Umbilical cord mesenchymal stem cells (UC-MSCs) have been widely used in regenerative medicine, but there is limited research on the stability of UC-MSCs formulation during production. This study aims to assess the stability of the cell stock solution and intermediate product throughout the production process, as well as the final product following reconstitution, in order to offer guidance for the manufacturing process and serve as a reference for formulation reconstitution methods. Three batches of cell formulation were produced and stored under low temperature (2-8 ℃) and room temperature (20-26 ℃) during cell stock solution and intermediate product stages. The storage time intervals for cell stock solution were 0, 2, 4, and 6 h, while for intermediate products, the intervals were 0, 1, 2, and 3 h. The evaluation items included visual inspection, viable cell concentration, cell viability, cell surface markers, lymphocyte proliferation inhibition rate, and sterility. Additionally, dilution and culture stability studies were performed after reconstitution of the cell product. The reconstitution diluents included 0.9% sodium chloride injection, 0.9% sodium chloride injection + 1% human serum albumin, and 0.9% sodium chloride injection + 2% human serum albumin, with dilution ratios of 10-fold and 40-fold. The storage time intervals after dilution were 0, 1, 2, 3, and 4 h. The reconstitution culture media included DMEM medium, DMEM + 2% platelet lysate, 0.9% sodium chloride injection, and 0.9% sodium chloride injection + 1% human serum albumin, and the culture duration was 24 h. The evaluation items were viable cell concentration and cell viability. The results showed that the cell stock solution remained stable for up to 6 h under both low temperature (2-8 ℃) and room temperature (20-26 ℃) conditions, while the intermediate product remained stable for up to 3 h under the same conditions. After formulation reconstitution, using sodium chloride injection diluted with 1% or 2% human serum albumin maintained a viability of over 80% within 4 h. It was observed that different dilution factors had an impact on cell viability. After formulation reconstitution, cultivation in medium with 2% platelet lysate resulted in a cell viability of over 80% after 24 h. In conclusion, the stability of cell stock solution within 6 h and intermediate product within 3 h meets the requirements. The addition of 1% or 2% human serum albumin in the reconstitution diluent can better protect the post-reconstitution cell viability.

Objective:To develop a nomogram predictive model on the basis of identification of the risk factors associated with failure in repositioning the dislocation of the subaxial cervical spine with locked facets by skull traction.Methods:A retrospective study was conducted of the clinical data of the patients who had been treated for dislocation of the subaxial cervical spine with locked facets at Department of Orthopaedic Trauma, The Hospital Affiliated to Guizhou Medical University and Department of Spine Surgery, The People's Hospital of Guizhou Province from January 2014 to December 2022. The clinical data from The Hospital Affiliated to Guizhou Medical University were used as a training set (156 cases) and those from The People's Hospital of Guizhou Province as an external validation set (54 cases). Univariate analysis and multi-variate logistic regression analysis of the training set were conducted to screen out independent risk factors associated with the failure in repositioning the dislocation of the subaxial cervical spine with locked facets by skull traction. A nomogram predictive model was thus constructed and assessed by the receiver operating characteristic (ROC) curve, calibration curve, and decision curve. Internal validation of the training set and external validation set was used to evaluate and validate the model.Results:The multivariate logistic regression analysis revealed that cervical Ⅰ grade dislocation ( P=0.002), cervical Ⅱ grade dislocation ( P=0.007), low segment affected ( P=0.042), unilateral facet locked ( P=0.027), and the ASIA grading of spinal cord injury ( P=0.008) were the independent risk factors associated with the failure in repositioning the dislocation of the subaxial cervical spine with locked facets by skull traction, based on which the nomogram model with a C-index of 0.88 was constructed to predict the failure in repositioning the dislocation of the subaxial cervical spine with locked facets by skull traction. Analysis of the ROC curve of the training set showed an area under the curve (AUC) of 0.88, indicating good accuracy of the nomogram model. Analysis of the calibration curve showed high consistency between the probability of the nomogram model predicting the failure in repositioning the dislocation of the subaxial cervical spine with locked facets by skull traction and the actual probability of traction reposition failure. Analysis of the decision curve showed that application of the nomogram model led to good benefits when the net benefit threshold for the failure in repositioning the dislocation of the subaxial cervical spine with locked facets by skull traction was 0.03 to 0.84. Analysis of the ROC curve of external validation set showed an AUC of 0.79, indicating good accuracy of the nomogram model. The training set showed a C-index of 0.87 after 1,000 internal verifications by the Bootstrap method, indicating good discrimination of the nomogram model. Conclusions:Cervical Ⅰ grade dislocation, cervical Ⅱ grade dislocation, low segment affected, unilateral facet locked, and incomplete spinal cord injury are independent risk factors associated with failure in repositioning the dislocation of the subaxial cervical spine with locked facets by skull traction. A nomogram model has been successfully constructed which can predict the failure in repositioning the dislocation of the subaxial cervical spine with locked facets by skull traction. Validation and evaluation of the nomogram model have demonstrated its good predictive value.

