Objective:To explore the effect of mesoporous silica (SiO 2) chitosan (CS) hydrogel loaded with neurotrophin-3 (NT-3) on repair of spinal cord injury. Methods:After 2%, 4%, and 6% NT-3/SiO 2 were dissolved in CS solution, they were added into β-glycerophosphate sodium solution for chemical cross-linking to obtain hydrogel patches of different NT-3 loadings. The specific surface area, pore size and pore volume of mesoporous SiO 2 nanoparticles were detected by specific surface area analyzer. The morphology of mesoporous SiO 2 nanoparticles and the pore structure of freeze-dried hydrogel were detected by scanning electron microscopy. Adhesion of the hydrogel was verified by spinal cord tissue. After the NT-3/SiO 2@CS hydrogel was placed in the medium, the concentrations of NT-3 were measured for 1 to 20 days. Neural stem cells (NSCs) were isolated from fetal rats and identified. Cell counting kit 8 (CCK-8) assay was used to detect the proliferation of NSCs treated with different concentrations of hydrogel. Immunofluorescence staining was used to detect the effects of NT-3/SiO 2@CS hydrogel on differentiation of NSCs. Twenty-four 8-week-old C57BL/6JNifdc female mice were randomly divided into a sham operation group (sham), a spinal cord injury group (SCI), a chitosan hydrogel group (CS) and a mesoporous SiO 2-loaded NT-3 hydrogel group (NT-3/SiO 2@CS). In the sham group, the muscle and skin were sutured immediately after laminectomy without spinal cord injury. The CS hydrogel and NT-3/SiO 2@CS hydrogel patches were implanted without treatment after spinal cord injury in the other 3 groups, respectively. Basso Mouse Scale (BMS) was used to evaluate the mice every 7 days for 8 weeks after modeling. The hot and cold board test and Catwalk gait analysis were performed at 8 weeks after surgery. Results:The mesoporous SiO 2 nanoparticles showed typical spherical morphology and a uniform particle size (about 160 nm). The specific surface area, pore volume and pore size of the mesoporous SiO 2 nanoparticles loaded with NT-3 changed from 1,039 m 2/g, 0.726 cm 3/g and 2.754 nm to 779 m 2/g, 0.403 cm 3/g and 1.903 nm, respectively. The hydrogel had a uniform internal microporous structure and good porosity so that it easily adhered to the spinal cord and achieved long-term stable release of NT-3 for at least 20 days. CCK-8 results showed that at 24 hours after the neural stem cells were laid, the cell proliferative activities in the 4%NT-3/SiO 2@CS and 6% NT-3/SiO 2@CS groups were significantly lower than that in the group untreated ( P<0.05). The immunofluorescence staining showed that the fluorescence intensity of glial fibrillary acidic protein (GFAP), the NSC marker in NT-3/SiO 2@CS, insignificantly decreased in the groups with different concentrations of NT-3/SiO 2@CS compared with the group untreated ( P>0.05). The fluorescence intensities of GFAP, MAP2 and GFAP/MAP2 in the NT-3/SiO 2@CS group were significantly higher than those in the other groups ( P<0.05). The BMS scoring for mice showed sham group>NT-3/SiO 2@CS group>SCI group and CS group for 8 weeks of modeling, and the response time of mice to cold and hot stimulations in the NT-3/SiO 2@CS group was significantly shorter than that in the SCI and CS groups. The differences above were statistically significant ( P<0.05). The Catwalk gait analysis showed that the hindlimb footprints in the NT-3/SiO 2@CS group were significantly clearer and more coherent than those in the SCI and CS groups. Conclusions:The sustained-release gel patch based on CS, SiO 2 and NT-3 has a uniform pore structure, good biocompatibility and excellent drug sustained-release effect. It can promote the differentiation of NSCs into neurons, thus contributing to recovery of motor and sensory functions after spinal cord injury.

