1.Elastase quantitative analysis of elastin effect on mechanical response of anterior cruciate ligament
Wentian ZHANG ; Yuping DENG ; Xiaoyun LIU ; Caijuan LI ; Mian WANG ; Zeyu LIANG ; Liang XIONG ; Gang HUANG ; Guangxin CHEN ; Zitao LI ; Wenhua HUANG
Chinese Journal of Tissue Engineering Research 2024;28(22):3451-3456
BACKGROUND:The anterior cruciate ligament has unique nonlinear mechanical properties under a complex physiological loading environment.Elastin is an important contributor to the mechanical properties of the anterior cruciate ligament,but its mechanical response to the anterior cruciate ligament under axial tension is not clear. OBJECTIVE:To quantitatively analyze the effect of elastin on the tensile mechanical response of the anterior cruciate ligament. METHODS:Elastase solution and control buffer were prepared.The porcine anterior cruciate ligament samples were prepared into small-size samples and randomly soaked in 0,0.1,1.0,2.0,5.0,and 10.0 U/mL elastase solution for 6 hours,and other small samples of the same size were soaked in 2 U/mL elastase solution for 0,1,3,6,9,and 12 hours.To determine suitable soaking conditions for elastin-targeted enzymes and verify the digestive effect,histological staining was used to compare the effects of enzyme treatment on tissue structure and composition.The ligament samples were randomly divided into elastase-treated group and PBS group,and immersed in 2 U/mL elastase solution and PBS buffer for 6 hours,respectively.A mechanical tensile test was performed before and after immersion. RESULTS AND CONCLUSION:(1)The biochemical results showed that being treated in 2 U/mL elastase solution for 6 hours could reduce the elastin content by 31.1%,and had no significant effect on other mechanical-related components in the tissue.(2)The histological results showed that elastase was able to penetrate the tissue,and the loose degree of tissue increased after treatment.(3)In the mechanical results before and after treatment,the mechanical properties of the PBS group decreased significantly,only the low-tension elastic modulus increased significantly and the initial length increased significantly in the elastase-treated group.(4)The intergroup comparison results showed that there was no significant difference between the two groups in pre-treatment,but the low-tension elastic modulus,initial slopes,saturated slopes,and initial length of the elastase-treated group after treatment were significantly higher than those in the PBS group.(5)These results suggest that elastin degradation significantly affects the biomechanical properties of the anterior cruciate ligament and further complements our understanding of the structure-function relationship of the anterior cruciate ligament.
2.The evaluation and influence factors analysis of CT and MRI on thoracolumbar burst fracture combined with PLC injury
Ruibo WANG ; Zitao WANG ; Yanping LI ; Bangjian SUN
China Medical Equipment 2024;21(6):50-55
Objective:To explore the evaluation and influence factors of computed tomography(CT)and magnetic resonance imaging(MRI)on thoracolumbar burst fracture combined with injury of posterior ligament complex(PLC).Methods:A total of 68 patients with thoracolumbar burst fractures who were diagnosed and treated in Handan First Hospital from January 2020 to June 2023 were selected as the research object,and the surgical result was used as gold standard.The 32 cases,who were diagnosed as thoracolumbar burst fractures combined with PLC injury according to the gold standard,were divided into PLC group.The 32 cases without PLC injury were divided into non-PLC group.Before operation,all patients underwent CT and MRI examinations,and the positively and negatively predictive values of CT and MRI in diagnosing thoracolumbar burst fracture combined with PLC injury were calculated by four-grid method.The area under curve(AUC)value,sensitivity and specificity of CT and MRI in diagnosing thoracolumbar