Objective: To study the anti-tumor efficacy induced by antibodized tumor epitope PDTRP gene immunization. Methods: Three copies of tumor associate gene PDTRP from MUCI tandem repeats were designed and mimicked the conformation of MUCI by Insight Ⅱ . The ?lneo-PDTRP plasmid was further constructed, in which the PDTRP target gene was inserted into CDR3 of the ?1 -neo vector. The specific humoral and cellular immune responses towards to PDTRP were detected after intraspleen immunized Balb/c mice with "ylneo-PDTRP. And the immune protection assay was also done to observe whether the mice immunized with ?lneo-PDTRP could prolong the survival after tumor challenge. Results: The conformation of three copies of PDTRP mimicked the conformation of MUCI tandem repeats. The expression of ?lneo-PDTRP could be detected after in vitro transfect. The specific antibody against PDTRP epitope could be induced and increase to a higher titer after intraspleen injection with a ?lneo-PDTRP plasmid. And the specific proliferation and cytotox-ic function of lymphocyte were also increased. There is a significant survival from mice immunized with ?lneo-PDTRP a-gainst the 4T1-PDTRP tumor challenge. Conclusions: Gene immunization with ?lneo-PDTRP could elicit both humoral and cellular tumor specific immune response and had the protective effect.

Objective To investigate the effects of glycosaminoglycans (HGAG) on the immune function of peripheral blood cells from patients with pulmonary tuberculosis.Methods Peripheral blood monouclear cells (PBMC) were isolated from peripheral blood of 40 healthy people (healthy group) and 30 tuberculosis patients (tuberculosis group) and cocultured with HGAG in vitro for 24 hours.Flow cytometry was used to detect the expression of CD45RA and CD45RO,as well as the expression of CD1a and CD83.Results The results showed that the expression of CD45RA and CD45RO in the tuberculosis group was the most significant (P ＜ 0.05) at the concentration of 50 μg/m coculturing with HGAG.The expression of CD45RA and CD45RO were most obvious in the healthy group at the concentration of 10 μg/ml and 50 μg/ml respectively (P ＜0.001).The difference of CD45RA between the two groups was no significant (P ＞0.05),while the difference of CD45RO was statistically significant (P ＜ 0.01) before co-culturing.The expression of CD45RA and CD45RO at 10 μg/ml and 50 μg/ml after co-culturing with HGAG were statistically significant (P ＜ 0.05).There was no statistical difference in CD1a and CD83 in healthy group before and after co-culturing (P ＞ 0.05),while there was statistically difference (P ＜ 0.05) before and after culturing in tuberculosis group.Before co-culturing,there was no significant difference in the expression of CD1a between the healthy group and the tuberculosis group (P ＞ 0.05),but CD83 expression was statistically different (P ＜ 0.001).After co-culturing,there were no significant differences in CD1a and CD83 expression between healthy and healthy groups (P ＞ 0.05).Conclusions HGAG can down-regulate the expression of CD45RA and up-regulate the expression of CD45RO in a certain concentration range,and promote the maturation of dendritic cells (DC) in tuberculosis patients and regulate the cellular immunity of patients with pulmonary tuberculosis in vitro.

Objective To investigate the effects of sodium butyrate on the activity of RAW264.7 cells and the osteoclast differentiation.Methods The RAW264.7 cells were treated by sodium butyrate at concentrations of 0,0.25,0.50,1.00,2.00,3.00,4.00 and 5.00 mmol/L,with 3 double pores for each concentration.The cytotoxicity of sodium butyrate on RAW264.7 cells was detected by a CCK-8 kit.The effects of sodium butyrate (0,0.25,0.50 and 1.00 mmol/L) on apoptosis of RAW264.7 cells were detected by Hoechst33342 staining.RAW264.7 cells were induced into osteoclasts by osteoclast differentiation factors.The experiment was carried out in 2 groups (n =3).After induced maturation,the experimental group was treated with 1.00 mmol/L sodium butyrate and the control medium was added only with the same volume of solvent.The number of osteoclasts and the area of bone resorption were observed and compared.The differentiation of RAW264.7 cells was detected by tartrate-resistant acid phosphatase (TRAP) staining.Western blotting was used to detect the effects of sodium butyrate (0,0.25,0.50 and 1.00 mmol/L) on NF-κB-related signaling pathway in RAW264.7 cells.Results Compared with the group of 0 mmol/L sodium butyrate,the activity of cells treated with 1.00,2.00,3.00,4.00 and 5.00 mmol/L sodium butyrate for 24 h was significantly decreased (P ＜ 0.05).Treatment with 1.00 mmol/L sodium butyrate for 24 h induced apoptosis.The number of osteoclasts in the control group and the experimental group were 9.33 ± 2.08 and 4.67 ± 1.16,respectively,showing a significant difference between the 2 groups (t =3.395,P =0.027).The percentages of bone resorption area in the control group and the experimental group were 52.43％ ± 5.38％ and 14.28％ ± 2.72％,respectively,also showing a significant difference between the 2 groups (t =10.970,P ＜ 0.001).Western blot results showed that,compared with other concentrations of sodium butyrate,treatment with 1 mmol/L sodium butyrate on RAW264.7 cells for 24 h led to an increase in the expression levels of cytoplasmic p65,B lymphoma-2 associated X protein and cleaved-caspase 3 and the acetylation of Histone H3 but a decrease in the phosphorylation level of α/β subunit of NF-κB kinase.Conclusions With the increased concentration of sodium butyratecan,the activity of NF-κB may be suppressed and the number of apoptotic cells may increase.1.00 mmol/L sodium butyrate can reduce osteoclast formation and bone resorption area.

