1.Study of DNA Transfer from Hand Exfoliated Cells with Non-permeable Carriers
Li YUAN ; Kainan ZOU ; Shicheng HAO ; Haixiao DENG ; Jinpei ZHANG ; Libin WU ; Hang HE
Journal of Sun Yat-sen University(Medical Sciences) 2024;45(5):818-825
[Objective]To investigate primary and secondary transfer of exfoliated cells from human hands on non-porous substrates such as plastic steering wheel or computer mouse.[Methods]DNA detection sensitivity and detection limit for mixed DNA profiling were examined to understand our laboratory's ability to test for trace DNA.Forensic swabs were used to collect samples from volunteers'one-hour-long unwashed hands,substrates touched by volunteers'immediately or 30 min following shaking hands,and individual A's daily-use substrates touched by individual B and then by individual A again.Simulations were conducted to assess the potential for introduction of another person's exfoliated cells from hands into routine casework samples.[Results]Our laboratory can obtain a full DNA profile from as little as 0.020 ng of DNA and detect minor components in a 1:9 mixed DNA sample.85%of samples from unwashed hands yielded a full DNA profile.Primary transfer of a full DNA profile was found in 77%of substrates touched by volunteers'dominant hand 30 min after hand washing,allowing differentiation between good and poor shedders,with no significant difference in genders and substrate types.75%of substrates touched 30 min after hand washing and then immediately following handshaking yielded the other individual's DNA profile(secondary transfer),with the number of short tandem repeat(STR)loci detected ranging from 0 to 23;the percentage and number decreased substantially when the substrates were touched 30 minutes later.No foreign DNA was detected in routine casework samples with introduced exfoliated cells from hands.When two individuals took turns touching items with their hands,the major contributor to the DNA profile was not always the individual who made the last contact.[Conclusions]Primary and secondary DNA transfer can be detected on non-porous substrates,and based on the deposit of hand exfoliated cells,individuals can be categorized as good or poor shedders,which is an important factor affecting detection of DNA transfer.Besides considering the laboratory's DNA detection sensitivity,if DNA is detected on substrates by hand contact,we need to take into account the potential for secondary transfer at different levels of activity when interpreting the results.
2.Analysis on effects of EFTR-C and ESE for treating≤1.5 cm intraluminal growth type gastric stromal tumor
Bin HUANG ; Haixiao FU ; Bin LI ; Xuhua XIAO
Chongqing Medicine 2024;53(17):2583-2587
Objective To compare the effectiveness and safety of transparency cap-assisted endoscopic full-thickness resection(EFTR-C)and endoscopic submucosal excavation(ESE)in the treatment of ≤1.5 cm intracavity growth type gastric stromal tumor.Methods A retrospective analysis was performed on the medi-cal records of 63 patients with ≤1.5 cm intraluminal growth type gastric stromal tumor treated in this hospi-tal from June 2019 to September 2023.The patients were divided into the EFTR-C group(n=33)and ESE group(n=30)according to surgical methods.The general data,operation situation,postoperative situation and follow-up were compared between the two groups.Results There was no statistically significant differ-ence in the intraoperative pneumoperitoneum rate,purse string suture rate,monoblock resection rate,complete resection rate and complication occurrence rate between the two groups(P>0.05).The operative time,inci-dence rate of intraoperative bleeding,number of titanium clips,postoperative fasting time,postoperative hospi-talization duration and operation costs had statistical difference between the two groups(P<0.05).Postoper-ative wound healing was good in both groups without recurrence,metastasis and death.Conclusion EFTR-C for the treatment of ≤1.5 cm intracavity growth type gastric stromal tumor has the advantages of simple op-eration,short operation time,low costs and rapid recovery compared with ESE.