【Objective】 To investigate the effects of silencing AEG-1 gene by shRNA on vasculogenic mimicry (VM) of a glioma xenograft model. 【Methods】 U87 glioma cells were infected with AEG-1 shRNA lentivirals. Real-time PCR and Western blotting were used to detect the mRNA and protein expressions of AEG-1 in U87 cells after infected by the AEG-1 shRNA lentivirals. A glioma xenograft model was generated and CD 34/PAS double-staining was performed to detect the VM channels in vivo. The immunohistochemical assay was performed to evaluate the expressions of MMP-2, MMP-9, VE-cadherin, and VEGF in glioma xenograft models. 【Results】 AEG-1 shRNA lentivirals could significantly inhibit the AEG-1 expression in glioma cells (P<0.01). Meanwhile, they also decreased the number of VM in the glioma xenograft model (P<0.01). Furthermore, the expressions of MMP-2, MMP-9, VE-cadherin, and VEGF in glioma significantly decreased in vivo (P<0.01). 【Conclusion】 These results suggest that silencing AEG-1 gene by shRNA can significantly inhibit VM of glioma in vivo, the mechanism of which may partly be through regulating MMP-2, MMP-9, VEGF, and VE-cadherin expressions.

Objective To explore the effect of 2-chloroethyl ethyl sulfide(CEES) poisoning on keratinocyte migration and the regulatory role of microRNA(miR)-34a.Methods MTS was used to detect the viability of cells exposed to CEES in order to select an appropriate dose of CEES exposure in this in vitro model.The protein level of keratin 5 and keratin 10 was detected to assess cell differentiation status .Scratch assay was applied to evaluate cell migration ,and miR-34a silencing in keratinocytes was achieved by transfecting chemically synthesized miR-34a specific miRNA inhibitor.t-ERK1/2 and p-ERK1/2 levels closely related to cell migration were detected using Western blotting .Results An in vitro CEES exposure model of keratinocytes was established at the optimal concentration of 0.5 mmol/L CEES in the viability test , and this dose was chosen to evaluate cell migration changes .The migration of cells was significantly inhibited 24 h after CEES exposure , accompanied by no changes in morphology and keratin 5/10 levels.Silencing of miR-34a significantly increased the migration of cells exposed to CEES , which could be blocked by adding 5 μmol/L U0126 , an ERK1/2 phosphorylation selective inhibitor.Conclusion Silencing of miR-34a can significantly increase keratinocyte migration and partially reverse the inhibition of CEES-caused migration , which could be mediated by ERK 1/2 pathway activation .

Objective To master the level of iodine nutrition among population in Jiangxi province,and to provide a scientific basis for establishing the strategy for prevention and control of iodine deficiency disorders (IDD).Methods Retrospective method was adopted to analyze the goiter rate and frequency distribution of urinary iodine of children aged 8-10,the qualified rate of iodized salt,the coverage rate of iodized salt and the consumption rate of qualified iodized salt in residents of Jiangxi province from 1995 to 2010.The method of correlation analysis was used to analyze the relationship between goiter rate of children (by palpation) and the qualified rate of iodized salt,iodized salt coverage rate and residents consumption rate of qualified iodized salt.Results The goiter rates (measured by the method of palpation) of children aged 8-10 were down from 40.17％(482/1200) in 1995 to 0.80％(16/2000) in 2010(x2 =4.864,P＜ 0.05).The median of urinary iodine of children was higher than 200 μg/L; the proportion of people whose urinary iodine content higher than 300 μg/L was above 25.00％ and the highest propoaion was up to 58.01％ (210/362) between 1995-2010.The minimum median of salt iodine was 17.77 mg/kg in 1995,and 29.30-39.10 mg/kg in other years.The qualified rates of iodized salt,the iodized salt coverage rates and the consumption rates of qualified iodized salt increased from 43.58％(452/1037),86.42％(1037/1200) and 37.67％(452/1200) in 1995 to 97.95％ (1916/1956),99.95％(1956/1957) and 97.90％(1916/1957) in 2010,respectively; there was a growth trend over the years(x2 =5.240,6.118,5.631,all P ＜ 0.05).The goiter rates of children were related to the qualified rates of iodized salt,the iodized salt coverage rates and the consumption rates of qualified iodized salt,and the correlation coefficient(r) was-0.833,-0.881 and-0.918 (all P ＜ 0.05),respectively.Conclusions The level of iodine nutrition among residents in Jiangxi province has already gone beyond the appropriate level,and the iodine concentration in salt should be cut to ensure the appropriate iodine nutrition level among people.