Objective:To compare the performance differences of domestic and imported CD3/CD28 activation beads for manufacturing CAR-T cells,providing a backup or alternative for domestic CAR-T cell research and manufacture.Methods:A mature protocol using imported CD3/CD28 activation beads with a 1∶1 ratio for CD3+T cells was implemented as research control.Domestic beads were used with gradient ratios of 1∶2,1∶1,and 2∶1 to activate T cells.72 h after T cell activation,CAR-T cells were manufactured by CAR lentiviral infection and cell proliferation was monitored at 2-,4-,and 7-days post-infection.Flow Cytometry was used to detect CAR-T cell positivity 5 days after infection and to detectCD4/CD8 phenotype of CAR-T cells and PD1+TIM3+cell exhaustion ratio 8 days after infection.Results:CAR-T cells manufactured by domestic CD3/CD28 activation beads exhibited similar phenotype compared with those manufactured by imported CD4/CD8 beads.The positive rate of CAR-T cells prepared with domestic beads was slightly lower than that of imported beads(53.7%vs 57.9%).However,the proliferation of CAR-T cells manufactured by domestic beads was about twice that of cells manufactured by imported beads,and the exhaustion level was only half that of imported beads(4.21%vs 7.91%).Moreover,the use of domestic magnetic beads was lower than that of imported magnetic beads,which was advantageous for cutting the costs of CAR-T cells research and manufacture.Conclusion:Domestic CD3/CD28 activation beads used for CAR-T cells manufacturing demonstrate comparable overall performance to their imported counterparts,showing potential as a backup or alternative for imported beads.

Objective:To explore the effect of mesoporous silica (SiO 2) chitosan (CS) hydrogel loaded with neurotrophin-3 (NT-3) on repair of spinal cord injury. Methods:After 2%, 4%, and 6% NT-3/SiO 2 were dissolved in CS solution, they were added into β-glycerophosphate sodium solution for chemical cross-linking to obtain hydrogel patches of different NT-3 loadings. The specific surface area, pore size and pore volume of mesoporous SiO 2 nanoparticles were detected by specific surface area analyzer. The morphology of mesoporous SiO 2 nanoparticles and the pore structure of freeze-dried hydrogel were detected by scanning electron microscopy. Adhesion of the hydrogel was verified by spinal cord tissue. After the NT-3/SiO 2@CS hydrogel was placed in the medium, the concentrations of NT-3 were measured for 1 to 20 days. Neural stem cells (NSCs) were isolated from fetal rats and identified. Cell counting kit 8 (CCK-8) assay was used to detect the proliferation of NSCs treated with different concentrations of hydrogel. Immunofluorescence staining was used to detect the effects of NT-3/SiO 2@CS hydrogel on differentiation of NSCs. Twenty-four 8-week-old C57BL/6JNifdc female mice were randomly divided into a sham operation group (sham), a spinal cord injury group (SCI), a chitosan hydrogel group (CS) and a mesoporous SiO 2-loaded NT-3 hydrogel group (NT-3/SiO 2@CS). In the sham group, the muscle and skin were sutured immediately after laminectomy without spinal cord injury. The CS hydrogel and NT-3/SiO 2@CS hydrogel patches were implanted without treatment after spinal cord injury in the other 3 groups, respectively. Basso Mouse Scale (BMS) was used to evaluate the mice every 7 days for 8 weeks after modeling. The hot and cold board test and Catwalk gait analysis were performed at 8 weeks after surgery. Results:The mesoporous SiO 2 nanoparticles showed typical spherical morphology and a uniform particle size (about 160 nm). The specific surface area, pore volume and pore size of the mesoporous SiO 2 nanoparticles loaded with NT-3 changed from 1,039 m 2/g, 0.726 cm 3/g and 2.754 nm to 779 m 2/g, 0.403 cm 3/g and 1.903 nm, respectively. The hydrogel had a uniform internal microporous structure and good porosity so that it easily adhered to the spinal cord and achieved long-term stable release of NT-3 for at least 20 days. CCK-8 results showed that at 24 hours after the neural stem cells were laid, the cell proliferative activities in the 4%NT-3/SiO 2@CS and 6% NT-3/SiO 2@CS groups were significantly lower than that in the group untreated ( P<0.05). The immunofluorescence staining showed that the fluorescence intensity of glial fibrillary acidic protein (GFAP), the NSC marker in NT-3/SiO 2@CS, insignificantly decreased in the groups with different concentrations of NT-3/SiO 2@CS compared with the group untreated ( P>0.05). The fluorescence intensities of GFAP, MAP2 and GFAP/MAP2 in the NT-3/SiO 2@CS group were significantly higher than those in the other groups ( P<0.05). The BMS scoring for mice showed sham group>NT-3/SiO 2@CS group>SCI group and CS group for 8 weeks of modeling, and the response time of mice to cold and hot stimulations in the NT-3/SiO 2@CS group was significantly shorter than that in the SCI and CS groups. The differences above were statistically significant ( P<0.05). The Catwalk gait analysis showed that the hindlimb footprints in the NT-3/SiO 2@CS group were significantly clearer and more coherent than those in the SCI and CS groups. Conclusions:The sustained-release gel patch based on CS, SiO 2 and NT-3 has a uniform pore structure, good biocompatibility and excellent drug sustained-release effect. It can promote the differentiation of NSCs into neurons, thus contributing to recovery of motor and sensory functions after spinal cord injury.

