BACKGROUND: Previous studies have shown that neurotrophin 3 (NT3)-chitosan can induce endogenous neurogenesis and axon regeneration in rats with spinal cord injury, and promote recovery of motor and sensory functions in rats. OBJECTIVE: To observe the effect of rehabilitation training combined with NT3-chitosan biomaterial scaffold on skeletal muscle morphological changes and functional recovery in rats with complete spinal cord injury. METHODS: Fifty adult female Wistar rats were randomly divided into five groups, 10 in each group. The sham group was not modeled; the remaining four groups were prepared with T7-T8 complete 5-mm spinal cord injury model, and the lesion control was not performed any intervention after modeling. The other three groups were given rehabilitation training, NT3-chitosan active biomaterial scaffold, NT3-chitosan active biomaterial scaffold combined with rehabilitation training intervention. Rehabilitation training started 2 weeks after modeling. Before operation, 2, 4, 6, 8, 10, and 12 weeks after operation, all of rats were subjected to double-blind open-field BBB scores. After 12 weeks, the skeletal muscles of the hind limbs (tibialis anterior muscle, gastrocnemius muscle, and soleus muscle) were taken for hematoxylin-eosin staining and acetylcholinesterase staining. The changes in muscle atrophy and motor endplates were assessed in each group. The experimental plan was approved by the Animal Experiment Committee of Capital Medical University (approval No. AEEI-2018-105). RESULTS AND CONCLUSION: (1) The BBB score at each time point in the sham group was higher than that in the other four groups (P < 0.05); and the scores at 8, 10, and 12 weeks after the NT3-chitosan combined rehabilitation training group were higher than the lesion control group, the lesion control combined rehabilitation training group, and NT3-chitosan group (P < 0.05). (2) At 12 weeks after operation, hematoxylin-eosin staining showed that the cross-sectional area and diameter of muscle fibers of each skeletal muscle were smaller in the other four groups than that in the sham group (P < 0.05). The cross-sectional area and diameter of muscle fibers of each skeletal muscle in the NT3-chitosan combined rehabilitation training group were higher than the lesion control group, the lesion control combined rehabilitation training group, and NT3-chitosan group (P < 0.05). (3) At 12 weeks after operation, the acetylcholinesterase staining showed that the average optical density of the acetylcholinesterase on motor endplate of the muscle was lower in the other four groups than that in the sham group (P < 0.05); the average optical density of the acetylcholinesterase of the motor endplate in the NT3-chitosan combined rehabilitation training was significantly higher than that in the lesion control, lesion control combined rehabilitation training, and NT3-chitosan groups (P < 0.05). (4) The results show that NT3-chitosan combined with rehabilitation training can effectively prevent muscular atrophy of hind limb skeletal muscles in rats with complete spinal cord injury, improve the average optical density of the acetylcholinesterase of the motor endplate, reduce neuromuscular joint degeneration, and improve rat hindlimb motor function.

Objective To investigate the effect of C peptide on tubulointerstitial oxidative stress injury in rats with diabetic nephropathy (DN) induced by streptozotocin (STZ). Methods Wistar rats with DN induced by STZ were randomly divided into control group and C peptide group. All rats were treated via intraperitoneal micro-osmotic pump. Normal Wistar rats at the same age were used as the normal group. Blood glucose and24 h urinary albumin were measured before treatment and every 4 weeks during treatment period. After 12 weeks of treatment, the expression of transforming growth factor - β1 (TGF - β1) in renal tubulointerstitium was detected by immunohistochemical staining. Fresh tubulointerstitial tissue homogenate was harvested and the activity of superoxide dismutase (SOD) was detected in WST-1 method, the content of malondialdehyde (MDA) was detected in TBA method, and the expression of protein kinase A (PKA) mRNA was detected by Real-time PCR. Results Levels of blood glucose, 24 h urinary albumin, MDA content and the expression of TGF-β1 were higher in the control group than those in the normal group, while SOD activity decreased and PKA mRNA was downregulated. C peptide treatment did not change blood glucose level but slowed down the increasing of 24 h urinary albumin, reduced the tubulointerstitial TGF-β1 expression and MDA content, increased the SOD activity and upregulated the PKA mRNA expression. Conclusions C-peptide can decrease the tubulointerstitial oxidative stress in DN rats by activating PKA pathway and then improve tubulointerstitial fibrosis and attenuate proteinuria.

