1.Baculovirus expression system-based expression of horseshoe crab factor C and its activity.
Lan LAN ; Huanlei LIU ; Hao NAN ; Sijun HE ; Wangcheng SONG ; Yunlong WANG ; Xinjuan FAN ; Xiangbo WAN ; Xiaodong XU
Chinese Journal of Biotechnology 2025;41(4):1428-1439
Endotoxins are common exogenous pyrogens. Excessive endotoxins in medical devices and injections can lead to serious consequences such as sepsis, septic shock, and even death. Therefore, endotoxin detection plays a crucial role in medical, pharmaceutical, and food sectors. The wide application of Limulus amebocyte lysate (LAL) has led to a sharp decline in the number of horseshoe crabs. Moreover, the LAL assay has limitations such as interbatch variations and difficulty in quantification. The recombinant factor C (rFC) assay is stable between batches, highly sensitive, and capable of quantitation, and thus it can be used as an alternative for the LAL assay. However, the high cost and complex procedures involved in producing recombinant factor C have limited the widespread application of this method. In order to simplify the preparation and reduce the production cost of recombinant factor C, this study focuses on the production of recombinant factor C based on the baculovirus expression system. Multiple measures such as a high-yield and anti-apoptotic vector qBac-IIIG, the optimal signal peptide, and the optimized codon were used to reach the goal of endotoxin detection with cell supernatant. This method simplifies the steps of protein purification. The sensitivity of the supernatant reached 0.05 EU/mL in a 1-L fermentation system, and 500 000 detecting reactions can be supported per liter of fermentation broth. This study increases the yield and activity of recombinant factor C, simplifies the procedures of protein purification, and reduces the cost, laying a foundation for the promotion and application of recombinant factor C in endotoxin detection.
Animals
;
Recombinant Proteins/genetics*
;
Horseshoe Crabs/chemistry*
;
Baculoviridae/metabolism*
;
Endotoxins/analysis*
;
Protein C/biosynthesis*
;
Genetic Vectors/genetics*
;
Arthropod Proteins/genetics*
;
Enzyme Precursors
;
Serine Endopeptidases
2.Comparison of the clinical efficacy between biopsy forceps polypectomy with submucosal injection and traditional biopsy forceps polypectomy for diminutive colorectal polyps
Qiuli WU ; Yuanli LI ; Chenwei ZHENG ; Xiangbo CHEN ; Qinwei XU
Chinese Journal of Digestive Endoscopy 2024;41(12):979-984
Objective:To explore the clinical efficacy of biopsy forceps polypectomy with submucosal injection for diminutive colorectal polyps.Methods:The patients with diminutive colorectal polyps (long diameter≤5 mm in size) who received polypectomy in Quanzhou First Hospital Affiliated to Fujian Medical University from May 2021 to May 2022 were prospectively recruited and divided into the observation group (biopsy forceps polypectomy with submucosal injection) and the control group (traditional biopsy forceps polypectomy). The location, morphology, long diameter, the complete polypectomy rate under direct vision, the time of resection and endoscopy withdrawal, pathological results, specimen collection rate, the incidence of complications and endoscopic treatment cost were analyzed. The clinical efficacy of the two methods were compared.Results:A total of 292 patients were included in the study, with 146 in the observation group and 146 in the control group. There were 258 polyps in the observation group and 252 polyps in the control group. The complete polypectomy rate was higher in the observation group than that in the control group [100.0% (258/258) VS 90.1% (227/252), χ2=26.915, P=0.001] with clear vision. The incidence of complications in the observation group was lower than that in the control group (1/146 VS 13/146, χ2=10.804, P=0.001) , along with the lower treatment cost (173.7±15.9 yuan VS 184.0±53.8 yuan, Z=-2.777, P=0.005). The resection time in the observation group was longer than that in the control group, but the difference was not statistically significant [10 (9,11) min VS 9 (9,11) min, Z=-0.528, P=0.597]. There was no significant difference in the polyp position, long diameter, morphology, pathological classification, or the specimen collection rate between the two groups ( P>0.05). Conclusion:Biopsy forceps polypectomy with submucosal injection is safe for diminutive colorectal polyps, and it demonstrates a higher complete polypectomy rate, lower complication rates, and reduced treatment costs compared with traditional biopsy forceps polypectomy. It is a new perspective for managing diminutive colorectal polyps.