Objective:To compare the diagnostic efficacy of 22 G fine needle aspiration (FNA) needles and 22 G end-cutting fine needle biopsy (FNB) needles for solid pancreatic lesion using both cytological and histological examination.Methods:Clinical data of 116 patients who underwent endoscopic ultrasound-guided fine needle aspiration/biopsy (EUS-FNA/FNB) at the Digestive Endoscopy Center of Ruijin Hospital, Shanghai Jiao Tong University School of Medicine from June 2022 to March 2023 were retrospectively analyzed. Sixty-three patients sampled with 22 G FNA needles were the FNA group, and 53 sampled with 22 G FNB needles were the FNB group. The diagnostic accuracy, sensitivity, specificity, positive predictive value, negative predictive value, and cytological and histological diagnostic yield of FNA needles and FNB needles for solid pancreatic lesions were compared.Results:There were no significant differences in age, gender, lesion location, lesion size, or the number of passes between the FNA group and the FNB group ( P>0.05). There were no significant differences in the diagnostic accuracy [93.7% (59/63) VS 90.6% (48/53), P=0.730], sensitivity [93.0% (53/57) VS 90.2% (46/51), P=0.732], specificity [100.0% (6/6) VS 100.0% (2/2), P=1.000], positive predictive value [100.0% (53/53) VS 100.0% (46/46), P=1.000] and negative predictive value [60.0% (6/10) VS 28.6% (2/7), P=0.335] of combined cytology and histology in distinguishing benign and malignant lesions between the two groups. In the FNA group, the diagnostic accuracy of combined cytology and histology was higher than cytology alone [93.7% (59/63) VS 81.0% (51/63), P=0.008], and was higher than histology alone without statistical significance [93.7% (59/63) VS 87.3% (55/63), P=0.125]. In the FNB group, the diagnostic accuracy of combined cytology and histology was higher than cytology alone [90.6% (48/53) VS 69.8% (37/53), P=0.001], but not than histology alone [90.6% (48/53) VS 90.6% (48/53)， P=1.000]. For solid masses located in pancreatic body/tail, the diagnostic accuracy for malignancy by histology using FNB needles tended to be higher than that of FNA needles [100.0% (17/17) VS 81.3% (26/32), P=0.080]. Conclusion:Both FNA needles and FNB needles exhibit adequate diagnostic yield for solid pancreatic masses when combining cytology and histology. FNB needles may offer a higher histological diagnostic yield.