3.Screening,validation,and functional analysis of Brucella secretory BspE interac-ting host proteins
Shuanghong YIN ; Xiaoyu DENG ; Hongyan LIU ; Haixiao WANG ; Caixia YI ; Yincui LI ; Xin SUN ; Shuli WANG ; Jihai YI ; Junbo ZHANG
Chinese Journal of Veterinary Science 2024;44(7):1438-1447,1457
In order to explore the role of BspE protein in Brucella infection,yeast two-hybrid tech-nique was used to screen host cell proteins that interact with BspE protein.The constructed BspE recombinant plasmid pGBKT7-BspE was used as bait plasmid to hybridize with the RAW264.7-cD-NA library of mouse mononuclear macrophages by yeast two-hybridization technique.The positive clones were extracted by plasmid,sequenced and co-immunoprecipitation to determine the host cell proteins that could interact with BspE.The subcellular localization of BspE proteins was analyzed by confocal laser microscopy.The physical and chemical properties,protein structure and function of BspE interacting proteins were analyzed by bioinformatics.The siRNA for one of the BspE inter-acting proteins was synthesized,the expression of its gene was silenced in HEK293T cells,and the silenced cells was infected with Brucella M5-90 and the number of intracellular bacteria was coun-ted.The results showed that the decoy plasmid pGBKT7-BspE was successfully constructed,and the plasmid could express BspE protein in yeast.Eight positive clones were obtained from the host cell genome library by yeast two-hybridization.The positive clones were identified as RBM27 and PCBP1 by sequencing,backcross and co-immunoprecipitation.Bioinformatics was used to predict the cell location,protein structure and amino acid composition of RBM27 and PCBP1.After siRNA interference,the expression level of PCBP1 was significantly decreased and the amount of M5-90 in the cell was increased.Brucellosis secreted protein BspE interacts with host proteins RBM27 and PCBPl,and PCBP1 negatively regulates the proliferation of Brucellosis.
4.Exosomes derived from gastric cancer cells trigger M2 polarization of hepatic Kupffer cells via miRNA to promote the formation of pre-metastatic hepatic niche
Xuan ZHANG ; Wei LIU ; Haixiao FU ; Tengteng LI ; Hao LIU ; Wei FU ; Kai WANG
Chinese Journal of Microbiology and Immunology 2024;44(9):762-770
Objective:To investigate the influence of the interaction between gastric cancer (GC) cell-derived exosomes and hepatic Kupffer cells on GC with liver metastasis and analyze the potential mechanism.Methods:Cells with high hepatic metastatic potential (MKN 45-HL) were constructed from a parental GC cell line (MKN 45) using a nude mouse model and methods of viral transfection and flow sorting. Exosomes were collected using ultra-centrifugation and characterized by electron microscopy, nanoparticle tracking system and Western blot. A nude mouse model of liver metastasis induced by GC cell-derived exosomes was constructed, and the development of liver metastases was monitored by live imaging. The regulatory effects of GC cell-derived exosomes on macrophage polarization were assessed by cell culture, qRT-PCR, and immunofluorescence staining. Using the omics analysis of exosomal miRNA and qRT-PCR, the molecular targets by which exosomes specifically promoting macrophage M2 polarization were screened and validated.Results:GC cell-derived exosomes were mainly concentrated in the liver, most of which were ingested by intrahepatic macrophages, and could promote macrophages to M2 polarization in both in vitro culture and nude mice. Both groups of mice trained with MKN 45 and MKN 45-HL exosomes showed obvious liver metastases after mouse forestomach carcinoma (MFC) cells injection through the spleen, and MKN 45-HL exosomes showed a much stronger ability to promote hepatic macrophage M2 polarization and liver metastasis of MFC cells. Moreover, the miRNA omics analysis revealed a lot of differentially expressed miRNAs between MKN 45-derived and MKN 45-HL-derived exosomes. The expression of miR-519a-3p increased significantly in the exosomes derived from MKN 45-HL cell line and the clinical serum of GC patients with liver metastasis. It was found that miR-519a-3p could be internalized by macrophages through exosomes delivery. Furthermore, the miR-519a-3p in exosomes from patient′s serum had a predictive value for GC with liver metastasis and was closely associated with the prognosis of GC patients with liver metastasis. Conclusions:GC cell-derived exosomes trigger M2-like polarization of hepatic Kupffer cells via miR-519a-3p, thus promoting the progression of liver metastasis in GC and playing a critical role in shaping the pre-metastatic liver niche in gastric cancer. This study provides a new perspective on the mechanism of GC with liver metastasis and reveal potential targets for future therapeutic strategies.