Objective　To investigate the prevalence,age distribution and risk factors of cerebral small vessel disease (CSVD) neuroimaging markers in Shanghai elderly community population.Methods　From August 2016 to October 2019,subjects aged above 50 years and qualified for the study were enrolled from Jingan,Qingpu and Minghang districts in Shanghai.Uniformly designed clinical research forms,neuropsychological assessment,laboratory tests and MRI were performed to gather information and to analyze the prevalence,severity and distribution of lacune,white matter hyperintensity (WMH),cerebral microbleed (CMB) and perivascular space (PVS).Associations between risk factors and neuroimaging markers were analyzed by multivariate logistic regression models.Results　 Among 870 enrolled subjects who had undergone MRI,median age was 68 years and 40.2% male.Lacune,WMH (Fazekas≥2),CMB and PVS were detected in 17.6%,29.4%,27.8%,96.5% of subjects respectively.With every 10 years’increase in age,there was a 35% increase in lacune,126% increase in WMH and 51% increase in CMB.Compared with non-hypertensive subjects,hypertensive subjects had a 1.98-fold of risk detecting lacune on MRI,while a 1.64,1.91,1.45 fold of risk for WMH,CMB and PVS.ConclusionsAge and hypertension are the major risk factors for CSVD in elderly community population.Age-stratified prevalence of single imaging marker and total CSVD score may have a clinical significance in diagnosing CSVD.

Background and Objectives: Epithelial-Mesenchymal transition (EMT) is one of the origins of myofibroblasts in renal interstitial fibrosis. Mesenchymal stem cells (MSCs) alleviating EMT has been proved, but the concrete mechanism is unclear. To explore the mechanism, serum-free MSCs conditioned medium (SF-MSCs-CM) was used to treat rat renal tubular epithelial cells (NRK-52E) fibrosis induced by transforming growth factor-β1 (TGF-β1) which ameliorated EMT. Methods: and Results: Galectin-3 knockdown (Gal-3 KD) and overexpression (Gal-3 OE) lentiviral vectors were established and transfected into NRK-52E. NRK-52E fibrosis model was induced by TGF-β1 and treated with the SF-MSCs-CM for 24 h after modelling. Fibrosis and autophagy related indexes were detected by western blot and immunocytochemistry. In model group, the expressions of α-smooth muscle actin (α-SMA), fibronectin (FN), Galectin-3, Snail, Kim-1, and the ratios of P-Akt/Akt, P-GSK3β/GSK3β, P-PI3K/PI3K, P-mTOR/mTOR, TIMP1/MMP9, and LC3B-II/I were obviously increased, and E-Cadherin (E-cad) and P62 decreased significantly compared with control group. SF-MSCs-CM showed an opposite trend after treatment compared with model group. Whether in Gal-3 KD or Gal-3 OE NRK-52E cells, SF-MSCs-CM also showed similar trends. However, the effects of anti-fibrosis and enhanced autophagy in Gal-3 KD cells were more obvious than those in Gal-3 OE cells. Conclusions SF-MSCs-CM probably alleviated the EMT via inhibiting Galectin-3/Akt/GSK3β/Snail pathway. Meanwhile, Gal-3 KD possibly enhanced autophagy via inhibiting Galectin-3/Akt/mTOR pathway, which synergistically ameliorated renal fibrosis. Targeting galectin-3 may be a potential target for the treatment of renal fibrosis.

【Objective】 To explore the effect of metaphors based on children′s games on the negative emotion and behavior of children during the recovery period of traumatic brain injury, in order to provide reference for making intervention strategy for these children. 【Methods】 From May 2021 to October 2022, a total of 60 children aged 6-12 years who underwent rehabilitation in the rehabilitation center of the Second Affiliated Hospital of Wenzhou Medical University were selected into this study, and were randomly divided into experimental group and control group, with 30 cases in each group.The control group was given routine nursing, while the experimental group was given metaphorical intervention based on children′s games.The scores of social living ability, abnormal behavior detection rate, depression score, anxiety score and cognitive function were compared between the two groups. 【Results】 The scale score of the two groups was not significantly different before intervention(P>0.05).After intervention, the infant-junior high school students′ social living ability score, Mini-Mental State Examination and the Loewenstein Occupational Therapy Cognitive Assessment score in the experimental group were higher than those in the control group (t=3.175, 5.265, 7.835, P<0.001).The scores of Child Behavior Checklist (CBCL), the children′s depression inventory and the screen for child anxiety related emotional disorders in the experimental group were lower than those in the control group (Z=7.823, t=6.398, 6.248, P<0.001). 【Conclusions】 Metaphorical intervention based on children′s games can reduce the negative emotions of anxiety and depression in children with traumatic brain injury, improve their social life ability, reduce the CBCL score, and improve the cognitive function.

