple
6.Study on the construction of a red blood cell rare blood type database and physical repository in the Guangzhou Region
Zhijian LIAO ; Shuangshuang JIA ; Yuan SHAO ; Boquan HUANG ; Chunyan MO ; Jizhi WEN ; Runqing ZHANG ; Xia RONG ; Hong LUO ; Huaqin LIANG ; Yanli JI
Chinese Journal of Blood Transfusion 2026;39(5):619-628
Objective: To conduct screening for rare blood types within important blood group systems for the Chinese population, such as Rh, Duffy, Kidd, P1Pk, Diego, and MNS, in the Guangzhou region, and to establish a corresponding rare blood type database and physical repository. Methods: The saline medium microplate method was used to screen blood donors with the ccDEE phenotype combined with either Jk(a-) or Jk(b-). The polybrene microplate method was employed to screen for donors with Fy(a-), s(-), Lu(b-), Di(b-), k(-), and p phenotypes. The urea lysis microplate method was applied to screen for the Jk(a-b-) phenotype. A high-resolution melting (HRM) curve method was established for screening some donors with the Di(b-) phenotype. Subsequently, expanded phenotyping of antigens in the Rh, Kidd, MNS, Duffy, P1Pk, Lewis, Kell, and Lutheran blood group systems was performed on identified rare blood type donors using monoclonal antibodies. The test results are entered into the Rare Blood Type Bank Management System of the Guangzhou Blood Center, enabling functions such as confirmation reminders and cryopreservation storage when the donor donates again. Red blood cells of rare blood types are processed into frozen red blood cells for long-term storage. Results: Among voluntary blood donors, 16 cases of the ccDEE combined with Jk(a-) phenotype were identified (0.221 7%, 16/7 216); 10 cases of the ccDEE combined with Jk(b-) phenotype (0.138 6%, 10/7 216); 78 cases of the Fy(a-) phenotype (0.169 5%, 78/46 012); 39 cases of the Lu(b-) phenotype (0.138 2%, 39/28 214); 31 cases of the s(-) phenotype (0.081 8%, 31/37 913); 22 cases of the Di(b-) phenotype (0.029 9%, 22/73 691); 30 cases of the Jk(a-b-) phenotype (0.010 1%, 30/298 250); and 1 case of the k(-) phenotype (0.001 3%, 1/77 382), which was further identified as KELnull phenotype (K0). No p phenotype donors were identified (0/88 528). A total of 228 units of frozen red blood cells were prepared. The screening results were compared and analyzed with rare blood type data from other regions. Conclusion: This study, through a combination of different screening methods, significantly improved the efficiency of rare blood type screening while remaining cost-effective. By conducting large-scale screening and performing data informatization processing, a database and physical repository of rare blood types in the Guangzhou region were successfully established. This provides a strong guarantee for the timely supply of blood to patients with difficult-to-match and rare blood types in the region, effectively enhances the level of transfusion safety in the region, and offers a practical paradigm for constructing a comprehensive blood transfusion support system.