burst fracture combined with PLC injury were analyzed by receiver operating characteristic(ROC)curve.Logistic regression model was used to analyze the risk factors of patients with thoracolumbar burst fracture who occurred PLC injury.The differences of the scores of ligamentous complex stability(LCS)score,intraspinal space occupancy rate and thoracolumbar injury classification and severity(TLICS)score,and the scoliosis angle(Cobb),superior iliac crest angle(SIEA),local kyphosis(LK)angle and intervertebral disc space depth(IISD)between two groups were compared.Results:For 68 with thoracolumbar burst fractures,the 34 cases were confirmed as PLC injury and 34 cases were confirmed as non-PLC injury by using CT examination.The positively and negatively predictive values of CT examination were respectively 70.59%(24/34)and 76.47%(26/34)for PLC injury,and the consistency between CT and gold standard was general(Kappa=0.471,P<0.001).The 33 cases were confirmed as PLC injury and 35 cases were confirmed as non-PLC injury by using MRI examination.The positively and negatively predictive values of MRI examination were respectively 90.91%(30/33)and 94.29%(33/35)for PLC injury,and the consistency between MRI examination and gold standard was general(Kappa=0.853,P<0.001).The diagnostic accuracy of MRI was 92.65%(63/68),which was significantly higher than that(73.53%,50/68)of CT(x2=8.843,P<0.05).ROC curve analysis showed that the AUC values of CT and MRI were respectively 0.730 and 0.919 in diagnosing thoracolumbar burst fracture combined with PLC injury.The sensitivities of them were respectively 70.60%and 75.40,and the specificities of them were respectively 88.20%and 95.70%.There were no significant differences between PLC group and non-PLC group in gender,age,body mass index(BMI),cause of injury,LCS score and intraspinal space occupancy rate(P>0.05).There were significant differences in TLICS score,Cobb angle,SIEA,LK and IISD between the two groups(x2=19.443,4.181,4.973,5.198,5.056,P<0.05),respectively.Logistic regression analysis showed that TLICS score>5 points,Cobb angle,SIEA,LK and IISD were risk factors that affected the occurrence of PLC injury in patients with thoracolumbar burst fracture(OR=13.973,1.155,1.365,1.385,5.262,P<0.001),respectively.Conclusion:The efficiency of MRI is higher than that of CT in diagnosing PLC injury in patients with thoracolumbar burst fracture,and TLICS score,Cobb angle,SIEA,LK and IISD have influences on the occurrence of PLC injury in patients with thoracolumbar burst fracture.
3.Clinical characteristics and risk factors for preoperative anemia in patients with adult femoral shaft fracture
Weihao MENG ; Xi CHEN ; Xiao MENG ; Xiwen QIAN ; Fengfeng LI ; Zitao ZHANG
Chinese Journal of Orthopaedics 2023;43(20):1379-1386
Objective:To investigate the clinical characteristics and predisposing factors associated with preoperative anemia in adult femoral shaft fractures.Methods:A retrospective analysis of clinical data from 157 patients presenting with femoral shaft fractures admitted to the department of orthopedics at Nanjing Drum Tower Hospital between June 2018 and June 2022 was conducted. The study cohort comprised 106 males and 51 females, with an average age of 45.06 ± 14.32 years (range: 18-65 years). Based on hemoglobin levels measured within 2 days of admission, patients were stratified into two groups: anemia group (Hb<120 g/L in adult males and Hb<110 g/L in adult females) and non-anemia group. General demographic information, AO fracture types, and clinical characteristics, as well as the results of laboratory examinations for both groups were collected. Subsequently, univariate and multivariate logistic regression analyses were conducted.Results:Out of the 157 patients with femoral shaft fractures, 118 (75.2%) exhibited preoperative anemia (the anemia group). Among them, 75 cases were male, and 43 cases were female, with an average age of 45.84±14.23 years (range: 18-65 