Objective To construct an injectable hydrogel(IH)entrapment system of bone marrow mesenchymal stem cells(BM-MSCs)and to explore its effect of promoting articular cartilage repair and underlying mechanism.Methods The primary BM-MSCs of mice were cultured by whole bone marrow adherent method.The differentiation potential was identified by alizarin red(ARS),oil red O(ORO)and Alcian blue(AB)staining,and the expres-sion of CD44,CD90,CD105,CD34 and CD45 on the surface was detected by flow cytometry.BM-MSCs loaded with IH(BM-MSC-IH)was prepared,and the rheological properties of BM-MSC-IH were tested by rheological test.The model of full-thickness injury of knee cartilage in mice was established,and the mice were randomly divided into normal saline group,BM-MSC group and BM-MSC-IH group with 8 in each.The mice were injected with BM-MSC-IH 0.2 mL,cell suspension 0.2 mL and 0.9％NaCl solidion,independently.After 6 and 12 weeks of in-jection,the repair of knee cartilage in mice was observed by safranin-fast green staining,and the expression of typeⅡ collagen in knee joint was examined by immunohistochemical staining microscopy.Results The primary cultured cells showed positive expression of CD44,CD90 and CD105,but negative expression of CD34 and CD45,and had the potential of osteogenic,adipogenic and chondrogenic differentiation.The constructed BM-MSC-IH sys-tem was solid colloid,and the rheological properties were consistent with the basic characteristics of hydrogel.BM-MSC-IH effectively promoted the regeneration of mouse knee cartilage and increase the local expression of typeⅡ collagen.Conclusions Bone marrow-mesenchymal stem cells embedded with injectable hydrogel effectively pro-mote the repair of injured cartilage and the underlying mechanism is potentially promoting the differentiation of chondrocytes and the expression of type Ⅱ collagen.Its effect is better than application of BM-MSCs alone.

Glioma is the most common primary malignant brain tumor and its microenvironment exhibits immunosuppressive properties. Macrophages and T cells are the main immune cells of glioma, which engage in highly dynamic interactions. Cytokines such as interleukin-2 (IL-12), interleukin-10 (IL-10), interferon-γ (IFN-γ),and transforming growth factor-β (TGF-β) determine the direction and intensity of the anti-tumor immune response through finely regulating macrophage polarization and T cell subset differentiation. Co-stimulatory molecules on the surface of T cells are mostly members of the immunoglobulin superfamily (IgSF); in addition,co-stimulatory molecules including CD80/CD86, T cell inducible co-stimulatory molecule and its ligand (ICOS /ICOS-L),CD40L/CD40, OX40L/OX40 and glucocorticoid-induced tumor necrosis factor receptor related protein and its ligand (GITR /GITR-L) are involved in initiating,enhancing or inhibiting T cell activation,and collaboratively shape the overall tumor immune microenvironment. The in-depth understanding of these factors and molecular pathways can help optimize immunotherapeutic strategies for glioma and provide new therapeutic targets.

Objective To explore the clinical application value of contrast-enhanced ultrasound in the differential diagnosis of testicular masses.Methods A total of 63 patients at the First Affiliated Hospital of Wenzhou Medical University with testicular masses from January 2014 to August 2024 were sellected.All patients underwent both conventional ultrasound and contrast-enhanced ultrasound examinations.Among them,pathological results were obtained through surgical resection in 55 cases,and 8 non-tumorous patients were diagnosed through clinical follow-up.According to the results,testicular masses were classified into neoplastic and non-neoplastic lesions.Neoplastic lesions were further divided into benign and malignant masses.The routine ultrasound parameters,serum tumor markers and ultrasound contrast parameters were analyzed.Results In the differential diagnosis between neoplastic and non-neoplastic lesions of the testis,there were significant statistical differences in the maximum diameter of the mass,blood flow grade,enhancement time,and enhancement intensity(P<0.05).In the differential diagnosis between benign and malignant testicular masses,significant statistical differences were found in mass location,maximum diameter of the mass,echo characteristics,blood flow grade indicators,enhancement time,and enhancement intensity(P<0.05).Conclusion Contrast-enhanced ultrasound may have certain reference value in the diagnosis of testicular masses.Hypo-enhancement or non-enhancement,along with slow enhancement,may be manifestations of benign lesions.