5.Clinical efficacy of da Vinci Xi surgical system assisted programmed six-hole method anterior resection of rectal cancer
Kai WANG ; Wei FU ; Haixiao FU ; Tengteng LI ; Hao LIU ; Jingjing HE ; Jun SONG ; Xuan ZHANG
Chinese Journal of Digestive Surgery 2023;22(6):769-778
Objective:To investigate the clinical efficacy of da Vinci Xi surgical system assisted programmed six-hole method anterior resection of rectal cancer.Methods:The retrospec-tive cohort study was conducted. The clinicopathological data of 102 patients with middle and low rectal cancer who were admitted to the Affiliated Hospital of Xuzhou Medical University from August 2020 to June 2021 were collected. There were 62 males and 40 females, aged (53±12)years. Of the 102 patients, 51 cases undergoing da Vinci Xi surgical system assisted programmed six-hole method anterior resection of rectal cancer were divided into the robotic group and 51 cases undergoing laparoscopic anterior resection of rectal cancer were divided into the laparoscopic group. Observa-tion indicators: (1) treatment; (2) postoperative pathological examination; (3) follow-up. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was conducted using the independent sample t test. Measurement data with skewed distribution were represented as M(range), and comparison between groups was conducted using the Mann-Whitney U test. Count data were described as absolute numbers, and comparison between groups was conducted using the chi-square test or Fisher exact probability. Repeated measurement data were analyzed using the repeated ANOVA. Results:(1) Treatment. All patients of the two groups under-went radical resection of rectal cancer successfully, and none of patient with intraoperative blood transfusion, conversion to open surgery, and death within 30 days after surgery. The operation time, volume of intraoperative blood loss, number of lymph nodes dissected, time to postoperative first flatus, time to postoperative first liquid food intake, time to postoperative catheter removal, cases with postoperative pain grading as grade 1, grade 2, grade 3, grade 4, cost of treatment were (170±12)minutes, (73±50)mL, 23±6, (35.1±9.4)hours, (2.1±0.8)days, (2.9±2.7)days, 13, 15, 17, 6, (7.1±4.5) ten thousand yuan in patients of the robotic group, versus (153±22)minutes, (119±66) mL, 15±4, (40.7±1.9)hours, (2.9±0.4)days, (5.3±2.1)days, 6, 7, 26, 12, (6.7±1.6) ten thousand yuan in patients of the laparoscopic group, showing significant differences in the above indicators between the two groups ( t=6.79, -4.46,20.09, -3.01, -5.54, -16.69, Z=-2.87, t=4.22, P<0.05). (2) Postoperative patho-logical examination. The tumor diameter, length of specimen resected, distance of upper resection margin to tumor, distance of lower resection margin to tumor, cases with mesorectal specimens as integrity and mostly integrity, cases with tumor differentiation as high differentiation, moderate differentiation, low differentiation, cases with postoperative TNM staging as stage Ⅰ, stage Ⅱ, stage Ⅲ were (3.8±1.1)cm, (18.7±3.2)cm, (11.8±3.6)cm, (2.7±0.8)cm, 48, 3, 4, 41, 6, 6, 17, 28 in patients of the robotic group, versus (3.7±1.0)cm, (18.3±2.8)cm, (10.2±2.7)cm, (2.5±0.6)cm, 46, 5, 6, 39, 6, 5,20, 26 in patients of the laparoscopic group, showing no significant difference in the above indicators between the two groups ( t=1.72, 1.29, 1.64, 1.11, χ2=0.14, Z=-0.42, -0.26, P>0.05). Cases with positive circumferential margin and cases with destruction of mesentery was 0 and 0 in patients of the robotic group, versus 1 and 1 in patients of the laparoscopic group, showing no significant difference in the above indicators between the two groups ( P>0.05). (3) Follow-up. All patients in the two groups were followed up for 12 months after surgery and none of patient had postoperative local recurrence and distant metastasis of tumors. The anal incontinence score, low anterior resection syndrome score, international prostate symptom score, night urination score, international index of erectile score, female sexual function index score in patients of the robotic group were 0, 12.25±1.08, 4.43±0.33, 0.49±0.09, 24.07±2.75, 65.84±1.79 before surgery and 1.34±0.11, 18.11±3.54, 4.03±0.26, 1.08±0.28, 22.63±2.03, 38.57±6.13 at postoperative 12 months, respectively. The above indicators