3.Eosinophil infiltration in cervical lesion and cervical cancer tissues and their clinical significances
Yanyan LU ; Xiangbo XU ; Yamei WU ; Yuqi LIU ; Han WANG ; Lijuan YANG ; Zhenjiang WANG ; Zishen XIAO ; Yanbo LIU
Journal of Jilin University(Medicine Edition) 2024;50(6):1691-1702
Objective:To discuss the differences in eosinophil(EOS)infiltration in cervical tissue and its relationship with cervical-related diseases,and to clarify the effect of EOS on the occurrence and development of cervical intraepithelial neoplasia(CIN)and cervical cancer.Methods:The clinical data of 256 patients with cervical diseases were collected and divided into cervical cancer group(n=46,including 26 cases of squamous cell carcinoma,15 cases of adenocarcinoma,and 5 cases of adenosquamous carcinoma),chronic cervicitis group(n=50),CIN stage Ⅰ group(n=50),CIN stage Ⅱ group(n=50),CIN stage Ⅲ group(n=30),and normal group(adjacent normal cervical tissue,n=30)based on their conditions.Colposcopy was used to observe the morphology of cervical tissue of the patients in various groups;thin-layer liquid-based cytology test(TCT)was used to observe the morphology of the cervical exfoliated cells in various groups;hybrid capture-chemiluminescence method was used to detect the human papillomavirus(HPV)infection in cervical tissue of the patients in various groups;HE staining was used to observe the pathomorphology of cervical tissue of the patients in various groups;Congo red staining was used to detect the numbers of EOS infiltration in cervical tissue of the patients in various groups;Pearson correlation analysis was used to analyze the correlation between the number of EOS infiltration and the malignancy degree of cervical cancer.Results:The cervical surface of the patients in normal group was smooth and pink,with uniformly distributed capillaries;the cervical surface of the patients in chronic cervicitis group showed red inflammatory changes,with some accompanied by Nabothian cysts and varying degrees of erosion and ulcers;the patients in CIN stage Ⅰ,CIN stage Ⅱ,and CIN stage Ⅲ groups showed epithelial ulcers,thickening,and irregular morphology,with mosaic and punctate vessels;the cervical surface of the patients in cervical cancer group showed raised areas with neoplasms and necrotic ulcers,and they were fragile and prone to bleeding.After acetic acid staining,no obvious changes of the patients in normal group were observed.The cervix of the patients in chronic cervicitis group showed slight white changes that lasted for a short time;in CIN stage Ⅰ,CIN stage Ⅱ,and CIN stage Ⅲ groups,irregular thin acetowhite epithelium with map-like borders was observed,with increasingly acetowhite reactions and larger areas as the stages advanced.The cervix of the patients in cervical cancer group showed thick acetowhite epithelium that lasted longer,with rigid and clear contours.After iodine staining,the cervix of the patients in normal group was brown,with uniform coloration;the cervix of the patients in chronic cervicitis group showed poor coloration in inflammatory lesion areas;the cervix of the patients in CIN stage Ⅰ group showed iodine coloration in metaplastic areas,while the cervix of the patients in CIN stage Ⅲ group showed poor coloration in larger lesion areas;the cervix of the patients in cervical cancer group showed irregular surfaces with cauliflower-like growth and no coloration after iodine staining,appearing orange-yellow or mustard yellow.The TCT observation results showed there were no heteromorphic cells and few inflammatory cells in cervical exfoliated cells of the patients in infiltration in normal group;there were numerous neutrophils and EOS in exfoliated cervical cells without heteromorphic