BACKGROUND:As tissue engineering brings new hope to the worldwide problem of articular cartilage repair,the construction of light-curing 3D printed hydrogel scaffolds with biomimetic composition is of great significance for cartilage tissue engineering. OBJECTIVE:To construct a biomimetic methacryloylated hyaluronic acid/acellular Wharton's jelly composite hydrogel scaffold by digital light processing 3D printing technology,and to evaluate its biocompatibility. METHODS:Wharton's jelly was isolated and extracted from human umbilical cord,then decellulated,freeze-dried,ground into powder,and dissolved in PBS to prepare 50 g/L acellular Wharton's jelly solution.Methylallylated hyaluronic acid was prepared,lyophilized and dissolved in PBS to prepare 50 g/L methylallylated hyaluronic acid solution.Acellular Wharton's jelly solution was mixed with methacrylyacylated hyaluronic acid solution at a volume ratio of 1:1,and was used as bio-ink after adding photoinitiator.Methylacrylylated hyaluronic acid hydrogel scaffolds(labeled as HAMA hydrogel scaffolds)and methylacrylylated hyaluronic acid/acellular Wharton's jelly gel scaffolds(labeled as HAMA/WJ hydrogel scaffolds)were prepared by digital light processing 3D printing technology,and the microstructure,swelling performance,biocompatibility,and cartilage differentiation performance of the scaffolds were characterized. RESULTS AND CONCLUSION:(1)Under scanning electron microscope,the two groups of scaffolds showed a three-dimensional network structure,and the fiber connection of HAMA/WJ hydrogel scaffold was more uniform.Both groups achieved swelling equilibrium within 10 hours,and the equilibrium swelling ratio of HAMA/WJ hydrogel scaffold was lower than that of HAMA hydrogel scaffold(P＜0.05).(2)CCK-8 assay showed that HAMA/WJ hydrogel scaffold could promote the proliferation of bone marrow mesenchymal stem cells compared with HAMA hydrogel scaffold.Dead/live staining showed that bone marrow mesenchymal stem cells grew well on the two groups of scaffolds,and the cells on the HAMA/WJ hydrogel scaffolds were evenly distributed and more cells were found.Phalloidine staining showed better adhesion and spread of bone marrow mesenchymal stem cells in HAMA/WJ hydrogel scaffold than in HAMA.(3)Bone marrow mesenchymal stem cells were inoculated into the two groups for chondrogenic induction culture.The results of qRT-PCR showed that the mRNA expressions of agglutinoglycan,SOX9 and type Ⅱ collagen in the HAMA/WJ hydrogel scaffold group were higher than those in the HAMA hydrogel scaffold group(P＜0.05,P＜0.01).(4)These findings indicate that the digital light processing 3D bioprinting HAMA/WJ hydrogel scaffold can promote the proliferation,adhesion,and chondrogenic differentiation of bone marrow mesenchymal stem cells.

OBJECTIVE To confirm the therapeutic effect of Jintiange capsules on osteoarthritis(OA) and the potential mechanism of synovial mesenchymal stem cell exosomes(SMSC-Exos) and articular chondrocytes(ACs) in the treatment of OA based on high-throughput sequencing technology. METHODS Type Ⅱ collagenase-induced OA rats were used for efficacy verification through general behavioral observation, bipedal balance difference experiment, mechanical foot reflex threshold, Micro-CT observation, and Safranin O-Fast Green staining. SMSCs and ACs were cultured in suitable concentration of drug-containing serum, and mRNA sequencing was performed on ACs in the control, model, and Jintiange capsules groups, as well as miRNA sequencing on SMSC-Exos. Differential expressed mRNAs and miRNAs were screened and target genes were predicted. The common differential expressed genes between SMSC and ACs were obtained by intersecting the differential expressed genes, and a miRNA-mRNA regulatory network was constructed using Cytoscape software. The expression trend analysis of common differential expressed genes was conducted, as well as the correlation analysis between differential expressed gene mRNA and miRNA, Micro-CT efficacy indicators, and differential expressed gene mRNA. RESULTS Under the pathological state of OA, the expression of miRNA-23a-3p, miRNA-342-3p, miRNA-146b-5p, miRNA-501-3p, and miRNA-214-3p were down-regulated, while miRNA-222-3p, miRNA-30e-3p, miRNA-676-3p, and miRNA-192-5p were up-regulated (P<0.05). The expressions of these miRNAs were significantly reversed after intervention with drug-containing serum of Jintiange capsules. There was a certain correlation between Micro-CT efficacy indicators, mRNA and miRNA. CONCLUSION Jintiange capsule has obvious efficacy in the treatment of OA, and its mechanism may be related to the promotion of SMSC-Exos targeting ACs to transport miRNA and then regulate Serpinb10, Ntn1, Il1b, Tgm2, Megf10, Il11, Cd40, Slc15a3, Pou2f2 and other genes.

