BACKGROUND:Human periodontal ligament stem cells are potential functional cells for periodontal tissue engineering.However,long-term in vitro culture may lead to reduced stemness and replicative senescence of periodontal ligament stem cells,which may impair the therapeutic effect of human periodontal ligament stem cells. OBJECTIVE:To investigate the effect of oxymatrine on the stemness maintenance and osteogenic differentiation of periodontal ligament stem cells in vitro,and to explore the potential mechanism. METHODS:Periodontal ligament stem cells were isolated from human periodontal ligament tissues by tissue explant enzyme digestion and cultured.The surface markers of mesenchymal cells were identified by flow cytometry.Periodontal ligament stem cells were incubated with 0,2.5,5,and 10 μg/mL oxymatrine.The effect of oxymatrine on the proliferation activity of periodontal ligament stem cells was detected by CCK8 assay.The appropriate drug concentration for subsequent experiments was screened.Western blot assay was used to detect the expression of stem cell non-specific proteins SOX2 and OCT4 in periodontal ligament stem cells.qRT-PCR and western blot assay were used to detect the expression levels of related osteogenic genes and proteins in periodontal ligament stem cells. RESULTS AND CONCLUSION:(1)The results of CCK8 assay showed that 2.5 μg/mL oxymatrine significantly enhanced the proliferative activity of periodontal stem cells,and the subsequent experiment selected 2.5 μg/mL oxymatrine to intervene.(2)Compared with the blank control group,the protein expression level of SOX2,a stem marker of periodontal ligament stem cells in the oxymatrine group did not change significantly(P>0.05),and the expression of OCT4 was significantly up-regulated(P<0.05).(3)Compared with the osteogenic induction group,the osteogenic genes ALP,RUNX2 mRNA expression and their osteogenic associated protein ALP protein expression of periodontal ligament stem cells were significantly down-regulated in the oxymatrine+osteogenic induction group(P<0.05).(4)The oxymatrine up-regulated the expression of stemness markers of periodontal ligament stem cells and inhibited the bone differentiation of periodontal ligament stem cells,and the results of high-throughput sequencing showed that it may be associated with WNT2,WNT16,COMP,and BMP6.

Objective:To study the predictive values of lung ultrasound (LUS) score and Downes score in selecting respiratory support strategies for newborns with dyspnea.Methods:From September 2021 to July 2022, newborns admitted to our hospital with dyspnea were selected and assigned into the non-invasive respiratory support (N) group, invasive respiratory support (I) group and control (C) group based on the respiratory support strategies on admission. LUS scores and Downes scores at 6, 24, and 48 h after birth were recorded. ROC curves were drawn to determine the predictive values of LUS and Downes scores for respiratory support strategies.Results:A total of 263 cases were enrolled, including 105 cases in N group, 56 cases in I group and 102 cases in C group. The differences of LUS and Downes scores between the three groups at the same timepoint were statistically significant with I group had the highest scores, N group second and C group lowest ( P<0.05). LUS and Downes scores within each group at different timepoints were significantly different ( P<0.05).In all three groups, LUS and Downes scores were decreased with longer duration of treatment. LUS score, Downes score and PaO 2/FiO 2 were positively correlated with each other ( P<0.05). The area under the curve (AUC) of LUS score and Downes score predicting non-invasive respiratory support within 6 h after birth were 0.900 (95% CI 0.861-0.940, P<0.05) and 0.889 (95% CI 0.847-0.931, P<0.05), respectively, with the same cutoff of 2.5. The AUC of the combination of LUS and Downes scores predicting non-invasive respiratory support was 0.944 (95% CI 0.915-0.973, P<0.05). The AUC of LUS score and Downes score predicting invasive respiratory support were 0.979 (95% CI 0.963-0.995, P<0.05) and 0.831 (95% CI 0.760-0.902, P<0.05), respectively, with the same cutoff of 5.5. The AUC of the combination of LUS and Downes scores predicting invasive respiratory support was 0.985 (95% CI 0.972-0.998, P<0.05). Conclusions:Both LUS score and Downes score have certain predictive values for respiratory support strategies in newborns with dyspnea.