Objective:To investigate the clinicopathologic features and prognosis of mediastinal T lymphoblastic lymphoma/leukemia (T-LBL/ALL).Methods:Sixty-one patients with mediastinal T-LBL/ALL diagnosed at First Affiliated Hospital of Zhengzhou University from August 1, 2011 to December 31, 2018 were enrolled. Their clinical, pathological, imaging features and prognosis were retrospectively analyzed.Results:Of the 61 patients with mediastinal T-LBL/ALL, 46 were male and 15 were female, with a male to female ratio of approximately 3∶1, aged 5 to 71 years (median 24 years, average of 24.5 years). Radiological findings were mediastinal soft tissue masses (58 cases) or mediastinal multiple enlarged lymph nodes (1 case). The tumor had a diameter of 4.9 to 18.3?cm in size, and data of 2 cases was unavailable. The patient′s main symptoms were superior vena cava syndrome (cough, dyspnea, facial or neck edema), shortness of breath and chest pain, while about 1/3 of patients developed B symptoms (high fever, night sweats or significant weight loss). All 61 cases were biopsy specimens, and 2 of the tumors were later resected. Histopathologic examination showed that the thymic tissue epithelial network structure was destroyed or completely disappeared. A large number of lymphocytoid tumor cells were diffusely infiltrative, with infiltration into adipose tissue, starry sky phenomenon, linear-like arrangement, interstitial collagen hyperplasia and tumor cell extrusion. Focal tumor necrosis was present in some cases. Tumor cells were overall small to medium in size. They had little cytoplasm, slightly distorted, round or oval-shaped nuclei, fine chromatin, and innocuous/small nucleoli. Immunohistochemical studies showed that the tumor cells expressed CD7 (100%, 33/33), TDT (93.4%, 57/61), CD99 (83.3%, 25/30), CD1a (4/7), CD10 (8/18), CD34 (13.2%, 5/38), but did not express B cell markers (CD20 and PAX5) or granulocyte monocyte marker (MPO). The Ki-67 proliferation index was usually greater than 50%. One case was tested for TCR clonal rearrangement, which was positive. Several hemotherapy regiments were used. Hyper-CVAD (cyclophosphamide, vindesine, dexamethasone, and epirubicin) were most frequently administrated (60.4%, 32/53), followed by BFM-90 (50.9%, 27/53). Some patients were treated with the above two and other treatment options. Follow-up data were available in 55 of the 61 patients, and 26 patients (47.3%) survived. The average five-year survival rate was 50.6%. The patient′s prognosis was not significantly related to the International Prognostic Index, age of onset, gender, or tumor size.Conclusions:The mediastinal T-LBL/ALL is rare, and most of its specimens are needle biopsies. The histological morphology is often difficult to interpret, while the addition of clinical features and immunohistochemistry may help. The combination of CKpan, TDT, CD99, CD7, CD3, PAX5, CD34, CD10, and Ki-67 immunohistochemicl studies may assist in diagnosis of the most cases.

Objective:To investigate the clinicopathological features of pulmonary epithelioid hemangioendothelioma (PEHE).Methods:Eighteen cases of PEHE were collected from August 2011 to December 2018 at the First Affiliated Hospital of Zhengzhou University. All cases were retrospectively studied by hematoxylin and eosin staining and immunohistochemistry (IHC). The clinicopathological features were reviewed; the status of CAMTA1 and TFE3 gene was analyzed and patients′ outcome was followed up.Results:Of the 18 cases, there were 11 males and 7 females with a male to female ratio of 1.6 to 1.0. The patients′ age ranged from 36 to 68 years (mean 52 years). Twelve cases (12/18) showed a single nodule and six cases (6/18) showed multiple bilateral nodules. Seven cases (7/18) involved other organs besides lung. Seventeen (17/18) patients presented with respiratory symptoms and one patient (1/18) presented with abdominal pain. Grossly, the tumors were greyish-white nodules with indistinct borders. Microscopically the tumor cells were epithelioid and arranged in strands and nests, and cytoplasmic vacuoles were commonly noted. The stroma was myxochondroid or hyaline. By IHC, the tumor cells were positive for CD31(18/18), CD34 (16/18), ERG (18/18) and Fli-1 (18/18); CKpan was focally positive in 5 cases (5/18). TFE3 was positive in 3 cases (3/18), and Ki-67 index ranged from 5% to 30%. FISH analysis showed seventeen cases (17/18) had CAMAT1 rearrangement, one case had TFE3 rearrangement displaying a split signal. Eight patients (8/18) had surgical excision, three patients (3/18) had surgery and chemotherapy, and seven patients (7/18) had chemotherapy only. Four patients (4/18) died of the disease.Conclusions:Patients with PEHE have non-specific symptoms, and correct diagnosis depends on pathologic biopsy and the exclusion of other tumors with epithelioid morphology. Some patients with PEHE have poor prognosis, particularly in those who have multiple nodules, peripheral invasion or metastasis.