in patients of the laparoscopic group were 0, 12.60±1.11, 4.56±0.36, 0.46±0.07, 23.11±2.77, 66.31±1.73 before surgery and 1.99±1.33,20.85±6.19, 6.43±1.78, 2.27±0.23, 21.00±2.73, 27.62±8.20 at postoperative 12 months, respectively. There were significant differences in the above indicators between the two groups ( P<0.05). Conclusions:The oncological effects of da Vinci Xi surgical system assisted programmed six-hole method anterior resection of rectal cancer and lapa-roscopic anterior resection of rectal cancer are comparable. However, robotic surgery is superior to laparoscopic surgery in terms of intraoperative bleeding, lymph node dissection, gastrointestinal function recovery, and pelvic autonomic nerve protection.
6.Safety of the strategy of minimizing intestinal resection during surgery for pelvic radiation- induced terminal small intestinal stenosis
Kai WANG ; Xiaodong NI ; Bangjian BIAN ; Xuan ZHANG ; Haixiao FU ; Tengteng LI ; Hao LIU ; Wei FU ; Jun SONG ; Jian WANG
Chinese Journal of Gastrointestinal Surgery 2023;26(10):947-954
Objective:To investigate the efficacy of strategies for minimizing small bowel resection during surgery for pelvic radiation-induced terminal small intestinal stenosis in preventing postoperative complications such as anastomotic leakage and short bowel syndrome.Methods:This was a retrospective cohort study. There are two subtypes of chronic radiation enteritis (CRE) with combined intestinal stenosis and intestinal obstruction: (1) Type I: terminal ileal lesions with a normal ileal segment of 2–20 cm between the ileal lesion and ileocecal junction; and (2) Type II: the lesion is located in the small bowel at a distance from the ileocecal region, usually accompanied by extensive damage to the bowel segments outside the lesion. The indications for minimal bowel resection are as follows: (1) diagnosis of Type I small bowel CRE; (2) absence of radiological evidence of rectosigmoid damage; and (3) absence of colonic obstruction. The contraindications are: (1) stenotic, penetrating lesions of the distal cecum; (2) emergency surgery; (3) recurrence of malignant tumor or history of radiotherapy for recurrent malignant tumor; (4) interval between radiotherapy and surgery <6 months; and (5) history of preoperative small bowel resection or abdominal chemotherapy. Case data of 40 patients with Type I CRE who met the above criteria and had undergone minimal bowel resection between April 2017 and December 2019 were retrospectively analyzed (minimal bowel resection group; including 13 patients from Jinling Hospital, 16 from the Ninth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, and 11 from the Affiliated Hospital of Xuzhou Medical University). Forty patients with Type I CRE who had undergone resection of intestinal stenosis lesions and the ileocecal region between October 2015 and March 2017 were included as historical controls (conventional resection group; all from Jinling Hospital). The specific strategy for minimal bowel resection was one-stage partial ileal resection+ileo anastomosis+protective small bowel stoma. In contrast, conventional resection comprised ileocecal resection+ileocecal-ascending colon anastomosis. Postoperative complications, intraoperative and postoperative recovery, and changes in postoperative quality of life were analyzed in both groups. The severity of postoperative complications was assessed by Clavien-Dindo and the Comprehensive Complication Index (CCI). Karnofsky performance scores (KPS) were used to evaluate the quality of life of patients in the two groups preoperatively and postoperatively. The higher the KPS score, the better the quality of life.Results:Baseline patient characteristics did not differ significantly between the two groups ( P>0.05). Compared with the conventional resection group, the length of small bowel resected in the minimal bowel resection group (51 [20–200] cm vs. 91 [60–200] cm, Z=5.653, P<0.001), duration of postoperative total enteral nutrition [9 (3–18) days vs. 12 (4–50) days, Z=2.172, P=0.030], and duration of postoperative hospital stay [17 (9–24) days vs 29 (13–57) days, Z=6.424, P<0.001] were shorter; all of these differences are statistically significant. The overall incidence of postoperative complications was lower in the minimal bowel resection group than