cells in chronic cervicitis group.The heteromorphic binucleated cells with high nuclear-cytoplasmic ratios and deeply stained nuclei were observed in cervical exfoliated cells of the patients in CIN stage Ⅰ and CIN stage Ⅱ groups.More heteromorphic cells with high nuclear-cytoplasmic ratios and irregular nuclear membranes were showed in cervical exfoliated cells of the patients in CIN stage Ⅲ group.The cervical exfoliated cells of the patients in cervical cancer group showed large and prominent nucleoli,clustering into syncytial changes.Compared with normal group,the atypial of cervical exfoliated cells in CIN stage Ⅰ,CIN stage Ⅱ,CIN stage Ⅲ,and cervical cancer groups was increased.The hybrid capture-chemiluminescence results showed that compared with normal and chronic cervicitis groups,the numbers of HPV infection and TCT heteromorphic cells of the patients in CIN stage Ⅰ,CIN stage Ⅱ,and CIN stage Ⅲ groups were increased(P<0.05);compared with CIN stage Ⅰ,CIN stage Ⅱ,and CIN stage Ⅲ groups,the numbers of HPV infection and TCT heteromorphic cells of the patients in cervical cancer group were increased(P<0.05).The HE staining results showed normal cell morphology and structure in normal group,with infiltration of inflammation cells such as neutrophils,monocytes,macrophages,EOS,and lymphocytes;in chronic cervicitis group,the infiltration of inflammatory cells was increased;in CIN group,the cervical cells showed slightly larger nucleoli and heteromorphic cells,with inflammatory cells mainly distributing around the hetermomorphic cells;in cervical cancer group,the cervical cells showed large and deeply stained nucleoli with significant atypia,and the infiltration of inflammatory cells around the cancer cells was increased.Compared with normal group,the numbers of inflammatory cells and EOS infiltration in cervical tissue of the patients in chronic cervicitis group were increased(P<0.05),and the numbers of inflammatory cells and EOS infiltration of the patients in CIN group were increased(P<0.05);compared with chronic cervicitis group,the number of inflammatory cells and EOS infiltration of the patients in CIN group were decreased(P<0.05);compared with chronic cervicitis group and CIN group,the numbers of inflammatory cells and EOS infiltration of the patients in cervical cancer group were increased(P<0.05).The EOS in cervical cancer tissue was mainly distributed around the cancer nests;compared with CIN stage Ⅰ group,the numbers of EOS infiltration in CIN stage Ⅱ and CIN stage Ⅲ groups were increased(P<0.05);compared with CIN stage Ⅱ group,the number of EOS infiltration in CIN stage Ⅲ group was increased(P<0.05).The higher the malignancy degree of the tumor,the more EOS infiltration was observed,and the number of EOS infiltration was positively correlated with the invasion depth of cervical cancer(r=0.533 0,P<0.01).Conclusion:HPV infection and EOS infiltration play a role in promoting the and occurrence development of cervical precancerous lesions and cervical cancer.
4.Establishment and application of animal models for portal vein thrombosis
Zhuang LIU ; Jihong CHEN ; Xingshun QI ; Xiangbo XU ; Yuzheng ZHUGE
Journal of Clinical Hepatology 2024;40(1):29-32
Portal vein thrombosis (PVT) refers to thromboembolism that occurs in the extrahepatic main portal vein and/or intrahepatic portal vein branches. PVT is the result of the combined effect of multiple factors, but its pathogenesis remains unclear. Animal models are an important method for exploring the pathophysiological mechanism of PVT. Based on the different species of animals, this article reviews the existing animal models of PVT in terms of modeling methods, principles, advantages and disadvantages, and application.