Objective:To implement evidence-based practice of discharge preparation service in maternal and to explore the clinical effect.Methods:Based on the evidence-based continuous quality improvement framework, apply the evidence on the clinical through evidence acquisition, baseline evaluation, evidence introduction, and after-effect evaluation. Sixty women hospitalized in the obstetrics department of the Affiliated Hospital of Chengde Medical College from January to March 2021 were selected as the baseline group, and 60 women hospitalized in the obstetrics department of the hospital from April to July 2021 were selected as the evidence application group, to evaluate and compare the quality of discharge guidance for practitioners, discharge readiness level, maternal and newborn management knowledge level, incidence of maternal and newborn complications and the compliance of practitioners to evidence.Results:Finally, 10 pieces of evidence applied to clinical practice were included and 15 review indicators were translated. After the application of evidence, the quality of discharge guidance of practitioners increases from (141.83±24.66) to (166.13±10.30), the maternal discharge readiness increased from (127.12 ± 26.29) to (137.63 ± 22.59) , the score of maternal knowledge level increased from (164.58 ± 20.10) to (176.08 ± 24.25) , t values were -7.04, -2.39 and -2.83, all P<0.05. The incidence of postpartum women breast disease, postnatal constipation and newborn eczema, newborn red buttocks, newborn diarrhea, newborn jaundice decreased significantly, the difference were statistically significant, χ2 values were 5.21-7.50, all P<0.05. After the application of evidence, the implementation rate of the review indicators increased from ≤31.67% to ≥86.66%. Conclusions:The evidence-based practice of discharge preparation service in postpartum women improve the quality of discharge guidance for practitioners and compliance with evidence, promote clinical quality improvement, can be used for clinical reference.

Objective:To evaluate the regulatory role and potential mechanism of Urease B(UreB) on macrophages.Methods:Bone marrow-derived macrophages (M0) were stimulated by recombinant UreB protein and then flowcytometry and ELISA were used to detect the apoptosis, polarization and antigen presentation-related biomarkers expression. CD4 + T cell co-culture assay, CFSE stain and flowcytometry were used to evaluate the impacts of UreB on antigen presentation capacity of macrophages. Truncated UreB protein, NanoBiT assay and co-immunoprecipitation were used to identify the binding sites of UreB to TLR2. Results:UreB promoted apoptosis and skewed macrophages from M1 to M2 in the presence of M1-inducer LPS. Moreover, UreB inhibited the expression of antigen presentation biomarkers, MHCⅡ and CD86 on macrophages, and further inhibited the proliferation and IFN-γ expression of CD4 + T cells. Molecular analyses revealed that the binding between seven carboxy-terminal amino acid residues of UreB and TLR2 were required for the UreB-mediated inhibitory effects. Conclusions:The findings in this study demonstrate that UreB mainly depends on the binding between seven carboxy-terminal amino acid residues and TLR2 to perform immune-suppressive activities, and which may provide valuable information for the design and optimization of UreB-based vaccines against Helicobacter pylori infection.