Objective:To investigate the status of hypoglycemia fear, diabetes distress, self-regulatory fatigue and self-management in type 2 diabetes patients. The chain mediating effects of self-regulated fatigue and psychological distress on hypoglycemic fear and self-management were investigated.Methods:The Hypoglycemia Fear Survey-Worry Subscale (CHFSII-WS), Diabetes Distress Scale (DDS), Self-Regulatory Fatigue Scale (SRF-S) and Summary of Diabetes Self-Care Activities (SDSCA) were used to investigate the type 2 diabetes patients from the department of Endocrinology of the First Affiliated Hospital in Jinzhou Medical University. And constructed the structural equation model.Results:The scores of fear of hypoglycemia in type 2 diabetes patients were positively correlated with psychological pain and self-regulating fatigue ( r=0.739, 0.625, P<0.05), but negatively correlated with self-management level ( r=-0.602, P<0.05). The psychological pain score was positively correlated with the self-regulating fatigue score ( r=0.669, P<0.05) and negatively correlated with the self-management level score ( r=-0.609, P<0.05). The score of self-regulation fatigue was negatively correlated with the score of self-management ( r=-0.596, P<0.05). Pathway analysis showed that hypoglycemia fear could directly affect self-management behavior, indirectly predict self-management level through self-regulation fatigue and psychological pain respectively, and negatively affect self-management behavior through chain mediation of self-regulation fatigue and psychological pain ( χ2/ df=3.079, GFI=0.920, CFI=0.961, NFI=0.943, IFI=0.961, RMSEA=0.078). Conclusions:The Self-regulated fatigue and psychological distress acts as the chain mediators of hypoglycemic fear and self-management in patients with type 2 diabetes.

Objective:To study the clinical characteristics of children with adenovirus pneumonia complicated with liver injury and the understanding of clinicians.Methods:The children diagnosed with community-acquired adenovirus pneumonia were enrolled from January 1, 2013 to December 31, 2020. The clinical characteristics of the normal liver function group and the liver injured group were compared, and multivariate regression was applied.Results:Among the 3 294 enrolled children, 1 704 children had elevated alanine aminotransferase and/or aspartate aminotransferase on admission, and the liver function impairment rate was 51.7% (1 704/3 294), of which 1 671 cases were mildly injured (98.06%, 1 671/1 704), 28 cases were moderately injured (1.64%, 28/1 704), and 5 cases were severely injured (0.29%, 5/1 704). Compared with the normal liver function group, the proportion of male children in the liver injured group was higher (68.1% vs. 64.2%, P=0.017), and the age was younger[16(9, 30) months vs. 38(19, 53) months, P<0.001]. Children with liver injury were more likely to have wheezing (32.9% vs. 18.7%, P<0.001), shortness of breath (13.7% vs. 9.1%, P<0.001), and diarrhea (11.0% vs. 4.7%, P<0.001). The white blood cell counts, neutrophil counts and hemoglobin in the liver injured group were lower than those in the normal group, while the lymphocyte counts were higher than those in the normal group. The levels of creatine kinase, cardiac creatine kinase and lactate dehydrogenase were higher. The proportion of severe pneumonia in the liver injured group (26.9% vs. 11.2%, P<0.001) was higher than that in the normal liver function group, and the case fatality rate was significantly higher (1.47% vs. 0.25%, P<0.001). Comparing the probability of liver injury in children with mild and severe adenovirus pneumonia, it was found that the severe pneumonia group had more frequent liver injury (73.0% vs 46.6%, P<0.001), and both moderate and severe liver injury groups were higher than in the mild group. Further multivariate logistic regression analysis did not find a significant factor associated with the incidence of liver injury. Conclusions:The incidence of adenovirus pneumonia in children with liver injury was high, and mild injury is more common. Children with liver injury were younger, had more severe clinical symptoms such as wheezing and shortness of breath, and higher proportion of severe pneumonia and mortality. Among them, children with severe pneumonia had a higher proportion of liver injury, and moderate to severe injury was more common. The monitoring of children with different degrees of liver injury should be strengthened.