5.Comparison of the clinical efficacy between biopsy forceps polypectomy with submucosal injection and traditional biopsy forceps polypectomy for diminutive colorectal polyps
Qiuli WU ; Yuanli LI ; Chenwei ZHENG ; Xiangbo CHEN ; Qinwei XU
Chinese Journal of Digestive Endoscopy 2024;41(12):979-984
Objective:To explore the clinical efficacy of biopsy forceps polypectomy with submucosal injection for diminutive colorectal polyps.Methods:The patients with diminutive colorectal polyps (long diameter≤5 mm in size) who received polypectomy in Quanzhou First Hospital Affiliated to Fujian Medical University from May 2021 to May 2022 were prospectively recruited and divided into the observation group (biopsy forceps polypectomy with submucosal injection) and the control group (traditional biopsy forceps polypectomy). The location, morphology, long diameter, the complete polypectomy rate under direct vision, the time of resection and endoscopy withdrawal, pathological results, specimen collection rate, the incidence of complications and endoscopic treatment cost were analyzed. The clinical efficacy of the two methods were compared.Results:A total of 292 patients were included in the study, with 146 in the observation group and 146 in the control group. There were 258 polyps in the observation group and 252 polyps in the control group. The complete polypectomy rate was higher in the observation group than that in the control group [100.0% (258/258) VS 90.1% (227/252), χ2=26.915, P=0.001] with clear vision. The incidence of complications in the observation group was lower than that in the control group (1/146 VS 13/146, χ2=10.804, P=0.001) , along with the lower treatment cost (173.7±15.9 yuan VS 184.0±53.8 yuan, Z=-2.777, P=0.005). The resection time in the observation group was longer than that in the control group, but the difference was not statistically significant [10 (9,11) min VS 9 (9,11) min, Z=-0.528, P=0.597]. There was no significant difference in the polyp position, long diameter, morphology, pathological classification, or the specimen collection rate between the two groups ( P>0.05). Conclusion:Biopsy forceps polypectomy with submucosal injection is safe for diminutive colorectal polyps, and it demonstrates a higher complete polypectomy rate, lower complication rates, and reduced treatment costs compared with traditional biopsy forceps polypectomy. It is a new perspective for managing diminutive colorectal polyps.
6.Risk factors for deep vein thrombosis after a spinal cord injury: A retrospective study
Miaoqiao SUN ; Mulan XU ; Xiangbo WU ; Ying LIANG ; Xiao XI ; Yixing LU ; Guiqing CHENG ; Hong WANG ; Ning LI ; Chenguang ZHAO ; Xiaolong SUN ; Hua YUAN
Chinese Journal of Physical Medicine and Rehabilitation 2023;45(4):302-306
Objective:To explore the risk factors for lower extremity deep vein thrombosis (DVT) in patients with a spinal cord injury (SCI).Methods:The medical records of 276 hospitalized SCI patients were analyzed retrospectively. They were divided into a DVT group ( n=63) and a no-DVT group ( n=213). Gender, age, blood type, smoking history, surgical history, the time from SCI to admission, cause of SCI, fracture, SCI segments, American Spinal Cord Injury Association grade and complications were compared between the two groups. Binomial logistic regression was used to isolate the risk factors for lower extremity DVT among such patients. Results:Among 84% of the 63 with a lower extremity DVT, it was a calf muscle venous thrombosis. Anemia, hyponatremia and time from SCI to admission (which ranged from 74 to 195 days) were the most serious DVT risk factors.Conclusions:SCI patients are of high risk for DVT, with anemia and hyponatremia being independent risk factors.
7.Risk factors for urinary tract infection after a spinal cord injury
Yixing LU ; Miaoqiao SUN ; Xiangbo WU ; Mulan XU ; Chunqiu DAI ; Guiqing CHENG ; Wei WANG ; Ying LIANG ; Linna HUI ; Hua YUAN ; Xiaolong SUN
Chinese Journal of Physical Medicine and Rehabilitation 2023;45(5):423-428
Objective:To explore the risk factors for urinary tract infection (UTI) after a spinal cord injury (SCI).Methods:The medical records of 403 SCI patients were analyzed retrospectively. They were divided into UTI group and no-UTI group according to whether they had a UTI at admission. Gender, age, cause of injury, injury level of the spinal cord, voluntary anal contraction, time from injury to admission, American Spinal Injury Association (ASIA) grade, axillary temperature at admission, complications at admission (diabetes, hypertension, fracture of the pelvis, pressure sores or anemia), white blood cell count and urinary bacteria were compared between the two groups. Binary logistic regression was used to highlight the risk factors for a UTI after an SCI.Results:Of the 354 patients included in the final analysis, 62 (17.51%) had a UTI at admission. The regression showed that UTI after an SCI was closely related to an inability to voluntarily contract the anus, anemia, elevated white blood cell count and a high level of bacteria in the urine.Conclusions:Inability to contract the anus, fever, anemia and an elevated white blood cell count are independent indicators of a UTI after an SCI. A temperature ≥37.3°C is a simple indicator of a concentration of bacteria in the urine ≥1266/μL.