OBJECTIVE:To observe therapeutic efficacy and safety of Danshen chuanxiongqin injection combined with flunari-zine hydrochloride in the benign prevention and treatment of paroxysmal positional vertigo (BPPV) and lower extremity deep ve-nous thrombosis (DVT) in post-operative long-term bedridden patients with lower limb fractures. METHODS:300 post-operative long-term bedridden patients with lower limb fractures were selected and randomly divided into observation group and control group,with 150 cases in each group. Control group was given Flunarizine hydrochloride capsules orally 10 mg,qd;observation group was additionally given Danshen chuanxiongqin injection 10 ml+5% Glucose injection 250 ml,ivgtt,qd. The incidence of BPPV and DVT were observed in 2 groups after intervention,and the circumference of lower limb,blood coagulation indexes, blood rheology indexes and inflammatory factor were observed before and after intervention,and the incidence of ADR was com-pared. RESULTS:The incidence of BPPV and DVT in observation group were 18.0% and 16.7%,which were significantly lower than in control group(48.7% and 52.7%),with statistical significance(P<0.05);after intervention,the circumference of lower limb,blood rheology indexes and the levels of inflammatory factors in 2 groups were decreased significantly, while the coagula-tion indicators were significantly improved;the observation group was better than the control group,with statistical significance (P<0.05). There was no statistical significance in the incidence of ADR between 2 groups(P>0.05). CONCLUSIONS:Danshen chuanxiongqin injection combined with flunarizine hydrochloride is effective in the prevention of BPPV and DVT in long-term bed-ridden patients with lower limb fractures,with low incidence of ADR.

Objective To introduce a community-based rehabilitation(CBR) network model in Shijiazhuang,Hebei province.Methods The work model in Qiaodong district in Shijiazhuang city was evaluated according to the national CBR standard and analyzed.Results Their work has met the national standard,the score of management section was the highest among all areas.Conclusion The work was featured by government dominate,strong management network and technical support.It implied the social model of CBR.

Objective:To construct the post competency index system of the standardized training of traditional Chinese medicine(TCM)residents, and to provide reference for the curriculum design of TCM residents′ standardized training.Methods:Based on the analysis of relevant literature, 22 experts engaged in clinical, teaching or administrative management of TCM residential training were consulted by Delphi method from August to December 2019, and the evaluation indexes were screened and the importance was assigned. Then, analytic hierarchy process was used to determine the weight of each index. In January 2020, the index system was tested by questionnaire survey.Results:The competency evaluation index system of TCM residents was composed of 7 first-level indexes and 61 second-level indexes. The first-level indexes included occupational values and professionalism(weight: 0.208 2), clinical skills(weight: 0.208 2), mastery and application of medical knowledge and related knowledge(weight: 0.208 2), communication skills(weight: 0.198 4), management and teamwork ability(weight: 0.114 1), critical thinking and learning research ability(weight: 0.033 4), group health and health systems(weight: 0.029 5). The total Cronbach α coefficient of the self-evaluation questionnaire designed according to the evaluation index system was 0.976. The results of KMO test and Bartlett spherical test respectively were 0.954 and 0.000, indicating that the questionnaire had high reliability and validity, and the evaluation index system could reflect the post competency level of TCM residents. Conclusions:The index system of TCM resident competency is highly reliable. TCM resident training should strengthen the cultivation of TCM clinical thinking, and urgently need to supplement the content of medical humanities education.

