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.

[Purpose]To investigate the trends of incidence and age at onset of uterine corpus can-cer in Jiangsu cancer registration areas from 2009 to 2019.[Methods]The continuous monitoring data of uterine corpus cancer from 2009 to 2019 were collected from 16 cancer registries in Jiang-su Province.The crude incidence rate,the age-standardized incidence rate by Chinese standard population(ASIRC),crude and adjusted mean age,and standardized age-specific incidence composition were calculated.The average annual percentage change(AAPC)were analyzed by the Joinpoint regression model.The linear regression model was used to analyze the relationship be-tween mean age at onset and year.The standardized age-specific incidence composition in 2009 and 2019 were compared.[Results]The ASIRC of uterine corpus cancer in all registration areas and in rural areas of Jiangsu Province showed upward trends with AAPC of 1.78％and 2.38％,re-spectively(P＜0.05),but not showed in the urban areas(AAPC=1.30％,P＞0.05).The crude mean age at onset increased from 56.48 years old in 2009 to 58.26 years old in 2019 with an average annual growth of 0.173 years old(P=0.001).After the population structure standardized,the trends disappeared in all registration areas.[Conclusion]From 2009 to 2019,the standardized incidence rates of uterine corpus cancer were on rise in Jiangsu cancer registration areas,especially in the age group of 50 to 59 years old.

Objective: To investigate the trends in the disease burden of leukemia in Jiangsu Province from 1990 to 2019, so as to provide insights into leukemia control in Jiangsu Province. Methods: The prevalence, incidence, death and disability adjusted life years (DALYs) of leukemia in Jiangsu Province from 1990 to 2019 were retrieved from the 2019 Global Burden of Disease Study (GBD 2019) database, and standardized by the age structure of Chinese populations in 2000. The trends in the disease burden of leukemia in Jiangsu Province from 1990 to 2019 were analyzed using average annual percent change (AAPC). Results : The average annual standardized prevalence, incidence, mortality and DALY rate of leukemia were 63.63/105, 9.76/105, 4.10/105 and 194.83/105 in Jiangsu Province from 1990 to 2019, respectively. The average annual standardized prevalence (AAPC=1.420%, t=5.644, P<0.001) and incidence (AAPC=0.806%, t=3.505, P<0.001) of leukemia appeared a tendency towards a rise, while the average annual standardized mortality (AAPC=-1.589%, t=-14.714, P<0.001) and DALY rate (AAPC=-1.849%, t=-9.046, P<0.001) of leukemia appeared a tendency towards a decline in Jiangsu Province from 1990 to 2019. Higher average annual standardized prevalence (65.27/105 vs. 62.38/105, P<0.001), incidence (10.32/105 vs. 9.29/105, P<0.001), mortality (4.69/105 vs. 3.57/105, P<0.001) and DALY rate of leukemia (216.94/105 vs. 172.80/105, P<0.001) were estimated among men than among women. The crude prevalence of leukemia peaked among patients at ages of 0 to 14 years and 60 to 74 years, and the crude incidence, DALY rate and mortality of leukemia peaked at ages of 0 to 14 years and 75 years and older. Conclusions The mortality and DALY of leukemia appeared a tendency towards a decline and the prevalence and incidence appeared a tendency towards a rise in Jiangsu Province from 1990 to 2019; however, there were high disease burdens of leukemia among men, children and the elderly.

Objective:To express prokaryotically GII.4 human norovirus (HuNoV) VP2 protein and to prepare polyclonal antibody against VP2.Methods:Design specific primers to amplify the VP2 gene of GII.4 HuNoV, digest and connect to the prokaryotic expression vector pGEX-6P-1, transform the correctly identified recombinant plasmid into BL21 ( DE3) competent cells.Pick out and shake the monoclonal bacteria, and add IPTG to induce recombinant GST-VP2. The fusion protein was expressed, purified by GST affinity chromatography and digested to obtain GII.4 HuNoV VP2 protein. The relative molecular mass (Mr.×10 3) of the purified HuNoV VP2 protein was analyzed by SDS-PAGE. BALB/c mice were immunized with purified GII.4 HuNoV VP2 protein (0.5 mg/ml) to prepare polyclonal antibodies. Results:The VP2 protein of GII.4 HuNoV was successfully expressed and purified, with a relative molecular mass (Mr.×10 3) of about 29; the VP2 polyclonal antibody of GII.4 HuNoV was successfully prepared and its titer was as high as 1∶1 280 000. Western blot and indirect ELISA analysis showed that the polyclonal antibody could specifically bind to the VP2 antigen of GII.4 HuNoV. Conclusions:The purified GII.4 HuNoV VP2 after prokaryotic fusion expression can be used to prepare high titer polyclonal antibody.