in the conventional resection group [20.0% (8/40) vs. 70.0% (28/40), χ 2=19.967, P<0.001], These comprised short bowel syndrome [5.0% (2/40) vs. 25.0% (10/40), χ 2=6.274, P=0.012], anastomotic leakage or fistula [2.5% (1/40) vs. 22.5% (9/40), χ 2=7.314, P=0.014], and pleural effusion [7.5% (3/40) vs. 25.0% (10/40), χ 2=4.500, P=0.034], all of which occurred less often in the minimal bowel resection than conventional resection group. The CCI index was also lower in the minimal bowel resection group than in the conventional resection group [CCI>40: 2.5% (1/40) vs. 12.5% (5/40), Z=18.451, P<0.001]. KPS scores were higher in the minimal bowel resection group 1 and 3 months postoperatively than they had been 1 day preoperatively (79.9±4.7 vs. 75.3±4.1, 86.2±4.8 vs. 75.3±4.1, both P<0.05). In the minimal bowel resection group, seven patients were satisfied with their current quality of life and refused to undergo stoma reduction at follow-up and one deferred stoma reduction because of rectal bleeding. The remaining 32 patients underwent stoma reduction 3 to 12 months after surgery, 26 of whom underwent ileo-cecal anastomosis. The remaining six underwent resection of the stoma and anastomosis of the ileum to the ascending colon. Conclusions:The strategy of minimal small bowel resection in patients with radiation-induced bowel injuries reduces the length of resected small bowel, decreases the risk and severity of postoperative complications, and is associated with a better prognosis and quality of life than conventional resection.
7.Safety of the strategy of minimizing intestinal resection during surgery for pelvic radiation- induced terminal small intestinal stenosis
Kai WANG ; Xiaodong NI ; Bangjian BIAN ; Xuan ZHANG ; Haixiao FU ; Tengteng LI ; Hao LIU ; Wei FU ; Jun SONG ; Jian WANG
Chinese Journal of Gastrointestinal Surgery 2023;26(10):947-954
Objective:To investigate the efficacy of strategies for minimizing small bowel resection during surgery for pelvic radiation-induced terminal small intestinal stenosis in preventing postoperative complications such as anastomotic leakage and short bowel syndrome.Methods:This was a retrospective cohort study. There are two subtypes of chronic radiation enteritis (CRE) with combined intestinal stenosis and intestinal obstruction: (1) Type I: terminal ileal lesions with a normal ileal segment of 2–20 cm between the ileal lesion and ileocecal junction; and (2) Type II: the lesion is located in the small bowel at a distance from the ileocecal region, usually accompanied by extensive damage to the bowel segments outside the lesion. The indications for minimal bowel resection are as follows: (1) diagnosis of Type I small bowel CRE; (2) absence of radiological evidence of rectosigmoid damage; and (3) absence of colonic obstruction. The contraindications are: (1) stenotic, penetrating lesions of the distal cecum; (2) emergency surgery; (3) recurrence of malignant tumor or history of radiotherapy for recurrent malignant tumor; (4) interval between radiotherapy and surgery <6 months; and (5) history of preoperative small bowel resection or abdominal chemotherapy. Case data of 40 patients with Type I CRE who met the above criteria and had undergone minimal bowel resection between April 2017 and December 2019 were retrospectively analyzed (minimal bowel resection group; including 13 patients from Jinling Hospital, 16 from the Ninth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, and 11 from the Affiliated Hospital of Xuzhou Medical University). Forty patients with Type I CRE who had undergone resection of intestinal stenosis lesions and the ileocecal region between October 2015 and March 2017 were included as historical controls (conventional resection group; all from Jinling Hospital). The specific strategy for minimal bowel resection was one-stage partial ileal resection+ileo anastomosis+protective small bowel stoma. In contrast, conventional resection comprised ileocecal resection+ileocecal-ascending colon anastomosis. Postoperative complications, intraoperative and postoperative recovery, and changes in postoperative quality of life were analyzed in both groups. The severity of postoperative complications was assessed by Clavien-Dindo and the Comprehensive Complication Index (CCI). Karnofsky performance scores (KPS) were