Objective:To examine the correlation factors of contralateral central lymph node metastasis in unilateral papillary thyroid carcinoma with lateral cervical lymph node metastasis.Methods:The clinical data of patients with unilateral papillary thyroid carcinoma who underwent total thyroidectomy and bilateral central lymph node dissection and ipsilateral cervical lymph node dissection from June 2016 to June 2018 at Department of Thyroid Surgery, West China Hospital, Sichuan University were analyzed retrospectively. A total of 317 patients, including 87 males and 230 females, aged (41.4±12.1) years (range: 16 to 75 years), were enrolled in this study. The risk factors of contralateral central lymph node metastasis were analyzed by χ 2 test and Spearman correlation analysis. Results:There were 116, 69, 269, and 181 cases of pretracheal lymph node, prelaryngeal lymph node, ipsilateral central lymph node and contralateral central lymph node metastasis, respectively, and 16 cases of skipping metastasis. Univariate analysis showed that contralateral central lymph node metastasis was associated with gender, maximum tumor diameter, capsule invasion, pretracheal lymph node metastasis, prelaryngeal lymph node metastasis, and ipsilateral central lymph node metastasis (all P<0.05). Spearman correlation analysis showed that male ( r s=0.162, P=0.004), maximum tumor diameter>10 mm ( r s=0.184, P=0.001), capsule invasion ( r s=0.135, P=0.016), pretracheal lymph node metastasis ( r s=0.394, P<0.01), prelaryngeal lymph node metastasis ( r s=0.272, P<0.01) and ipsilateral central lymph node metastasis ( r s=0.203, P<0.01) were independent correlation factors for contralateral central lymph node metastasis. Conclusion:For patients with unilateral papillary thyroid carcinoma with ipsilateral cervical lymph node metastasis, bilateral central lymph node dissection should be considered if male, tumor diameter>10 mm, capsule invasion, pretracheal lymph node metastasis, prelaryngeal lymph node metastasis, or ipsilateral central lymph node metastasis.

Objective:To examine the correlation factors of contralateral central lymph node metastasis in unilateral papillary thyroid carcinoma with lateral cervical lymph node metastasis.Methods:The clinical data of patients with unilateral papillary thyroid carcinoma who underwent total thyroidectomy and bilateral central lymph node dissection and ipsilateral cervical lymph node dissection from June 2016 to June 2018 at Department of Thyroid Surgery, West China Hospital, Sichuan University were analyzed retrospectively. A total of 317 patients, including 87 males and 230 females, aged (41.4±12.1) years (range: 16 to 75 years), were enrolled in this study. The risk factors of contralateral central lymph node metastasis were analyzed by χ 2 test and Spearman correlation analysis. Results:There were 116, 69, 269, and 181 cases of pretracheal lymph node, prelaryngeal lymph node, ipsilateral central lymph node and contralateral central lymph node metastasis, respectively, and 16 cases of skipping metastasis. Univariate analysis showed that contralateral central lymph node metastasis was associated with gender, maximum tumor diameter, capsule invasion, pretracheal lymph node metastasis, prelaryngeal lymph node metastasis, and ipsilateral central lymph node metastasis (all P<0.05). Spearman correlation analysis showed that male ( r s=0.162, P=0.004), maximum tumor diameter>10 mm ( r s=0.184, P=0.001), capsule invasion ( r s=0.135, P=0.016), pretracheal lymph node metastasis ( r s=0.394, P<0.01), prelaryngeal lymph node metastasis ( r s=0.272, P<0.01) and ipsilateral central lymph node metastasis ( r s=0.203, P<0.01) were independent correlation factors for contralateral central lymph node metastasis. Conclusion:For patients with unilateral papillary thyroid carcinoma with ipsilateral cervical lymph node metastasis, bilateral central lymph node dissection should be considered if male, tumor diameter>10 mm, capsule invasion, pretracheal lymph node metastasis, prelaryngeal lymph node metastasis, or ipsilateral central lymph node metastasis.