7.Safety and short-term efficacy of single-port robotic transanal total mesorectal excision
Huichao ZHENG ; Weidong TONG ; Bin HUANG ; Qiulin LIAO ; Haijie ZOU ; Feifei HUANG ; Nana WEN ; Jialing LIU ; Fan LI
Chinese Journal of Digestive Surgery 2025;24(6):762-768
Objective:To explore the safety and short-term efficacy of single-port robotic transanal total mesorectal excision (SPr-taTME).Methods:The retrospective and descriptive study was conducted. The clinicopathological data of six patients who underwent SPr-taTME at Daping Hospital of Army Medical University from October to November 2024 were collected. There were 3 males and 3 females, aged (65±5)years. Observation indicators: (1) intraoperative situations; (2) postoperative situations; (3) follow-up. Measurement data with normal distribution were represen-ted as Mean± SD, measurement data with skewed distribution were represented as M(range). Count data were described as absolute numbers. Results:(1) Intraoperative situations. All patients successfully underwent SPr-taTME without conversion to laparotomy or blood transfusion. There was no intraoperative complication such as accidental hemorrhage or adjacent organ injury. No intra-operative adverse events or mortality occurred. The operation time of the 6 patients was 286(range, 240?400)minutes. The time of transanal platform setup and robotic docking was (21±10)minutes, transanal dissection time was (97±45)minutes, and transabdominal dissection time was (90±35)minutes. The volume of intraoperative blood loss was (47±14)mL. Among the six patients, 1 case underwent synchronous transanal and transabdominal surgery, while 5 cases underwent non-synchronous procedures. Specimens were extracted transanally in 5 cases and via an auxiliary abdominal incision in 1 case. The single-port robotic platform was utilized for the abdominal surgery in 3 cases, while laparoscopy was used in 3 cases. Splenic flexure mobilization was performed in 3 cases and omitted in the other 3 cases. Three patients underwent hand-sewn sigmoid colon-anal anastomosis, 1 case underwent modified Bacon pull-through anastomosis, 1 case received stapled sigmoidorectal anastomosis, 1 case underwent sigmoid colostomy without anastomosis due to significant bowel edema. Two cases didn′t undergo intestinal stoma, 2 cases underwent virtual ileostomy, 1 case underwent ileostomy, and 1 case underwent sigmoid colostomy. (2) Postoperative situations. All patients started water drinking and out‐of‐bed activities on postoperative day 1 and liquid diet intake on postoperative day 2. The time to postoperative first flatus was 1(range, 1?3)days, and duration of postoperative hospital stay was (8±2)days.The total number of lymph nodes dissected was 13±2, with the number of positive lymph nodes as 0(range, 0?3) and the distance of distal resection margin as (23±8)mm. Pathological examination of 6 patients showed 1 case in stage T1N0, 2 cases in stage ypT0N0, 1 case in ypT1N0, 1 case in ypT3N1, and 1 case in ypT0N1. The degree of mesorectal integrity was complete in 5 patients and nearly complete in 1 patient. The surgical specimens of 6 patients showed negative in distal, proximal and circumferential margin. (3) Follow-up. All 6 patients completed the 30-day postoperative follow-up. None of the patients experienced postoperative complication such as bleeding, intestinal obstruction or anastomotic leakage. There was no readmission within 30 days after surgery. Digital rectal examination or colonoscopy on postoperative 30 day confirmed no anastomosis-related complications, including stenosis, dehiscence or anastomotic leakage. All 6 patients survived.Conclusion:The SPr-taTME is safe and feasible, with satisfactory short-term efficacy.
8.Application effect of double eyelid plasty with the levator palpebrae superioris aponeurosis fiber flap method
Guohao LI ; Huaiwei LIAO ; Yan SHI ; Yuhan QI ; Huicai WEN
Chinese Journal of Plastic Surgery 2025;41(1):1-7
Objective:To explore the clinical application effect of double eyelid plasty using the levator palpebrae superioris aponeurosis fiber flap.Methods:In March 2021, 3-mm-wide tissue strips were vertically harvested from the mid-upper eyelids from 5 adult Han cadavers with single eyelids, provided by the Anatomy Lab of Nanchang Vocational Health College, and then Masson staining was performed to track the levator aponeurosis fiber. A descriptive case series study was performed. From February 2021 to February 2022, eligible patients who were prospectively collected from the Outpatient Department of the First Affiliated Hospital of Nanchang University underwent double eyelid plasty. The levator palpebrae superioris aponeurosis fiber flap was dissected and repositioned between the orbicularis oculi muscles at the upper and lower edge of incision. The surgical site was