Objective:To investigate the expression of microRNA-140-5p (miR-140-5p) in esophageal squamous cell carcinoma (ESCC) and its role in cell proliferation and invasion of ESCC.Methods:Real-time quantitative PCR (qPCR) was used to detect the expression levels of miR-140-5p in ESCC tissues and cells. Negative control and miR-140-5p mimic were transfected into Eca109 and KYSE70 cells. CCK-8 kit and Transwell assay were employed to examine the changes of cell proliferation and invasion ability after transfection, respectively. The dual-luciferase reporter assay was used to assess the interaction of miR-140-5p with Glut1. Western blot was utilized to detect the Glut1 protein expression after transfection.Results:Analysis of the related GEO datasets revealed that the expression of miR-140-5p in ESCC tissues was significantly lower than that in normal tissues ( P<0.01). The qPCR testing demonstrated that the expression of miR-140-5p in ESCC tissues and cells was markedly lower than that in normal tissues and normal esophageal epithelial cell Het-1A ( P<0.01). The miR-140-5p expression was closely associated with tumor differentiation, TNM staging and lymph node metastasis in ESCC patients. The survival rate of ESCC patients with high miR-140-5p level was higher than those with low miR-140-5p level ( P<0.05). Besides, addition of miR-140-5p mimic significantly upregulated the expression of miR-140-5p in Eca109 and KYSE70 cells, and suppressed cell proliferation and invasion in Eca109 and KYSE70 cells. The dual-luciferase reporter assay showed that Glut1 was a direct target of miR-140-5p in ESCC cells, and its expression was upregulated in ESCC tissues. Glut1 expression was inversely associated with miR-140-5p expression in ESCC tissues. MiR-140-5p mimic dramatically inhibited the expression of Glut1 in Eca109 and KYSE70 cells. Conclusions:MiR-140-5p plays an essential role in ESCC development and progression. Targeting at miR-140-5p/Glut1 may be a novel therapeutic strategy for ESCC patients.

Objective: To compare the clinical effects between minimally invasive transforaminal lumbar interbody fusion (MIS-TLIF) assisted by Microendoscopic discectomy (MED) and Quadrant for the treatment of degenerative lumbar spinal stenosis (DLSS). Methods: All of 59 patients suffered from DLSS treated surgically from May 2015 to October 2017 were reviewed. According to the surgery method, all cases were divided into MED group (27 cases) and Quadrant channel group (32 cases). All patients were followed up for an average of 18.5 months (11-29 months). Comparison was made on the operative time, intraoperative blood loss, postoperative drainage, postoperative time in bed, postoperative creatine kinase (CK), fusion rate and the degree of muscle fibrosis shown in MRI, as well as visual analogue scale (VAS)score and Oswestry dysfunction index (ODI) score in two groups. Results: The duration of operation in MED group was significantly longer than that in Quadrant group (161.7±22.4 min vs. 145.6±19.4 min, t=4.541, P<0. 01), but less intraoperative blood loss (138.1±26.9 ml vs. 155.6±21.5 ml, t=-2.724, P< 0. 01) and shorter time in bed after surgery (2.3±0.7 d vs. 3.5±1.1 d, t=-4.564, P<0.01). Compared with Quadrant group, CK levels were risen slightly on the first and third postoperative day (P<0.01). CK on the fifth postoperative day in both groups returned to normal. VAS score of lower back pain in MED group was lower than that in Quadrant group on the third day and twelfth month after operation (P<0.05). After 3 months, there was no significant difference in VAS score and ODI between the two groups (P>0.05). ODI was lower in MED group than that in Quadrant group after 6 months and 12 months.The fusion rate was88.9%(24/27) in MED group and 93.8%(30/32) in Quadrant channel group. There was no statistical difference in fusion rate of two groups. 10 patients in MED group and 12 patients in Quadrant group underwent MRI examination of lumbar spine one year after operation. The ratio of postoperative and preoperative atrophy of multiplex muscle area was measured. Muscle atrophy of lower back muscle was lighter in MED group (0.12±0.05 vs. 0.22±0.04, t=-2.428, P<0.05). For intraoperative and postoperative complications, 1 case of dural sac rupture occurred in both groups. Gelatin sponge immediately with fibrin glue was used for plugging up, no postoperative cerebrospinal fluid leakage was found. In Quadrant channel group, 1 case had less blood supply of skin incision edges and epidermal necrosis while the other case had fat liquefaction. Conclusion Compared with the aid of Quadrant, MIS-TLIF assisted with MED had less blood loss, less trauma and faster recovery and could reduce the incidence of postoperative incision complication.