To assess the performance of radiomics model based on cardiac magnetic resonance imaging (CMR) cine sequence for assessing the severity of mitral regurgitation.
 Methods: A total of 80 patients who underwent CMR and echocardiography examination were retrospectively enrolled, including 67 patients with no or slight mitral regurgitation and 13 patients with moderate or severe mitral regurgitation. The relative difference in average filtered gradient (RDAFG) of CMR cine sequence were generated, which were combined with minimum output sum of squared error tracker (MOSSE) to extract 25 radiomics features. After reducing feature dimensionality by principal component analysis (PCA) and oversampling the minority samples, the radiomics model was established using support vector machine (SVM). The performance of the model was assessed by receiver operating characteristic (ROC) curve.
 Results: There were significant differences (both P<0.01) of the 2-dimension radiomics features between the two groups. The best performance (area under the ROC curve) of the established radiomics model was 0.971, with sensitivity and specificity at 85.7% and 94.1%, respectively.
 Conclusion: The performance of the machine learning-based radiomics model derived from CMR cine sequence for assessing the severity of mitral regurgitation was excellent, which can facilitate the computer-aided diagnosis and treatment in the era of artificial intelligence.
Heart ; Humans ; Magnetic Resonance Imaging ; Mitral Valve Insufficiency ; diagnostic imaging ; Reproducibility of Results ; Retrospective Studies

Objective To compare the cognitive function of patients with carotid stenosis combined with white matter lesions (WML)after carotid artery stenting (CAS). Methods Total 166 patients with carotid artery stenosis were collected.According to MRI imaging,30 patients with no white matter lesions were included in the control group and 136 patients with white matter lesions were included in the white matter lesions group. They were treated with carotid artery stenting and underwent evaluation on the safety and efficacy of perioperative surgery. CAS failed in two patients because of the inability of guidewire crossing in WML group. Two patients died after CAS (one for cardiac death and one for traumatic accident) in WML group. 162 patients received 1 year follow-up. Cognitive function was assessed before and after CAS. Results Before CAS,WML group's MMSE, digit span forward/backward test, verbal fluency test and MoCA scores (21.8±3.3、6.3±2.1、4.1±1.0、15.1±3.6、20.6±3.1) were lower compared with control (24.3±3.9、7.3±2.6、4.7±1.8、17.7±5.2、22.7±4.2) and ADAS-Cog score was higher compared with control ((15.1±3.3) vs.(12.7±3.3)), P=0.000、0.026、0.039、0.012、0.000、0.011.Three months after CAS,the MMSE,digit span forward test and MoCA scores (23.7±3.6,7.5± 2.4, 23.1±6.9) was higher significantly than those before treatment (21.8±4.3, 6.3±2.09, 20.6±4.13), P<0.05.And the scores of ADAS-Cog was lower((13.2±4.)vs.(15.1±4.3),P<0.05).The scores of digit span backward test in 6 months after treatment was significant higher than those before treatment (4.9 ±2.8,4.1 ±2.2,P<0.05). After 1 year of CAS, the improvement in scores of MMSE, digit span forward test, ADAS-Cog and MoCA in patients with carotid stenosis complicated with WML(3.5±1.3,1.6±0.6,-2.6±0.8,3.6±1.1)was higher significant than control(2.7±1.8, 1.2±0.8, -2.0± 1.3, 2.7 ±1.5),P<0.05. Conclusion CAS can improve cognitive function in Patients with carotid artery stenosis complicated with WML than those who without WML.