years). In terms of fracture AO type, 41 cases were classified as 32A, 19 as 32B, and 58 as 32C. Regarding fracture location, 14 were situated in the upper 1/3 of the femoral shaft, 67 in the middle 1/3, and 37 in the lower 1/3. The causes of injury included 63 cases of motor vehicle accidents, 5 cases of blunt trauma, 40 cases of falls, and 10 cases of other falls, with 65 cases involving multiple injuries. Conversely, 39 patients (24.8%) did not exhibit preoperative anemia (the non-anemia group). Of these, 31 were male, and 8 were female, with an average age of 42.72 ± 14.51 years (range: 19-65 years). In terms of fracture AO type, 24 cases were classified as 32A, 5 as 32B, and 10 as 32C. Regarding fracture location, 3 were situated in the upper 1/3 of the femoral shaft, 19 in the middle 1/3, and 17 in the lower 1/3. The causes of injury included 13 cases of motor vehicle accidents, 5 cases of blunt trauma, 20 cases of falls, and 1 other fall, with 8 cases involving multiple injuries. Univariate analysis revealed statistically significant differences between preoperative anemia and AO fracture type, mechanism of injury, multiple injuries, time from injury to hospital admission, albumin levels, and age ( P< 0.05). Multifactorial logistic regression analysis identified AO type 32C ( OR=3.12, P=0.020), blunt trauma injuries ( OR=0.13, P=0.021), reduced albumin levels ( OR=9.90, P=0.037), and multiple injuries ( OR=3.65, P=0.016) as risk factors for preoperative anemia. Multifactorial logistic regression further revealed that multiple injuries ( OR=5.20, P=0.004) and reduced albumin levels ( OR=5.47, P=0.001) were risk factors for the severity of anemia. Conclusion:AO type 32C fractures, blunt trauma injuries, reduced albumin levels, and multiple injuries were identified as potential contributors to the development of preoperative anemia, with multiple injuries and reduced albumin levels exacerbating the severity of anemia. Clinicians should be vigilant for the occurrence of preoperative anemia in adult femoral shaft fracture patients, particularly those with blunt trauma injuries, multiple injuries, hypoalbuminemia, and AO type 32C fractures.
4.Effect of carpesium abrotanoides petroleum ether fraction on phagocytic function of macrophages and antibody forming cell in mice
Li LIN ; Yudan ZHOU ; Jiaqi XU ; Zitao WANG ; Shuo XIAO ; Guoxiang ZHOU ; Jiang ZHENG ; Qiang WANG
International Journal of Traditional Chinese Medicine 2019;41(5):487-490
Objective To study the effects of carpesium abrotanoides petroleum ether fraction on phagocytic function of macrophages and antibody forming cell in mice.Methods The extract carpesium abrotanoides petroleum ether fraction,forty five mice were divided into the control group and low and high dose groups of carpesium abrotanoides petroleum ether fraction by random number table method,15 mices in each group.The low-dose and high-dose groups of carpesium abrotanoides petroleum ether fraction were given 50 mg/kg and 100 mg/kg respectively by gavage for 10 days.The spleen weight,growth index,the phagocytosis of macrophages and the number of antibody-forming cells in peritoneal macrophages of mice were detected by chicken erythrocyte phagocytosis test and hemolytic plaque test.Results Compared with the control group,the spleen weight (192.4 ± 11.49 mg,204.6 4 10.59 mg vs.117.6 ± 10.89 mg),the growth index (6.04 ± 0.54,6.06 ± 0.40 vs.3.89 ± 0.14),antibody forming (1 216.4 ± 94.1,1 548.8 ± 86.4 vs.361.0 ± 11.7),phagocytosis percentage of macrophages (58.60% ± 2.60%,72.0% ± 3.08% vs.35.49% ± 1.64%),and Phagocytosis index (2.01 ± 0.10,2.69 ± 0.15 vs.0.37 ± 0.06) of the groups of low doses and high doses of Carpesium abrotanoides Petroleum ether fraction significantly increased (P<0.05).Conclusions The low doses and high doses of Carpesium abrotanoides Petroleum ether fraction can enhance the mice's specific and nonspecific immune function,and protect the immune system of mice.
