BACKGROUND:Bone marrow mesenchymal stem cel s and al ogenic bones are commonly used as seed cel s and scaffolds, respectively, for constructing tissue-engineered cartilage through in vitro co-culture. The loose body of the knee joint can survive in the articular cavity for a long time, and maintain certain characteristics of cartilage tissues. Therefore, the articular cavity can provide a good environment for the growth and development of chondrocytes. OBJECTIVE:To investigate the effect of bone marrow mesenchymal stem cel s co-cultured with al ogenic bone in the articular cavity. METHODS:Bone marrow mesenchymal stem cel s were isolated from five newborn New Zealand white rabbits. One adult New Zealand rabbit was enrol ed to prepare al ogenic bone for co-culture with bone marrow mesenchymal stem cel s. Afterwards, bone marrow mesenchymal stem cel s and al ogenic bone composites were cultured in the articular cavity (intracavitary culture group) or in vitro as in vitro culture group, respectively;the normal cartilage tissues grew in the articular cavity as control group. Cel s were observed by hematoxylin-eosin staining and type II col agen immunohistochemistry staining at 4, 8 and12 weeks of culture. RESULTS AND CONCLUSION:At 12 weeks culture, hematoxylin-eosin staining showed:in the control group, chondrocytes arranged tightly and directional y with red stained cytoplasm and cartilage matrix as wel as blue nuclei;in the intracavitary culture group, the scaffold was mostly absorbed and chondrocytes grew into the scaffold in a certain direction with smaller shape, while cytoplasm and cartilage ma trix were red stained, blue nuclei appeared;in the in vitro culture group, abundant chondrocytes proliferated in a disordered arrangement. Immunohistochemistry staining showed:the absorbance ( A) values in the intracavitary culture group showed a continuous increase, but no obvious change was in the other two groups. Moreover, at 4, 8 and 12 weeks of culture, A values in the control group and intracavitary culture group were significantly higher than that in the in vitro culture group (P<0.05);at 4 and 8 weeks, A value in the intracavitary culture group was significantly lower than that in the control group (P<0.05), but at 12 weeks, there was no significant difference in A value between the two groups (P>0.05). These results suggest that the tissue-engineered cartilage can be constructed by bone marrow mesenchymal stem cells co-cultured with allogenic bone under in vitro and in vivo environment, especially in the articular cavity.

Objective To investigate the role of bacterial flagellin and CD98 in ulcerative colitis (UC).Methods A total of 60 first episode patients with active UC were recruited,including 30 mild and 30 moderate to severe UC cases.The serum of 30 healthy volunteers and normal intestinal tissues surgically removed from 15 colon cancer patients (more than 5 cm away from surgical margins) were collected as control.The content of bacterial flagellin antibodies in peripheral blood were measured with enzyme-linked immunosorbent assay (ELISA).The expression of CD98 in peripheral blood T lymphocyte was measured by flow cytometry (FACS).The expression of bacterial flagellin protein in intestinal mucosa and CD98 in intestinal epithelial basement membrane was tested by immunohistochemistry (IHC).The comparison between two groups was performed with the SNK-q method,the R×C table x2 test was used to analyze the counted data,and the Spearman correlation was used to analyze the rank materials.Results The peripheral blood concentration of bacteria flagella protein antibody of control group,mild UC group and moderate to severe group showed an upward trend,which was (7.603±2.118) pg/ml,(13.702±3.131) pg/ml and (20.813±3.004) pg/ml respectively,and the differences among groups were statistically significant (F=13.57,P＜0.01).The expression percentage of bacteria flagella protein in intestinal mucosa of the three groups also showed an upward trend,which was 3/15,56.67％ and 73.33％ respectively,and the differences among groups were statistically significant (x2 =11.553,P=0.003).The positive rate of CD98 expression in peripheral blood T lymphocytes of the three groups showed an upward trend,which was (28.42±4.31)％,(32.45±6.71)％ and (43.40±5.09) ％ respectively,and the differences among groups were statistically significant (x2 =10.110,P=0.007).The positive rate of CD98 expression in intestinal epithelial cells of the three groups also showed an upward trend,which was 1/15,36.67 ％ and 66.67％ respectively,and the differences among groups were statistically significant (x2 =5.400,P＜0.05).There was positive correlation between the peripheral blood concentration of bacteria flagella protein antibody and the expression of CD98 in peripheral blood T lymphocytes (r=0.548,P＜0.05).Conclusion Bacterial flagellin and CD98 may be important factors causing inflammatory reaction activity in UC.

Objective To evaluate the diagnosis and application values of endoscopic ultrasonography (EUS) with micro-probe in duodenal lesions.Methods Clinical data of 37 patients with duodenal lesions and underwent EUS with micro-probe were analyzed retrospectively.All lesions were treated with endoscopic mucosal resection or surgical resection to get the pathological diagnosis.The diagnostic accuracy of EUS with microprobe and endoscopic biopsy was analyzed respectively.Results The overall diagnosis accuracy of EUS with micro-probe on duodenal lesions was 78.38％ (29/37),with a higher diagnostic rate in duodenal lipoma 4/4and duodenal adenomas 10/12 than in early duodenal cancer 2/4 or inflammatory hyperplasia 3/8.The overall diagnostic accuracy of biopsy on duodenal lesions was 40.54％ (15/37),with a higher diagnosis rate on duodenal carcinoid 1/1 and adenoma 7/12 than on duodenal stromal tumor 1/10 and lipoma 1/4.Conclusion Pathological evaluation of endoscopic biopsy sample is not a golden standard for the diagnosis of duodenal lesions,while EUS with micro-probe has better diagnostic and application value.