T cells engineered with chimeric antigen receptor (CAR) have been successfully applied to treat advanced refractory B cell malignancy.However,many challenges remain in extending its application toward the treatment of solid tumors.The immunosuppressive nature of tumor microenvironment is considered one of the key factors limiting CAR-T efficacy.One negative regulator of T cell activity is lymphocyte activation gene-3 (LAG-3).We successfully generated LAG-3 knockout T and CAR-T cells with high efficiency using CRISPR-Cas9 mediated gene editing and found that the viability and immune phenotype were not dramatically changed during in vitro culture.LAG-3 knockout CAR-T cells displayed robust antigen-specific antitumor activity in cell culture and in murine xenograft model,which is comparable to standard CAR-T cells.Our study demonstrates an efficient approach to silence immune checkpoint in CAR-T cells via gene editing.

Objective:To explore relationship between serum 25-hydroxy vitamin D ( 25OHD ) level and ketosis-prone in patients with newly-diagnosed type 2 diabetes mellitus(T2DM).Methods:One hundred and thirty-four patients with newly diagnosed T2DM (103 males, 31 females) admitted in The Third Affiliated Hospital, Southern Medical University from January 2017 to January 2019 were enrolled into this study, including 36 patients with ketosis-prone (KPDM group) and 98 patients without ketosis(NKPDM group). Clinical characteristics, including height, weight, and history of hypertensive disease were collected. Serum 25OHD levels, lipid profile, islet function and glycosylated hemoglobin (HbA 1C)levels, ICA, IAA, GAD-Ab, etc., were measured. Results:Among the 134 patients, the patients with vitamin D deficiency, insufficiency, and sufficiency were 71 cases(52.99%), 52 cases(38.81%), and 11 cases(8.20%), respectively. KPDM group had significantly lower serum 25OHD level than NKPDM group[(44.12±9.77) nmol/L vs (55.35±15.31) nmol/L, P<0.01]. The rate of vitamin D deficiency was significantly higher in KPDM group compared to that in NKPDM group [(77.78% vs 43.88%), P<0.01]. The prevalence of KPDM varied significantly in different vitamin D status groups( P<0.01). Logistic regression analysis suggested that low serum 25OHD, younger age, high HbA 1C, and triglyceride were risk factors to ketosis-prone in newly diagnosed T2DM( P<0.01). Conclusion:Vitamin D deficiency is a common problem in newly diagnosed T2DM, especially in KPDM. Low serum 25OHD level seems to be an independent risk factor for ketosis-prone in patients with newly diagnosed T2DM.