closed by a three-layer multi-faceted suture fixation: the muscle of orbicularis oculi at lower edge of incision was sutured to the flap root and tarsal upper edge; the upper and the muscle of orbicularis oculi at the upper and lower edge were sutured to the flap end; the skin was sutured to the muscle of orbicularis oculi and flap end. Postoperatively, incision healing and complications were monitored. Twelve months after surgery, two independent attending plastic surgeons evaluated double eyelid height, curvature, symmetry, and incision scar. The patients were asked to evaluate the effect as satisfied, relatively satisfied, or dissatisfied.Results:Masson staining of 5 lateral upper eyelid specimens revealed that in single-eyelid cases, the levator palpebrae superioris aponeurosis diverged into fibers at the white line, penetrated the orbicularis oculi loosely, and extended to the eyelid margin. Fourty female patients with single or hidden double eyelids were included, aged 18-38 years. After 12 months’ follow-up, all patients had good incision healing without major complications. Twelve months after the operation, the result of doctor evaluation was as followed: 40 cases had proper line height and natural curvature; 38 cases had good line retention, 2 cases had shallower lines; 39 cases had good symmetry, 1 case had asymmetry; 40 cases had no sunken scar. The result of patient satisfaction was as satisfied (35 cases), relatively satisfied (2 cases), dissatisfied (3 cases), with a 92.5% (37/40) satisfaction rate.Conclusion:The levator palpebrae superioris aponeurosis fiber flap method can reconstruct upper eyelid tissue, with natural dynamic double eyelids with few complications and high patient satisfaction.
9.Fish-shaped auricular cartilage framework in Chinese Han women’s comprehensive rhinoplasty
Yiming WEI ; Huicai WEN ; Huaiwei LIAO ; Leqiang XIAO ; Yuhan QI ; Xuenong ZHAN
Chinese Journal of Plastic Surgery 2025;41(8):789-798
Objective:To validate the validity of the auricular cartilage fish-shaped framework in comprehensive rhinoplasty for Chinese Han females.Methods:The clinical data of Chinese Han female patients with short nose and saddle nasal deformity undergoing primary rhinoplasty at the Department of Plastic Surgery, the First Affiliated Hospital of Nanchang University, from August 2021 to October 2023, were retrospectively analyzed. Patients underwent comprehensive rhinoplasty, involving the implantation of an auricular cartilage fish-shaped framework at the nasal tip and a dorsal nasal implant on the nasal bridge.Postoperative follow-up monitored for complications. At 6-month postoperatively, a satisfaction survey was conducted among the patients, their family, or friends using a scale where: very satisfied with improvement=4, moderately satisfied with improvement=3, Slightly satisfied with improvement=2, No improvement=1. Additionally, two plastic surgeons evaluated the postoperative outcomes using a scale where: excellent=5, very good=4, good=3, fair=2, poor=1.Preoperative and 6-month postoperative photographs were processed using Image J software to measure the following parameters: nasolabial angle, nasofacial angle, nasofrontal angle, columellar lobular angle, nasal height, and nasal length. Paired t-tests were used for statistical analysis, with P < 0.05 considered statistically significant. Results:A total of 18 patients were included, with a mean age of (26.4 ± 7.2) years (range: 18-45 years). The mean postoperative follow-up duration was (23.0 ± 9.3) months (range: 6 months to 3 years). All patients recovered well, with no severe complications occurring.The mean patient satisfaction score regarding the postoperative outcome was 3.50 ± 0.62. The mean satisfaction score from family or friends was 3.33 ± 0.69. The two evaluating plastic surgeons rated the outcomes as 4.17 ± 0.85 and 3.94 ± 0.93, respectively, both falling between the excellent and very good categories on the assessment scale.Statistically significant differences (all P < 0.01) were found between preoperative and 6-month postoperative measurements for all assessed nasal parameters. Nasolabial angle: 99.68° ± 2.95° vs. 95.37° ± 1.09°; nasofacial angle: 29.47° ± 2.99° vs. 32.82° ± 2.23°; nasofrontal angle: 140.72° ± 5.18° vs. 133.79° ± 4.43°; columellar lobular angle: 44.86° ± 3.43° vs. 39.23° ± 6.44°; nasal height: (57.52 ± 0.60) mm vs. (58.41 ± 0.39) mm; nasal length: (50.08 ± 0.54) mm vs. (50.64 ± 0.31) mm. Conclusion:The use of the auricular cartilage fish-shaped framework combined with a prosthesis in rhinoplasty, requiring only unilateral or bilateral auricular cartilage, effectively improved low nose and short nose deformities in the majority of Chinese Han females. This validates the favorable outcome of this framework in comprehensive rhinoplasty for this population.