Objective:To compare the effectiveness between the unilateral biportal endoscopy (UBE) and the bilateral biportal endoscopy (BBE) decompression in the treatment of two-level central lumbar spinal stenosis (LSS).Methods:From January 2022 to April 2024, the clinical data of 31 patients with two-level central LSS treated with UBE and BBE unilateral approach with bilateral decompression were retrospectively analyzed. There were 17 males and 14 females; the age ranged from 60 to 82 years, with a mean of (71.2±5.9) years. The operative segments were L 2-3 and L 3-4 in 2 cases, L 3-4 and L 4-5 in 29 cases. Among them, 15 cases were treated with UBE and the other 16 cases were treated with BBE. The age, gender, course of disease, operation time, intraoperative fluoroscopy frequency, ambulation time, hospitalization days, incision healing grade and surgical complications were compared between the two groups. Visual analogue scale (VAS) and Oswestry disability index (ODI) were used to assess the low back and leg pain degree and functional improvement situation before operation, 3 months after operation and at last follow-up. Imaging examinations were performed before and after operation to evaluate the height of intervertebral space, the rate of articular process preservation and the area of dural sac in the two groups. Measurement data with normal distribution were represented as mean±standard deviation( ± s), and the comparison between groups was conducted using the t-test; measurement data with skewed distribution were represented as (interquartile range) [ M( Q1, Q3)], inter-group comparisons were conducted using the two-sample rank sum test, and intra-group comparisons before and after surgery were conducted using the rank sum test for two related samples or the rank sum test for multiple related sample data. The count data were represented as cuses and percentages, and the comparison between groups was conducted using the Chi-square test or Fisher exact probability method. Results:Thirty-one patients were successfully operated and followed up for 6-18 months, with an average follow-up time of (11.4±3.2) months. There was no significant difference in age, gender, course of disease, ambulation time and hospitalization days between the two groups ( P>0.05). There were significant differences between UBE and BBE in fluoroscopy frequency [(4.2±0.7) vs (2.3±0.4)] and operation time [(118.2±12.8) min vs (72.3±5.6) min] ( P<0.001). Three months after operation and at last follow-up, the VAS scores and ODI were significantly lower than that befor the operation, and the dural sac area was significantly larger than that before the operation in the two groups ( P<0.001), but there was no significant difference in VAS, ODI and dural sac area before or after operation between the two groups ( P>0.05). There was no statistical difference in the intervertebral height between the two groups compared to their respective preoperative measurements( P>0.05). The rate of articular process preservation on the operated side was about 80% in both groups. There were no complications such as dural nerve injury and hemorrhage in both groups. One patient in the UBE group had incision infection, which was improved after symptomatic treatment. Conclusions:Both UBE and BBE can achieve satisfactory effectiveness in the treatment of two-level central LSS, and the clinical effectiveness is similar. BBE can improve the operation efficiency, shorten the surgical duration and reduce the fluoroscopy frequency, so it has more advantages in the treatment of two-level central LSS.

Objective To explore the clinical application value of contrast-enhanced ultrasound in the differential diagnosis of testicular masses.Methods A total of 63 patients at the First Affiliated Hospital of Wenzhou Medical University with testicular masses from January 2014 to August 2024 were sellected.All patients underwent both conventional ultrasound and contrast-enhanced ultrasound examinations.Among them,pathological results were obtained through surgical resection in 55 cases,and 8 non-tumorous patients were diagnosed through clinical follow-up.According to the results,testicular masses were classified into neoplastic and non-neoplastic lesions.Neoplastic lesions were further divided into benign and malignant masses.The routine ultrasound parameters,serum tumor markers and ultrasound contrast parameters were analyzed.Results In the differential diagnosis between neoplastic and non-neoplastic lesions of the testis,there were significant statistical differences in the maximum diameter of the mass,blood flow grade,enhancement time,and enhancement intensity(P<0.05).In the differential diagnosis between benign and malignant testicular masses,significant statistical differences were found in mass location,maximum diameter of the mass,echo characteristics,blood flow grade indicators,enhancement time,and enhancement intensity(P<0.05).Conclusion Contrast-enhanced ultrasound may have certain reference value in the diagnosis of testicular masses.Hypo-enhancement or non-enhancement,along with slow enhancement,may be manifestations of benign lesions.

Glioma is the most common primary malignant brain tumor and its microenvironment exhibits immunosuppressive properties. Macrophages and T cells are the main immune cells of glioma, which engage in highly dynamic interactions. Cytokines such as interleukin-2 (IL-12), interleukin-10 (IL-10), interferon-γ (IFN-γ),and transforming growth factor-β (TGF-β) determine the direction and intensity of the anti-tumor immune response through finely regulating macrophage polarization and T cell subset differentiation. Co-stimulatory molecules on the surface of T cells are mostly members of the immunoglobulin superfamily (IgSF); in addition,co-stimulatory molecules including CD80/CD86, T cell inducible co-stimulatory molecule and its ligand (ICOS /ICOS-L),CD40L/CD40, OX40L/OX40 and glucocorticoid-induced tumor necrosis factor receptor related protein and its ligand (GITR /GITR-L) are involved in initiating,enhancing or inhibiting T cell activation,and collaboratively shape the overall tumor immune microenvironment. The in-depth understanding of these factors and molecular pathways can help optimize immunotherapeutic strategies for glioma and provide new therapeutic targets.