This paper reports 3 cases of acute hepatitis of unknown cause in children who met the case definition of WHO. Human adenovirus group C was detected in case 3, and the quantity of viral DNA was relatively high, which may be related to the liver function damage in the patient, but its role in pathogenesis needs further study to confirm.

Objective:To investigate the distribution and clinical significance of subtypes of antimitochondrial antibodies (AMA)-M2, M4, M9 in primary biliary cholangitis (PBC).Methods:A total of 1 367 patients were detected with AMA-M2, M4, M9 in Peking Union Medical College Hospital (PUMCH) from Jan 2014 to Dec 2019 and the clinical parameters were collected. The distribution patterns of AMA subtypes in different groups were analyzed and the diagnostic sensitivity and specificity of AMA subtypes in PBC were calculated. Chi-square test was used for statistical analysis.Results:In 1 367 patients, 236 of whom were positive for AMA subtypes. The positivity of AMA subtypes in female was significantly higher than in male (20.34% vs 9.41%, χ2=23.792, P<0.01). In addition, the positivity of AMA subtypes was significantly higher in 30-65 years old patients than in patients younger than 30 years old or older than 65 years old [(20.00%(193/965) vs 10.97%(17/155) vs 10.53%(26/247), χ2=17.209, P<0.01]. 110 patients with positive AMA subtypes were diagnosed with PBC. The diagnostic sensitivity and specificity of AMA-M2 were both desirable [94.64%(106/112) and 92.35%(1 159/1 255)]. Although the specificity of AMA-M4 was as high as 99.12%(1 244/1 255), its sensitivity was very low [15.18%(17/122)]. Combined detection of different AMA subtypes could not improve the diagnostic sensitivity and specificity significantly. Diseases other than PBC can be positive for AMA subtypes, predominantly for AMA-M2. Conclusion:Female and 30-65 years old patients were more frequently positive for AMA subtypes. AMA-M2 was the most valuable AMA subtype for diagnosing PBC.

OBJECTIVE：To establish an overall quality evaluation model for Agrimonia pilosa based on extract and characteristic spectrum，and to provide evidence for comprehensive quality evaluation of the medicinal material and screening of high-quality provenance. METHODS ：Referring to different extraction method and solvent condition stated in 2015 edition of Chinese Pharmacopoeia （part Ⅳ），using the content and total peak area of HPLC characteristic chromatogram of extract as indexes，and the extraction technology was optimized by weighted comprehensive score. HPLC characteristic spectrum of 15 batches of A. pilosa was established ，and similarity evaluation and characteristic peak identification were performed. SPSS 25.0 software was used to conduct single factor analysis and Pearson correlation analysis for the extract content and total peak area of A. pilosa from different origins. The quality of medicinal materials from different origins were compared. Entropy weight TOPSIS method was adopted to evaluate comprehensive quality of A. pilosa using the extract content and total peak area of 15 batches of A. pilosa . RESULTS ：The extraction technology of A. pilosa extract，which was extracted with hot dip plating using 50％ ethanol as solvent ，was optimized. The similarity of 15 batches of A. pilosa was higher than 0.92，and 4 characteristic components were identified（ellagic acid ，quercetin，apigenin，kaempferol）. There were significant differences in average extract content and total peak area of characteristic chromatogram of A. pilosa from different origins （P＜0.05 or P＜0.01），and there was a certain positive correlation between them （r＝0.86，P＜0.01）. Results of entropy weight TOPSIS evaluation showed that the average Ci values of A. pilosa in Anhui ，Zhejiang，Sichuan，Henan and Jiangsu provinces were 0.689，0.351，0.218，0.308 and 0.361 respectively. CONCLUSIONS：The quality of A. pilosa from Anhui was the best ，that from Zhejiang and Jiangsu was better ，that from Henan was the second ，and that from Sichuan was poor. Established extraction technology and characteristic spectrum determination method of A. pilosa are stable and feasible. The entropy weight TOPSIS model is objective and quantifiable for comprehensive quality evaluation of A. pilosa ，and can effectively evaluate its quality.