8.Risk factors for neuropathic pain after a spinal cord injury: A retrospective study
Mulan XU ; Xiaolong SUN ; Xiangbo WU ; Miaoqiao SUN ; Hong WANG ; Yani ZHANG ; Mi GAO ; Xu HU ; Hui CAO ; Wei SUN ; Chenguang ZHAO ; Hua YUAN
Chinese Journal of Physical Medicine and Rehabilitation 2022;44(3):199-203
Objective:To examine the risk factors for neuropathic pain (NP) after a spinal cord injury (SCI).Methods:A total of 115 patients with a SCI were analyzed retrospectively. They were divided into an NP group of 53 and a non-NP group of 62 according to the occurrence of NP. Gender, age, length of stay, occupation, level of education, cause of injury, spinal fracture, degree of SCI, the injury′s plane and complications at admission (diabetes, hypertension, anemia, venous thrombosis, pressure sores, urinary tract infection or hypoproteinemia) were recorded. T-tests and chi-squared tests were used to compare those factors between the two groups, and multivariate logistic regressions were evaluated to identify the risk factors for NP.Results:Twenty-three of the 53 cases of NP (43%) had developed within 1 month of the SCI. Thirty-seven (75%) experienced pain below the plane of the SCI. The main features reported were squeezing (34%) and numbness (26%). The multivariate logistic regression showed that the occurrence of NP was most strongly related to gender (women being particularly at risk) and venous thrombosis at admission.Conclusions:Women are at particular risk of feeling NP after an SCI, and venous thrombosis is an independent risk factor. NP should be diagnosed and treated quickly to reduce the negative impact on patients′ life quality.
9.Do methylenetetrahydrofolate dehydrogenase, cyclohydrolase, and formyltetrahydrofolate synthetase 1 polymorphisms modify changes in intelligence of school-age children in areas of endemic fluorosis?
Zichen FENG ; Ning AN ; Fangfang YU ; Jun MA ; Na LI ; Yuhui DU ; Meng GUO ; Kaihong XU ; Xiangbo HOU ; Zhiyuan LI ; Guoyu ZHOU ; Yue BA
Chinese Medical Journal 2022;135(15):1846-1854
BACKGROUND:
Excessive exposure to fluoride can reduce intelligence. Methylenetetrahydrofolate dehydrogenase, cyclohydrolase, and formyltetrahydrofolate synthetase 1 ( MTHFD1 ) polymorphisms have important roles in neurodevelopment. However, the association of MTHFD1 polymorphisms with children's intelligence changes in endemic fluorosis areas has been rarely explored.
METHODS:
A cross-sectional study was conducted in four randomly selected primary schools in Tongxu County, Henan Province, from April to May in 2017. A total of 694 children aged 8 to 12 years were included in the study with the recruitment by the cluster sampling method. Urinary fluoride (UF) and urinary creatinine were separately determined using the fluoride ion-selective electrode and creatinine assay kit. Children were classified as the high fluoride group and control group according to the median of urinary creatinine-adjusted urinary fluoride (UF Cr ) level. Four loci of MTHFD1 were genotyped, and the Combined Raven's Test was used to evaluate children's intelligence quotient (IQ). Generalized linear model and multinomial logistic regression model were performed to analyze the associations between children's UF Cr level, MTHFD1 polymorphisms, and intelligence. The general linear model was used to explore the effects of gene-environment and gene-gene interaction on intelligence.