Objective:To explore the relationship between the transforming growth factor-β (TGF-β) signaling pathway and steroid-induced osteonecrosis of the femoral head in young rabbits.Methods:Sixty 8-week-old rabbits weighing 1.5-2.0 kg were randomly divided into steroid injection group (48 cases) and control group (12 cases). Rabbits in the former group were injected with Prednisolone Acetate 7.5 mg/kg into bilateral gluteal muscles twice a week for 8 weeks, and those with successful modeling were included in the disease group; otherwise, they were included in the non-disease group.Rabbits in control group were similarly injected with the same volume of 9 g/L saline.Penicillin sodium 50 000 U/rabbit was injected once a week for preventing infection.After 8 weeks of injection, CT was performed in all the experimental animals.They were then sacrificed for collecting bilateral femoral heads.Expression levels of TGF-β1, TGF-β2, Smad2 and Smad3 in the femoral head were detected by enzyme linked immunosorbent assay (ELISA), and the mRNA level of Runx2 in the femoral head was detected by quantitative real-time PCR (qPCR), the expression differences of related factors in each group were compared.Results:In steroid injection group (48 cases), 6 rabbits were sacrificed, and 32 survived, involving 6/32 cases (18.75%) experimental animals with positive avascular necrosis (disease group), and 26 negative ones (non-disease group). ELISA data showed that expression levels of TGF-β1 in control group, non-disease group and disease group were (77.12±14.62) ng/L, (90.17±11.90) ng/L and (126.14±25.66) ng/L, respectively ( t=3.35, 4.24, all P<0.05). The expression levels of TGF-β2 in control group, non-disease group and disease group were (74.54±7.63) ng/L, (89.24±9.51) ng/L and (109.74±16.45) ng/L, respectively ( t=4.12, 5.65, all P<0.01). The expression levels of Smad2 in control group, non-disease group and disease group were (17.74±2.72) μg/L, (23.82±3.58) μg/L and (31.28±3.88) μg/L, respectively ( t= 4.54, 7.99, all P<0.01). The expression levels of Smad3 in control group, non-disease group and disease group were (1.76±0.52) μg/L, (2.39±0.45) μg/L and (3.53±0.47) μg/L, respectively ( t=5.60, 6.71, all P<0.01). qPCR data showed that the mRNA levels of Runx2 in control group, non-disease group and disease group were 1.02±0.17, 1.27±0.14, and 1.72±0.11, respectively ( t=7.60, 8.91, all P<0.01). Conclusions:TGF-β is up-regulated in the model of steroid-induced osteonecrosis of the femoral head in young rabbits, which stimulates the proliferation and differentiation of osteoblasts and osteoclasts, and triggers the process of bone remodeling.The TGF-β signaling pathway involved in the repair of necrotic bone.

Objective To investigate the cell origin of interleukin (IL)-22-secreting cell of mice infected with Trichinella spiralis (T.spiralis) at the early encapsulated stage.Methods Twelve Balb/c mice were divided into the infected group and the control group according to body weight by random number table.The infected mice were intragastrically administrated with 300 muscle larvae of T.spiralis,and the control mice were given the same amount of normal saline.The IL-22-secreting cell subsets in mouse splenic lymphocytes were detected by flow cytometry at the fourth week after infection.Results The proportion of IL-22-secreting cells in splenic lymphocytes of T.spiralis infected mice was increased when compared with control group [(0.88 ± 0.25)％ vs (0.28 ±0.17)％,t =-4.899,P ＜ 0.05].There was no significant difference between the proportion of CD3+IL-22+ cells and CD3-IL-22+ cells in the splenic lymphocytes of the infected group [(0.29 ± 0.17)％ vs (0.51 ± 0.17)％,t =-2.195,P ＞ 0.05],and the percentage of CD3-IL-22+ cells were similar between the infected group and the control group [(0.51 ± 0.17)％ vs (0.44 ± 0.22)％,t =-0.600,P ＞ 0.05].The proportion of CD3+IL-22+ cells in the infected group was significantly higher than that in the control group [(0.29 ± 0.17)％ vs (0.07 ± 0.06)％,t =-3.068,P ＜ 0.05],and the percentage of CD4+IL-22+ T cells and γδTCR+IL-22+ T cells were obviously increased in CD3+ lymphocytes [(1.28 ± 0.54)％ vs (0.16 ± 0.07)％,(0.33 ± 0.22)％ vs (0.02 ± 0.00)％,t =-4.997,-3.342,P ＜ 0.05].Conclusions The proportion of IL-22-secreting splenic lymphocytes is increased in mice infected with T.spiralis at the early encapsulated stage.The rise is caused by increased numbers of IL-22-secreting CD3 + lymphocytes,especially CD4+ T cells and γδT cells.