Objective: The gene engineering technique was used to express the P[6] genotype rotavirus (rotaviruses, RVs) GST-VP8^*-Z84 protein from pigs, and the binding characteristics of the protein to oligosaccharide and salivary receptor were studied. Methods: The GST-VP8^*-Z84 protein was purified by GST Escherichia coli expression system and affinity chromatography using porcine P[6]. Enzyme-linked immunosorbent assay (ELISA) saliva binding test and oligosaccharide binding test were used to analyze the binding characteristics of the genotype to receptors. Results: Porcine P[6] GST-VP8^*-Z84 protein bound well to mucin core 2. Conclusions The potential receptor of P[6] RV may be the core of mucin, which may provide the experimental basis and theoretical basis for the mechanism of rotavirus and receptor interaction and the development of RV vaccine and highly effective therapeutic drugs.

Objective: The VP8^* core protein of rotavirus P[6] genotype LL4260 was purified by prokaryotic expression, which is important for further study of protein structure and function. Methods: The P[6] genotype LL4260 strain was obtained by PCR.The recombinant plasmid pET30 a-LL4260VP8^*core was inserted into pET30 a vector and transformed into BL21 (DE3) competent cells with the correct recombinant plasmid. The expressed protein is purified by affinity chromatography and molecular sieve chromatography. Results: The pET30 a-LL4260VP8^* core region protein is soluble in the supernatant and proteins of approximately 22 kDa are identified by electrophoresis using purified proteins. Conclusions In this study, LL4260 containing pET30 a-LL4260VP8^* core plasmid was successfully constructed and LL4260 strain VP8^* protein was expressed.

Objective To explore the clinical treatment of patients with stage Ⅲ carcinoma of colon and rectum by tegafur chemotherapeutic drugs combined with standardized adjuvant therapy. Methods The data of 60 cases of carcinoma of colon and rectum treated in Wenzhou Central hospital from January 2013 to December 2015 were collected. The patients were divided into three groups, non standardized adjuvant treatment group of 18 cases, standardized adjuvant treatment group of 15 cases and tegafur drug combined with standardized treatment group of 27 cases. Recorded and compared the survival rate of the three groups within three years, the statistics of each group of disease-free survival (DFS) and overall survival rate (OS). According to the duration of treatment group, the use of oxaliplatin combined with 5-fluorouracil and leucovorin (FOLFOX) treatment, analyzed and recorded OS and DFS. Follow-up examinations were performed in patients who did not receive adjuvant therapy. Recorded and compared the OS and DFS of each group. Results Tegafur drug combined with standardized treatment group DFS and OS were significantly higher than the other two groups, the difference was statistically significant (P<0.05). In addition, when treated with FOLFOX regimen, the DFS and OS of group Ⅲ were significantly higher than those of groupⅠand groupⅡ, the difference was statistically significant (P<0.05). In the follow-up survey of three groups, the rates of recurrence and death in patients treated with tegafur drug combined with standardized treatment group were significantly lower than those in the non standardized adjuvant treatment group and the standardized adjuvant treatment group. Conclusion Tegafur combined with adjuvant therapy can significantly improve the standardization of the rate of carcinoma of colon and rectum and the rate of disease-free survival, it is suggested that doctors use chemotherapy combined with auxiliary standardized therapy, can significantly improve the survival of patients, and reduce the risk of disease recurrence rate.