used to evaluate the quality of life of patients in the two groups preoperatively and postoperatively. The higher the KPS score, the better the quality of life.Results:Baseline patient characteristics did not differ significantly between the two groups ( P>0.05). Compared with the conventional resection group, the length of small bowel resected in the minimal bowel resection group (51 [20–200] cm vs. 91 [60–200] cm, Z=5.653, P<0.001), duration of postoperative total enteral nutrition [9 (3–18) days vs. 12 (4–50) days, Z=2.172, P=0.030], and duration of postoperative hospital stay [17 (9–24) days vs 29 (13–57) days, Z=6.424, P<0.001] were shorter; all of these differences are statistically significant. The overall incidence of postoperative complications was lower in the minimal bowel resection group than in the conventional resection group [20.0% (8/40) vs. 70.0% (28/40), χ 2=19.967, P<0.001], These comprised short bowel syndrome [5.0% (2/40) vs. 25.0% (10/40), χ 2=6.274, P=0.012], anastomotic leakage or fistula [2.5% (1/40) vs. 22.5% (9/40), χ 2=7.314, P=0.014], and pleural effusion [7.5% (3/40) vs. 25.0% (10/40), χ 2=4.500, P=0.034], all of which occurred less often in the minimal bowel resection than conventional resection group. The CCI index was also lower in the minimal bowel resection group than in the conventional resection group [CCI>40: 2.5% (1/40) vs. 12.5% (5/40), Z=18.451, P<0.001]. KPS scores were higher in the minimal bowel resection group 1 and 3 months postoperatively than they had been 1 day preoperatively (79.9±4.7 vs. 75.3±4.1, 86.2±4.8 vs. 75.3±4.1, both P<0.05). In the minimal bowel resection group, seven patients were satisfied with their current quality of life and refused to undergo stoma reduction at follow-up and one deferred stoma reduction because of rectal bleeding. The remaining 32 patients underwent stoma reduction 3 to 12 months after surgery, 26 of whom underwent ileo-cecal anastomosis. The remaining six underwent resection of the stoma and anastomosis of the ileum to the ascending colon. Conclusions:The strategy of minimal small bowel resection in patients with radiation-induced bowel injuries reduces the length of resected small bowel, decreases the risk and severity of postoperative complications, and is associated with a better prognosis and quality of life than conventional resection.
8.Application of scenario simulation teaching combined with Mini-CEX in standardized residency training of general surgery
Xuan ZHANG ; Haixiao FU ; Hao LIU ; Kai WANG ; Tengteng LI ; Wei FU
Chinese Journal of Medical Education Research 2022;21(1):81-84
Objective:To study the application effect of scenario simulation teaching combined with mini-clinical evaluation exercise (Mini-CEX) in the standardized residency training of general surgery.Methods:The study included in 62 trainees who had standardized residency training in the Department of General Surgery of the Affiliated Hospital of Xuzhou Medical University From July 2019 to July 2020. The subjects were randomly divided into traditional teaching group (control group) and scenario simulation teaching combined with Mini-CEX teaching group (experimental group), with 31 students in each group. The scores of the entrance examination, Mini-CEX scores and the evaluation of teaching effect were compared between the two groups. SPSS 21.0 was used to perform t test on the test scores, Mini-CEX scores and teaching effective evaluation scores of the two groups. Results:①The theoretical scores of the experimental group [(82.48 ± 6.02) points] were significantly higher than those of the control group [(77.32±6.25) points], with significant differences ( t=3.31, P<0.01). The clinical practice scores of the experimental group [(88.96 ± 2.93) points] were significantly higher than those of the control group [(80.87±5.41) points], with significant differences ( t=7.33, P<0.01). ②Mini-CEX scores of the experimental group were higher than those of the control group ( P<0.01). ③Through the teaching questionnaire, the scores of the experimental group were higher than those of the control group ( P<0.01). Conclusion:Scenario simulation teaching combined with Mini-CEX has achieved good results in the standardized residency training of general surgery, which could be used as a new clinical teaching mode.