ObjectiveTo explore the possible mechanism of Bimin Formula （鼻敏方） in treating lung-spleen qi deficiency syndrome of allergic rhinitis （AR） with high mucin secretion. MethodsThirty-four SD rats were randomly divided into a blank group （8 rats）， a model group （8 rats）， a low-dose Bimin Formula group （8 rats）， and a high-dose Bimin Formula group （10 rats）. Except for the blank group， the other groups were subjected to AR lung-spleen qi deficiency rat models induced by smoking， gavage of Ginkgo biloba leaf extract， and ovalbumin. After modeling， rats in the low- and high-dose Bimin Formula groups were given Bimin Formula concentrate （concentration of 2.16 g/ml） by gavage at doses of 1.08 g/100 g and 2.16 g/100 g， respectively， while rats in the model group were given 0.5 ml/100 g of normal saline by gavage， once daily for 28 days； the blank group was not intervened. Behavioral assessments were performed after intervention. ELISA was used to detect the levels of peripheral blood total immunoglobulin E （IgE）. HE staining was used to observe the pathological changes of nasal mucosa epithelium in rats， while immunohistochemistry was used to detect the expression of transmembrane protein 16A （TMEM16A） and mucin 5AC （MUC5AC） protein in nasal mucosa. Western Blot was used to detect the expression of nuclear factor kappa B （NF-κB） protein， and RT-PCR was used to detect the expression of TMEM16A， MUC5AC， and NF-κB mRNA in nasal mucosa. ResultsHE staining showed that the nasal mucosa epithelial cell structure in the blank group was intact without shedding， swelling， or necrosis； the nasal mucosa epithelial tissue of rats in the model group was thickened and partially shed， with infiltration of eosinophils and lymphocytes visible； the pathological changes in nasal mucosa tissue of rats in the high- and low-dose Bimin Formulagroups were improved， and more improvement was showen in the high-dose group. Compared with those in the blank group， the behavioral scores and peripheral blood total IgE levels of rats in the model group significantly increased， as well as the expression of TMEM16A， MUC5AC， and NF-κB proteins and mRNA in nasal mucosa （P＜0.05 or P＜0.01）. Compared with those in the model group， the behavioral scores and peripheral blood total IgE levels of rats in the high-dose Bimin Formula group decreased， and the expression of TMEM16A， MUC5AC， and NF-κB proteins and mRNA in nasal mucosaalso decreased （P＜0.05 or P＜0.01）； the behavioral scores and peripheral blood total IgE levels of rats in the low-dose Bimin Formula group were reduced， and the expression of TMEM16A and MUC5AC proteins and mRNA in nasal mucosa， as well as the expression of NF-κB protein decreased （P＜0.05 or P＜0.01）， but the difference in NF-κB mRNA expression was not statistically significant （P＞0.05）. Compared with the low-dose Bimin Formula group， the expression of NF-κB protein in the high-dose group decreased （P＜0.01）. ConclusionBimin Formula may improve the symptoms and high mucus secretion of AR lung-spleen qi deficiency by regulating the TMEM16A/NF-κB/MUC5AC signaling pathway in nasalmucosa.

Objective: To evaluate the effect of 3D-printed oval root canal preparation by using small field-of-view cone beam CT (CBCT) combined with computer-aided technologies. Methods: An extracted tooth with a suitable single oval canal was selected by small field-of-view CBCT scanning. Three-dimensional (3D)-printed resin teeth were obtained based on the CBCT data after data conversion and processing. 50 general dentists were selected to prepare the oval root canal of the resin teeth with X-Gold rotary Ni-Ti instruments. Small field-of-view CBCT was applied to scan the oval root canals before and after preparation. Then, computer-aided technologies were used to calculate and compare these CBCT data, analyzing the effects of oval root canal cleaning as well as the root canal deviation by 3D reconstruction and rendering of the images. Results : Among the 50 cases, the mean unprepared area of the oval root canal wall was（56.20 ± 11.91）% and the mean maximum deviation distance of the root canal was（0.140 ± 0.041）mm. There was no correlation between root canal cleanliness and deviation (r=0.18, P=0.212). Conclusion Combined with small field-of-view CBCT and computer-aided technology, we can effectively quantitatively evaluate the root canal preparation effect.