5.Anti-tumor effect and its mechanism of co-administration of fusion proteins hVEGF121/βhCG and mGM-CSF/βhCG
Liangliang JING ; Zitao MIAO ; Manman LI ; Jia YE ; Liang JIN ; Rongyue CAO ; Jun LONG
Journal of China Pharmaceutical University 2017;48(1):102-109
This study aimed at investigating the inhibitory effects and the anti-tumor mechanisms of co-adminis-tration of fusion proteins mGM-CSF/βhCG ( GC ) and hVEGF121/βhCG ( VC ) on RM-1 prostatic cancer and B16 F10 melanoma in the C57 BL/6 J mouse model. Two recombinant stains containing pET-28 a-mGM-CSF-X10-βhCGCTP37 and pET-28 a-VEGF-M2-X10-βhCG-CTP37 were induced by lactose to express fusion proteins. The fusion proteins were separated and purified to prepare the anti-tumor protein vaccines ( VC protein vaccine and GC protein vaccine) , which were then mixed to prepare a combined protein vaccine named VGC protein vac-cine. The prostatic cancer and melanoma tumor-bearing mice C57 BL/6 J were immunized with described vac-cines, then the growth of each tumor was measured;splenocyte proliferation of immunized mice was detected;and the cytotoxic effects of the vaccine on tumor cells were tested. After that, the in vivo concentrations of IFN-γ and anti-hVEGF antibodies were investigated by ELISA. The difference between each experimental group and normal saline group ( NS) was statistically significant in both tumor-bearing mouse models ( P <0. 05) respectively. Besides, VGC group exhibited significantly better anti-tumor effect compared with the GC and VC groups with the anti-tumor rate ( 41. 7 ± 0. 83)% and ( 46. 4 ± 1. 27)% for prostatic cancer and melanoma tumor, respectively. The co-administration of the two proteins, VC and GC, could inhibit the growth of RM-1 prostatic tumor and B16F10 melanoma effectively via anti-tumor immunity and anti-tumor angiogenesis.
6.Effects of Mycobacterium tuberculosis HSP65 on Treg/Th17 immune balance in ApoE-knockout mice
Rongyue CAO ; Xinli ZHANG ; Dongping YUAN ; Manman LI ; Minxia YU ; Yunfei MA ; Zitao MIAO ; Jun LONG
Journal of China Pharmaceutical University 2016;47(3):353-358
To investigate the effects of Mycobacterium tuberculosis heat shock protein 65(HSP65)on Treg/Th17 immune balance in ApoE-knockout(ApoE-/-)mice, ApoE-/- mice with a high-cholesterol diet were immunized with M. tuberculosis HSP65. Sera were obtained for measurement of anti-HSP65 antibodies by ELISA; the effect of administration of different antigens was investigated, respectively, using flow cytometry analysis on the number of CD4+CD25+Foxp3+Tregs and CD4+IL-17+ Th17; the production of cytokines(IL-10, TGF-β1, IL-17 and IL-21)by these cells were determined by ELISA; total plasma cholesterol(TC), triglyceride(TG), high-density lipoprotein cholesterol(HDL-C)and low-density lipoprotein cholesterol(LDL-C)levels were detected by biochemical autoanalyzer. Atherosclerotic lesions were measured by lipid deposition stained with oil red O. The results demonstrated that the levels of anti-HSP65 IgG antibodies were increased significantly in Mycobacterium tuberculosis HSP65-treated ApoE-/- mice, revealed obvious decrease in Treg number, Treg related cytokines(IL-10, TGF-β1)levels and significant increase in Th17 number, Th17 related cytokines(IL-17 and IL-21)levels, the levels of TC, TG, HDL-C and LDL-C did not change between groups, while the atherosclerotic lesions significantly increased. Results indicate that M. tuberculosis HSP65 could interrupt the Th17/Treg immune balance in ApoE-/- mice, suggesting a potential role in the formation and progression of atherosclerosis.
7.Costimulate moleculars amplified the DRibble mediated T cells response
Yun XING ; Zhenxian ZHOU ; Zitao MIAO ; Manman LI ; Rongyue CAO ; Jun LONG
Journal of China Pharmaceutical University 2016;47(6):749-754
Autophagosomes derived from tumor cells have been proved to induce potent T cell response both in mouse and human. In human in vitro study, dendritic cells(DC)loaded with cytomegalovirus(CMV)pp65 antigen-containing DRibble were capable to efficiently re-stimulate pp65-specific T-cell recall responses from freshly isolated or frozen humanperipheral blood mononuclear cell(PBMC). This study developed more robust assays using in vitro expanded antigen-specific T cells that contained a much higher percentage of antigen-specific T cells. DC cross-presentation efficiency of OX40 and CD80 modified pp65-DRibble was detected by intracellular IFN-γ staining. Compared with Ctrl/pp65 DRibble, the percentage of IFN-γ+ in total CD8+ T cells andCD4+ T cells was improvedwith OX40/pp65 DRibbleand CD80/pp65 DRibble stimulation. In addition, vaccine induced IL-12indendritic cells, whichpolarizes Th cells toward the IFN-γ high Th1 phenotype, evaluated by ELISA inco-culture supernatantwas dramatically higher in OX40/pp65 DRibble and CD80/pp65 DRibblegroups than in Ctrl/pp65 DRibble group. These results have implications for the immuneactivity of OX40 and CD80 modified DRibble and choice for prospective clinical use ofDRibble-based cancer immunotherapy.