Objective:To analyze the association between the pre-pregnancy oral microbiota of women and fetal overgrowth, and the possible mechanisms involved.Methods:A nested case-control study design based on a pre-pregnancy cohort was used to select 51 mothers who delivered macrosomia and/or large-for-gestational-age (LGA) infants from the population recruited at the Maternal and Child Health Care Hospital of Jiading District in Shanghai from October 2016 to December 2021 as the case group. A control group was formed by selecting 204 mothers who delivered infants with normal birth weight and appropriate for gestational age during the same period, in a 1:4 ratio. The LGA subgroup consisted of 48 mothers who delivered LGA infants from the total population, and a corresponding control group of 192 was randomly selected from the remaining mothers who delivered non-LGA infants in a 1∶4 ratio for the LGA subgroup analysis. The 16S rRNA gene sequencing technique was utilized to detect pre-pregnancy saliva samples to compare the characteristics of the oral microbiota, differential microorganisms, and differential functional pathways between groups. Nonparametric Wilcoxon rank-sum tests, two independent samples t-tests, or Chi-square (or Fisher's exact) tests were used for statistical analysis. Factor analysis was conducted on the pre-pregnancy diet data of women, and the primary dietary pattern of each study subject was identified based on the highest score of the dietary pattern factors. For microbiota count data, α and β diversity indices were calculated using R and QIIME2 software, and the corresponding microbiota functional count data were acquired through PICRUSt2. Results:(1) General data: There was no significant difference in the time interval from pre-pregnancy sampling to pregnancy and from sampling to delivery between the two groups. In the case group, there were three cases of macrosomia and 48 cases (94.1%) of LGA. The corresponding control group for the LGA subgroup consisted of 192 cases. There were no significant differences in dietary patterns between the case group and the control group. (2) α diversity analysis: The species richness index of the case group was lower than that of the control group [(367.27±84.57) vs. (408.71±93.08), multivariate analysis, P=0.009], while no significant differences were found between the two groups in the Shannon and Simpson indices; the species richness index of the LGA subgroup was also lower than that of the corresponding control group [(371.04±83.92) vs. (408.04±94.21), multivariate analysis, P=0.033], with no significant differences in the Shannon and Simpson indices. (3) β diversity analysis: There was a statistically significant difference in the unweighted UniFrac distance of the oral microbiota between the case group and the control group ( R2=0.006, F=1.479, P=0.048). No significant differences were found in the β diversity indices of the oral microbiota between the LGA subgroup and the corresponding control group. (4) Differential microbiota analysis: There were 14 differential microbiotas from phylum to genus between the case group and the control group. At the genus level, members of the G1 genus of the Streptococcaceae were enriched in the case group, while the Lautropia, Dialister, Leptotrichia, and Rothia were enriched in the control group. In the LGA subgroup and its corresponding control group, there were 14 differential microbiota from phylum to genus; at the genus level, Leptotrichia, Rothia, G6 genus of the Saccharibacteria, and Selenomonas were enriched in the control group (all LDA value>2, and all P<0.05). (5) Differential functional analysis: In the case group, metabolic pathways such as nicotinate degradation [log 2 fold change ( FC)=3.510, q=0.005], de novo synthesis of pyrimidine nucleotides (log 2FC=0.078, q=0.005), and L-tyrosine degradation pathway (log 2FC=0.710, q=0.034) were enriched in the oral microbiota of women. In the LGA subgroup, compared to the corresponding control group, metabolic pathways related to nicotinate degradation were enriched in the oral microbiota (log 2FC=3.660, q=0.012). Conclusions:There are differences in the structure of the pre-pregnancy oral microbiota of mothers with overgrown fetuses compared to those with normally grown fetuses, and mothers of normally grown fetuses show higher diversity in their pre-pregnancy oral microbiota. The enrichment of certain pathogenic bacteria and the reduction of symbiotic bacteria in the pre-pregnancy oral microbiota are associated with fetal overgrowth, and this association may be mediated by functional pathways such as nicotinate degradation.

As a member of the AFF (AF4/FMR2) family, AFF4 is a transcription elongation factor that is a component of the super elongation complex. AFF4 serves as a scaffolding protein that connects transcription factors and promotes gene transcription through elongation and chromatin remodelling. Here, we investigated the effect of AFF4 on human dental follicle cells (DFCs) in osteogenic differentiation. In this study, we found that small interfering RNA-mediated depletion of AFF4 resulted in decreased alkaline phosphatase (ALP) activity and impaired mineralization. In addition, the expression of osteogenic-related genes (DLX5, SP7, RUNX2 and BGLAP) was significantly downregulated. In contrast, lentivirus-mediated overexpression of AFF4 significantly enhanced the osteogenic potential of human DFCs. Mechanistically, we found that both the mRNA and protein levels of ALKBH1, a critical regulator of epigenetics, changed in accordance with AFF4 expression levels. Overexpression of ALKBH1 in AFF4-depleted DFCs partially rescued the impairment of osteogenic differentiation. Our data indicated that AFF4 promoted the osteogenic differentiation of DFCs by upregulating the transcription of ALKBH1.
Biomarkers ; metabolism ; Cell Differentiation ; Cells, Cultured ; Dental Sac ; drug effects ; metabolism ; Gene Expression Regulation ; Humans ; Osteogenesis ; genetics ; Repressor Proteins ; Transcription Factors ; genetics ; metabolism ; Transcriptional Elongation Factors ; metabolism