10.Preparation of Monoclonal Antibody to Vasorin in Tree Shrew and Exploration of Its Application
Meizhen OU ; Yongfeng LI ; Sha WEN ; Zhouxiang LIAO ; Xuejing HUANG ; Lichao YANG ; Min HE
Laboratory Animal and Comparative Medicine 2025;45(5):611-622
Objective To obtain tree shrew Vasorin(VASN)recombinant protein through prokaryotic expression and purification,prepare monoclonal antibody against tree shrew VASN by immunizing mice with this protein,and preliminarily evaluate its application value.Methods Reverse transcription-polymerase chain reaction(RT-PCR)was used to amplify the full-length sequence of tree shrew VASN gene in vitro.The tree shrew VASN gene fragment was inserted into pET-30a vector to construct pET-30a-VASN recombinant plasmid.The recombinant plasmid was subjected to double digestion with BamH Ⅰ and Sal Ⅰfor identification,and its correctness was further verified by sequencing.The recombinant plasmid with correct sequencing was transformed into BL21(DE3)competent cells,and isopropyl β-D-thiogalactoside(IPTG)was used to induce expression of VASN recombinant protein.Proteins were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE),and the VASN recombinant protein was purified by KCI.Purified recombinant protein was used to immunize BALB/c mice for four times,and serum antibody titer was detected by enzyme-linked immunosorbent assay(ELISA).Splenocytes from mice with serum antibody titer above 1:10 000 were used for cell fusion with myeloma cells.Hypoxanthine-aminopterin-thymidine(HAT)culture medium was first used to screen hybridoma cells.ELISA was used to screen positive hybridoma cell lines that could secrete specific antibodies,and monoclonal hybridoma cell lines were obtained by limiting dilution method.VASN monoclonal antibodies were prepared in large quantities by ascites induction method,purified using rProtein G,and the affinity and in vitro reaction specificity of the monoclonal antibodies were detected by ELISA and Western blotting.Results The full-length sequence of the tree shrew VASN gene was successfully amplified and the recombinant plasmid vector of tree shrew pET-30a-VASN was constructed.The sequence obtained by sequencing of the recombinant plasmid vector was identical to the tree shrew VASN target gene sequence.Recombinant protein VASN mainly existed in the form of inclusion bodies,and the purity after purification reached 90%,meeting the requirements of subsequent immunization experiments.After four immunizations with recombinant protein VASN,mouse serum antibody titer reached 1:729 000.Monoclonal positive hybridoma cell lines were obtained through ascites induction and purification,and the constant affinity value of monoclonal antibodies measured by ELISA reached 2.59x107 L/mol.Western blotting results showed that the tree shrew VASN monoclonal antibody could bind to tree shrew VASN recombinant protein,but it showed no binding reaction with porcine retinol-binding protein 4 recombinant protein,human VASN-leucine rich repeat recombinant protein,or bovine serum albumin.Anti-tree shrew VASN monoclonal antibody could specifically recognize VASN protein in tree shrew heart,liver,spleen,lung,kidney and muscle,with clear bands and clean background.Immunohistochemical detection results showed that this monoclonal antibody could recognize VASN protein in tree shrew spleen,lung,and tree shrew immortalized fibroblasts with high VASN mRNA expression levels,and the detection results were positive.Conclusion Monoclonal antibody against tree shrew VASN is successfully prepared.This antibody can be used for immunohistochemical detection of tree shrew immortalized fibroblasts,spleen tissue,and lung tissue,providing an important tool for further research on the function of VASN in tree shrew models.

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