Inflammatory bowel disease(IBD)is a disease characterized by chronic,recurrent intestinal inflammation,includ-ing Crohn's disease(CD)and ulcerative colitis(UC).Etiology and pathogenesis of IBD are unknown,and the treatment is difficult.In recent years,immunocellular therapy represented by regulatory T(Treg)cell therapy and dendritic cell(DC)therapy has developed rapidly in the field of immune-related disease treatment,and some progress has also been made in the treatment of IBD.This review will introduce application and progression of Treg cell therapy and DC therapy in IBD treatment.

Background: Matrix metalloproteinase-7 (MMP-7) is a proteolytic enzyme involved in wound healing, tissue remodeling by regulating a variety of extracellular matrix and non-matrix substrates, and is an important mediator of tissue damage during inflammation in inflammatory bowel disease (IBD). Aims: To investigate the clinical significance of MMP-7 in IBD. Methods: A total of 38 cases of colon biopsy samples from patients with IBD from January 2019 to January 2021 at the Second Affiliated Hospital of Zhengzhou University were collected, including 20 ulcerative colitis (UC) and 18 Crohn's disease (CD). The paracancerous tissues of 5 patients with colon cancer were served as the control group. The expression of MMP-7 in colon tissue was detected by immunohistochemical staining, and its correlation with C-reactive protein (CRP) and white blood cell (WBC) was analyzed. Results: Compared with the control group, the expression of MMP-7 was significantly up-regulated in both active UC group and severe CD group (P<0.05). The expression of MMP-7 was gradually up-regulated with the increase of severity. The expression of MMP-7 in severe UC and CD group was significantly increased than that in remission UC and CD group (P<0.05). No significant difference in the expression of MMP-7 was found between patients with different gender, age, and whether treated with anti-TNF-α agents (P>0.05). The expression of MMP-7 in colon tissue of patients with active IBD was positively correlated with CRP and WBC (P<0.001). Conclusions: The expression of MMP-7 is correlated with the activity and severity of IBD, but not correlated with gender, age, and whether treated with anti-TNF-α agents. To some extent, MMP-7 can indicate the severity of intestinal inflammation, and may be an important indicator of inflammatory activity in IBD.

Background::Histological healing is closely associated with improved long-term clinical outcomes and lowered relapses in patients with ulcerative colitis (UC). Here, we developed a novel diagnostic criterion for assessing histological healing in UC patients.Methods::We conducted a retrospective cohort study in UC patients, whose treatment was iteratively optimized to achieve mucosal healing at Shanghai Tenth People’s Hospital of Tongji University from January 2017 to May 2022. We identified an inflammatory cell enumeration index (ICEI) for assessing histological healing based on the proportions of eosinophils, CD177 + neutrophils, and CD40L + T cells in the colonic lamina propria under high power field (HPF), and the outcomes (risks of symptomatic relapses) of achieving histological remission vs. persistent histological inflammation using Kaplan-Meier curves. Intrareader reliability and inter-reader reliability were evaluated by each reader. The relationships to the changes in the Nancy index and the Geboes score were also assessed for responsiveness. The ICEI was further validated in a new cohort of UC patients from other nine university hospitals. Results::We developed an ICEI for clinical diagnosis of histological healing, i.e., Y = 1.701X 1 + 0.758X 2 + 1.347X 3 - 7.745 (X 1, X 2, and X 3 represent the proportions of CD177 + neutrophils, eosinophils, and CD40L + T cells, respectively, in the colonic lamina propria under HPF). The receiver operating characteristics curve (ROC) analysis revealed that Y <-0.391 was the cutoff value for the diagnosis of histological healing and that an area under the curve (AUC) was 0.942 (95% confidence interval [CI]: 0.905-0.979) with a sensitivity of 92.5% and a specificity of 83.6% ( P <0.001). The intraclass correlation coefficient (ICC) for the intrareader reliability was 0.855 (95% CI: 0.781-0.909), and ICEI had good inter-reader reliability of 0.832 (95% CI: 0.748-0.894). During an 18-month follow-up, patients with histological healing had a substantially better outcome compared with those with unachieved histological healing ( P <0.001) using ICEI. During a 12-month follow-up from other nine hospitals, patients with histological healing also had a lower risk of relapse than patients with unachieved histological healing. Conclusions::ICEI can be used to predict histological healing and identify patients with a risk of relapse 12 months and 18 months after clinical therapy. Therefore, ICEI provides a promising, simplified approach to monitor histological healing and to predict the prognosis of UC.Registration::Chinese Clinical Trial Registry, No. ChiCTR2300077792.