Objective To evaluate the effect of zinc deficiency factor on cognitive function after isoflurane anesthesia in mice with Alzheimer's disease (AD).Methods One hundred and forty-four APP/ PS1 transgenic mice with AD,weighing 22-28 g,aged 8-10 months,were divided into 6 groups (n=24 each) using a random number table:zinc adequate group (group ZA),zinc adequate plus isoflurane anesthesia group (group ZA+Iso),zinc deficiency group (group ZD),zinc deficiency plus isoflurane anesthesia group (group ZD+Iso),zinc treatment group (group ZT) and zinc treatment plus isoflurane anesthesia group (group ZT+Iso).The mice were fed a diet adequate in zinc and deionized water for 2 months in ZA and ZA+Iso groups.The mice were fed a diet low in zinc (0.01‰ zinc) and deionized water for 1 month in ZD and ZD+Iso groups.The mice were fed a diet adequate in zinc and 0.12‰ ZnSO4 · 7H2O water for 2 months in ZT and ZT+Iso groups.The mice underwent 2 h of anesthesia with 1.4％ isoflurane starting from the end of feeding in ZA+Iso,ZD+Iso and ZT+Iso groups.At 24 h after anesthesia,the mice were sacrificed and hippocampal tissues were obtained for determination of the contents of soluble amyloid beta protein 40 (Aβ40) and Aβ42 and insoluble Aβ40 and Aβ42 (using enzyme-linked immunosorbent assay) and expression of total Aβ,Aβ40,Aβ42,tau pSer396,tau pSer262 and tau pThr231 (by Western blot).Morris water maze test was performed at 24 h after anesthesia.Results There was no significant difference in each parameter between group ZA and group ZA+Iso and between group ZT and group ZT+Iso (P＞0.05).Compared with group ZD or group ZT+Iso,the escape latency was significantly prolonged,the space exploration time was shortened,the expression of hippocampal Aβ42,tau pSer396,tau pSer262 and tau pThr231 was up-regulated,and the contents of soluble and insoluble Aβ42 were increased in group ZD+Iso (P＜0.05 or 0.01).Conclusion Zinc deficiency can aggravate the impairment of cognitive function after isoflurane anesthesia in mice with AD,and the mechanism is related to the promotion of hippocampal Aβ aggregation and tau protein phosphorylation.

Objective To compare the degree of cell injury induced by 3D protein (SDLY11 and SDLY107) of enterovirus 71 (EV71) strains.Methods EV71 strains SDLY11 and SDLY107 were respectively isolated from children with mild and severe hand foot mouth disease.The target genes 11-3D-Flag and 107-3D-Flag were amplified by reverse transcription-polymerase chain reation (RT-PCR) and inserted into the eukaryotic expression vector pcDNA3.1.The recombinant plasmids 11-3D-Flag-pcDNA3.1 and 107-3D-Flag-pcDNA3.1 were transformed into Escherichia.coli DH5α, respectively, and were identified by enzyme digestion and sequencing.The recombinant plasmids were transfected into rhabdomyosarcoma (RD) cells, respectively.Expression of 3D protein was detected by indirect immunofluorescence assay and western blot.Cell injury induced by 3D protein was detected with lactate dehydrogenase (LDH) test, cell proliferation was detected by 3-(4,5-dimethylthiazolyl-2)-2,5-diphenylthiazolium bromide (MTT) test, and cell apoptosis was detected with Annexin-V and PI.Multiple comparisons among groups were analyzed using LSD-t test if multiple sets of variables were consistent with homogeneity of variance.If not, Dunnett T3 test was used.Results The 1 400 bp fragments were amplified by reverse tramscription (RT)-polymerase chain reaction (PCR), and the recombinant plasmids were digested by enzyme and the 1 400 bp and 5 400 bp fragments were obtained and identified.Gene sequencing showed that the sequences were consistent with the target genes.The specific fluorescence was observed by indirect immunofluorescence assay, and the western blot showed that the molecular weight of the target protein was 55×103.The LDH test showed that the A490 of SDLY11 3D protein transfection group (0.790±0.048) was higher than that of SDLY107 3D protein transfection group (0.641±0.018).The difference was statistically significant (t=5.14, P<0.05).The cell membrane damage caused by SDLY11 3D protein was more severe than SDLY107 3D protein.The MTT test showed that the A570 of SDLY11 3D protein transfection group (1.028±0.020) was lower than that of SDLY107 3D protein transfection group (1.081±0.002), and the difference was statistically significant (t=3.31, P<0.05).The effect on cell proliferation activity of SDLY11 3D protein was greater than SDLY107 3D protein.The results of Annexin-V/PI showed that the percentage of apoptotic cells of SDLY11 3D protein transfection group and SDLY107 3D protein transfection group were (1.471±0.246)% and (1.465±0.237)%, respectively, and the difference was not statistically significant (t=0.04, P=0.973).Conclusions Compared with the SDLY11 3D protein, SDLY107 3D protein induces slighter cell injury, has weaker effect on cell proliferation activity, and is more favorable for virus replication in cells.