Objective: To investigate the clinicopathological features, diagnosis and differential diagnosis of pulmonary extranodal NK/T-cell lymphoma, nasal type (ENKTL-NT). Methods: Eleven cases of newly diagnosed (10 puncture biopsies and 1 transbronchial biopsy), previously untreated pulmonary ENKTL-NT were collected at the First Affiliated Hospital of Zhengzhou University, from August 2013 to November 2018. The clinicopathological features including histomorphology, immunohistochemistry and in situ hybridization were collected and analyzed. Results: Among the 11 cases, 8 were males and 3 were females, with a male to female ratio of 8∶3.The age range was from 30 to 74 years, with an average of 48 years and a median of 43 years. Tumors involved bilateral lung lobes in 8 cases, the upper left lobe in 1 case, lower left lobe in 1 case, and upper right lobe in 1 case. Main clinical symptoms included fever, often accompanied by cough, and bloody sputum in most cases. All cases were stage Ⅳ E. Histological features included scattered or focal aggregates of marked pleomorphic tumor lymphocytes, accompanied by necrosis and heavy admixture of inflammatory cells. In a few cases, diffuse neoplastic lymphocytes or vascular central and destructive infiltrations were seen. Tumor cells in most cases expressed CD3ε, CD3, CD43, CD56, TIA-1, granzyme B, but did not express CD20, CD79a, and CD5. Ki-67 index ranged from 40%to 90%.All cases were positive for EBER by in situ hybridization. Four of five patients died during follow-up with a survival period of only 1 week to 13 months. Conclusions Pulmonary ENKTL-NT is rare, high grade malignancy with a poor prognosis. Misdiagnosis is common due to lesional necrosis and heterogeneous cell components. Immunohistochemistry and EBER in situ hybridization are essential for accurate diagnosis.

Objective: To investigate the clinicopathological features and prognostic indicators of primary pulmonary adenoid cystic carcinoma. Methods: Fifty-nine cases of primary pulmonary adenoid cystic carcinoma were collected from August 2011 to December 2017 at the First Affiliated Hospital of Zhengzhou University. All cases were retrospectively studied by hematoxylin-eosin staining and immunohistochemistry. The clinicopathological features were reviewed and patient survival analysis was performed using Kaplan-Meier method and Cox regression model. Status of epidermal growth factor receptor (EGFR), KRAS, BRAF genes was analyzed in 15 of the 59 study cases. Results: Among 59 cases, there were 25 males and 34 females with male to female ratio of 1.0 to 1.4. The patient age ranged from 29 to 81 years with a mean age of 55 years. The tumor max diameters ranged from 1.0 to 9.6 cm with an average diameter of 2.8 cm. Fifteen (25.4%) patients were smokers while 44 patients (74.6%) were non-smokers. Tumors predominantly occurred in the trachea (28/59,47.5%), the left main bronchus (7/59,11.9%) and the right bronchus (5/59,8.5%). Grossly, the tumors were well circumscribed, greyish-white nodules. Microscopically the tumor cells were small and uniform, and arranged in tubular, cribriform, and solid patterns. Immunohistochemistry showed that the tumor cells were positive for CK7, S-100 protein, Sox-10, CD117 and p63. TTF1 was only positive in 2 cases and Ki-67 index ranged from 3% to 40%. Eighteen cases (30.5%) were gradeⅠ, 26 cases (40.1%) grade Ⅱ, and 15 cases (25.4%) grade Ⅲ. Overall, 39 cases (66.1%), 7 cases (11.9%), 10 cases (16.9%), and 3 cases (5.1%) were at stages Ⅰ, Ⅱ, Ⅲ, and Ⅳ, respectively. Twenty-three patients (39.0%) received surgical therapy, 3 patients (5.1%) surgery combined with radiotherapy, 9 patients (15.2%) surgery combined with chemotherapy, and 24 cases (40.7%) chemotherapy only. No mutation of EGFR, KRAS and BRAF was detected in all 15 tested cases. The overall survival rate at the first, third and fifth years was 94.9%, 86.4% and 84.7%, respectively. Prognostic analysis showed that patient′s age and tumor size were statistically associated with the survival (P<0.05). Conclusions Majority of the patients with primary pulmonary adenoid cystic carcinoma are at an early clinical stage with a favorable prognosis. The size of the tumor and the age of the patients are independent prognostic indicators.