RESULTS:
In the high fluoride group, children's IQ scores decreased by 2.502 when the UF Cr level increased by 1.0 mg/L (β = -2.502, 95% confidence interval [CI]:-4.411, -0.593), and the possibility for having "excellent" intelligence decreased by 46.3% (odds ratio = 0.537, 95% CI: 0.290, 0.994). Children with the GG genotype showed increased IQ scores than those with the AA genotype of rs11627387 locus in the high fluoride group ( P < 0.05). Interactions between fluoride exposure and MTHFD1 polymorphisms on intelligence were observed (Pinteraction < 0.05).
CONCLUSION
Our findings suggest that excessive fluoride exposure may have adverse effects on children's intelligence, and changes in children's intelligence may be associated with the interaction between fluoride and MTHFD1 polymorphisms.
Child
;
Creatinine
;
Cross-Sectional Studies
;
Fluorides/urine*
;
Formate-Tetrahydrofolate Ligase
;
Humans
;
Intelligence/genetics*
;
Methylenetetrahydrofolate Dehydrogenase (NADP)
;
Methylenetetrahydrofolate Reductase (NADPH2)
10.Comparison of the efficacy of different venous intubation in venous-arterial extracorporeal membrane oxygenation assisted lung transplantation
Xiaoliang QIAN ; Yue CHEN ; Li WEI ; Xiangbo JIA ; Lei XU ; Fudong TANG ; Jiaqiang ZHANG ; Peijun REN ; Jianchao LI ; Leiyi YANG ; Zhaoyun CHENG
Chinese Critical Care Medicine 2021;33(9):1080-1083
Objective:To compare the curative effects of different venous cannulas and drainage to improve patient's whole body oxygenation during the auxiliary process of venous-arterial extracorporeal membrane oxygenation (VA-ECMO) in lung transplantation.Methods:From December 2016 to December 2019, 12 patients who were assisted by VA-ECMO in one lung transplantation in People's Hospital of Henan Province were selected as the research objects. According to the number of side holes of venous cannulas, they were divided into two groups: one group with few side holes and other group with multiple side holes. The differences in blood gas indexes among the right radial artery, left radial artery, and right internal jugular vein before and after assistance were compared, and the assistance effect was evaluated.Results:The arterial partial pressure of oxygen (PaO 2) of blood gas indexes of the right and left radial arteries in both groups were significantly higher than that before assistance [mmHg (1 mmHg = 0.133 kPa): right and left radial artery in few side holes group: 79.5±4.2 vs. 48.3±3.8 and 88.1±3.5 vs. 48.3±3.8; right and left radial artery in multiple side holes group: 67.7±5.9 vs. 48.7±3.2 and 84.0±3.8 vs. 48.7±3.2, all P < 0.05]. The arterial partial pressure of carbon dioxide (PaCO 2) of blood gas index was significantly lower than that before assistance (mmHg: 44.2±2.6 vs. 71.7±4.4 for the right radial artery and 44.7±1.4 vs. 71.7±4.4 for the left radial artery in the group with few side holes; 46.2±2.1 vs. 71.2±3.5 for the right radial artery and 44.1±1.9 vs. 71.2±3.5 for the left radial artery in the group with multiple side holes, all P < 0.05). The partial pressure of oxygen in venous blood (PvO 2) of blood gas index of ECMO system in the group with few side holes was significantly lower than that of the multiport side holes group (mmHg: 56.4±3.2 vs. 88.7±1.5, P < 0.01), and the partial pressure of carbon dioxide in venous blood (PvCO 2) was significantly higher than that of multiport side holes group (mmHg: 63.6±3.7 vs. 44.2±1.7, P < 0.01). Conclusions:When VA-ECMO is used in lung transplantation, the superior vena cava blood flow can be fully drained by using intravenous cannula with few side holes. It can effectively improve the oxygenation of the upper body of lung transplant patients, avoid the dilemma of hypoxemia in the upper body and hyperxemia in the lower body, provide more effective assistance to patients undergoing single lung transplantation, and is more meaningful for improving the oxygenation status of the whole body in patients undergoing single lung transplantation.

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