9.Analysis of genetic variant in a Chinese pedigree with hereditary factor XIII deficiency
Haixiao XIE ; Lihong YANG ; Huinan XIA ; Yanhui JIN ; Xiaolong LI ; Shuting JIANG ; Yaoyao XU ; Mingshan WANG
Chinese Journal of Medical Genetics 2022;39(7):708-712
Objective:To explore the genetic basis for a patient with factor XIII (FXIII) deficiency.Methods:All exons of the F13A1 and F13B genes were amplified by PCR and sequenced directly. The sequencing was performed with a reverse primer if a variant was found. Conservation of variant site was analyzed by the ClustalX software. Four online bioinformatic software including MutationTaster, PolyPhen-2, PROVEAN and SIFT were used to predict the function of the mutation site. The Swiss-PdbViewer software was applied to analyze the changes in the protein model and intermolecular force. Results:The proband was found to harbor a novel c. 515G>C (p.Arg171Pro) variant of the F13A1 gene. The corresponding amino acid Arg171 is conserved among homologous species. Bioinformatic analysis indicated that Arg171Pro variant may affect the protein function. Protein model analysis showed that in the wild-type, there is one hydrogen bond between Arg171 and Pro27; one hydrogen bond between Arg171 and Thr28; two hydrogen bonds between Arg171 and Glu102. When Arg171 was mutated to Pro171, the three hydrogen bonds between Arg171 and Pro27, Glu102 are all disappeared and formed a new benzene ring which might affect the stability of the protein structure. No variant was found in the F13B gene. Conclusion:The Arg171Pro variant may account for the decreased FXIII level. Above finding has enriched the spectrum of F13A1 gene variants.
10. Antagonism of Protease-Activated Receptor 4 Protects Against Traumatic Brain Injury by Suppressing Neuroinflammation via Inhibition of Tab2/NF-κB Signaling
Jianing LUO ; Xun WU ; Haixiao LIU ; Wenxing CUI ; Wei GUO ; Kang GUO ; Hao GUO ; Kai TAO ; Fei LI ; Yingwu SHI ; Dayun FENG ; Guodong GAO ; Yan QU ; Hao YAN
Neuroscience Bulletin 2021;37(2):242-254
Traumatic brain injury (TBI) triggers the activation of the endogenous coagulation mechanism, and a large amount of thrombin is released to curb uncontrollable bleeding through thrombin receptors, also known as protease-activated receptors (PARs). However, thrombin is one of the most critical factors in secondary brain injury. Thus, the PARs may be effective targets against hemorrhagic brain injury. Since the PAR1 antagonist has an increased bleeding risk in clinical practice, PAR4 blockade has been suggested as a more promising treatment. Here, we explored the expression pattern of PAR4 in the brain of mice after TBI, and explored the effect and possible mechanism of BMS-986120 (BMS), a novel selective and reversible PAR4 antagonist on secondary brain injury. Treatment with BMS protected against TBI in mice. mRNA-seq analysis, Western blot, and qRT-PCR verification in vitro showed that BMS significantly inhibited thrombin-induced inflammation in astrocytes, and suggested that the Tab2/ERK/NF-κB signaling pathway plays a key role in this process. Our findings provide reliable evidence that blocking PAR4 is a safe and effective intervention for TBI, and suggest that BMS has a potential clinical application in the management of TBI.

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