8.The predictive value of semen parameters in intrauterine insemination cycles of male factors infertile couples
Zonghui XIAO ; Jiaying LIANG ; Zitao LI ; Peng DU ; Xiaoxuan ZHENG ; Lihu WANG ; Fenghua LIU
International Journal of Laboratory Medicine 2015;(23):3374-3377
Objective To confirm the correlation between pre or post processing semen parameters and clinical pregnancy rate in infertile couples with male factors undergoing intrauterine insemination (IUI) .Methods The pregnancy rate based on ovulation in‐ducing agent in infertile couples with male factors infertility per the 2010 world health organization criteria treated with IUI was e‐valuated .One hundred and twenty eight cycles were divided into pregnant group (16 cycles) and non‐pregnant group(112 cycles) . The pre or post processing semen parameters were retrospectively evaluated .Results The overall pregnancy rate was 12 .5% .All parameters in this study has no statistically significant difference between the two groups (P>0 .05) .Logistic multivariate analysis showed that there was no significant correlation between semen parameters and clinical pregnancy rate (P> 0 .05) .Conclusion There are no significant correlations between semen parameters and clinical pregnancy rate when the normal forms of patients with male infertility was 4% or higher .No semen parameters could predict results of clinical pregnancy .
9.Non-contrast-enhanced Magnetic Resonance Angiography of the Hepatic Portal Vein at 3.0 Tesla
Feifei YAO ; Jingliang CHENG ; Zitao YANG ; Ying LI ; Guoguo LU
Chinese Journal of Medical Imaging 2013;(11):853-856
Purpose To explore the methods of selectively visualizing hepatic portal vein by using three-dimensional fast imaging employing steady state acquisition combined with in-flow inversion recovery labeling pulse at 3.0 Tesla. Materials and Methods Ten healthy volunteers were examined under different TI (1200, 1400, 1600, 1800 ms), and the vessel-to-liver contrast ratio of the main portal vein, right portal vein, and left portal vein were measured. Results Non-contrast-enhanced MRA images of portal vein were obtained successfully in all ten volunteers. The signal intensity of peripheral portal branches gradually increased when TI increased from 1200 ms to 1600 ms, and the highest vessel-to-liver contrast ratio occurred when TI was 1400 ms. Conclusion Non-contrast enhanced magnetic resonance angiography of the hepatic portal vein can be successfully achieved at 3.0T high field MRI. A fixed TI of 1400 ms is preferable.
10.Opportunistic screening for colorectal cancer
Zitao WU ; Jianqiu SHENG ; Li ZHANG ; Dongxu WANG ; Bingxin WANG ; Libo LIU ; Jian ZHU ; Ying HAN
Chinese Journal of Health Management 2012;06(5):318-321
Objective To retrospectively analyze medical data of patients with colorectal cancer (CRC) so as to provide evidence for clinical use of opportunistic screening.Methods A total of 2450 CRC patients (male 1377,female 1073) who were treated at five hospitals in North China during October 2001 and September 2011 and had complete medical records and pathological results were recruited.The correlations of incidenceofCRCwithage,gender,tumorlocationandhistologicaltypeswere analyzed.Results Of all the CRC patients,those less than 50 years old accounted for 18.14% ; and the incidence of CRC was substantially increased in those over 50 years old.Seventy-three percent of tumor occurred at the rectum and sigmoid colon,6% at descending colon,7% at transverse colon and 14% at ascending colon.Moderately,well or poorly differentiated adenocarcinoma accounted for 50.33%,40.35%and 9.32%,respectively.Histological differentiation was not correlated with age and gender ( P > 0.05 ).Conclusions Age and gender should not be considered a determination of opportunistic screening for CRC.Colonoscopy is recommended as an alternative CRC screening procedure.

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