Objective:To understand the prevalence of human parechovirus (HPeV) in neonates of Hunan Provincial People’s Hospital, and analyze genetic evolutionary characteristics.Methods:From June to September 2021, fecal samples of inpatient neonates were collected in Hunan Provincial People′s Hospital. TaqMan real-time qPCR and RT-PCR were used for HPeV screening and genotyping. High-throughput sequencing and PCR were used to obtain whole genomes. Phylogenetic analysis was performed after sequencing.Results:A total of 123 fecal samples of neonates were collected, of which 22 were HPeV positive, with 17.89% positive rate. All the strains belonged to the HPeV-1 genotype. One full-length genomic sequence of 7 269 bp were obtained, and provisionally named Hunan/HPeV/2021, which has the highest nucleotide identity with known HPeV-1 genotype, with 86.6%-91.9% nucleotide identity. The nucleotide and amino acid identity of open reading frame (ORF) with known similar sequences were 90.3%-92.6% and 97.3%-98.3%, respectively. The phylogenetic analysis showed that Hunan/HPeV/2021 belongs to the HPeV-1 genotype, which is clustered into the same clade as the popular HPeV-1 strains in China.Conclusions:HPeV has a high prevalence in inpatient neonates of Hunan Provincial People’s Hospital and belong to the HPeV-1 genotype.

Objective: To investigate the incidence and trends in incidence of stroke among residents in Lianyungang City, Jiangsu Province from 2014 to 2020, so as to provide insights into stroke control. Methods: The stroke morbidity data in Lianyungang City from 2014 to 2020 were captured from Lianyungang Municipal System for Monitoring of Cardiovascular and Cerebrovascular Diseases, and standardized by the sixth national population census data in China in 2010. The trends in stroke incidence were evaluated using average annual percentage change (AAPC), and the gender-, age-, region- and disease type-specific incidence and trends in incidence of stroke were descriptively analyzed. Results: Totally 162 423 new stroke cases were diagnosed in Lianyungang City from 2014 to 2020 new case, and the annual mean crude and standardized incidence rates were 435.32/105 and 357.28/105, which both appeared a tendency towards a rise (AAPC=12.61%, t=39.098, P<0.001; AAPC=9.91%, t=6.123, P<0.001) . The standardized incidence of stroke was significantly higher in men than in women (389.63/105 vs. 327.17/105; χ2=4.056, P=0.044). The crude incidence of stroke appeared a tendency towards a rise with age among residents (χ2trend=258.219, P<0.001), and appeared a tendency towards a rise among residents at ages of <35 (AAPC=22.52%, t=6.723, P<0.001), 35 to 64 (AAPC=9.78%, t=7.867, P<0.001) and 65 years and older (AAPC=10.24%, t=5.156, P<0.001). The crude incidence was higher in rural areas than in urban areas (437.72/105 vs. 425.31/105; χ2=20.709, P<0.001). Ischemic stroke was the predominant type (139 680 cases, 86.00% of all cases), and both the crude and standardized incidence appeared a tendency towards a rise (AAPC=13.79%, t=16.865, P<0.001; AAPC=10.88%, t=5.128, P<0.001). Conclusion The incidence of stroke appeared a tendency towards a rise among residents living in Lianyungang City from 2014 to 2020. Men, middle-age and elderly residents and rural residents are high-risk populations for stroke control.

Objective To evaluate the safety and efficacy of irreversible electroporation (IRE) combined with neoadjuvant chemotherapy in patients with locally advanced pancreatic cancer. Methods We searched PubMed, Embase, Cochrane Library, Web of Science, China Biomedical Literature Database, CNKI, Wanfang, and VIP databases for articles dated from the establishment of each database to March 2022. Meta-analysis was performed using RevMan5.4 software. Results A total of 3970 patients with locally advanced pancreatic cancer were enrolled in eight studies, including one randomized controlled trial, four retrospective studies, and three prospective studies. The patients were divided into the combined therapy group with 344 patients and the chemotherapy-only group with 3626 patients. Meta-analysis showed that the overall survival of patients in the combined therapy group was significantly higher than that in the chemotherapy-only group (OR=4.52; 95%CI: 2.63-7.77; P < 0.00001). However, no significant difference existed in the disease control rate between the combined therapy group and the chemotherapy-only group (OR=0.58; 95%CI: 0.02-18.74; P=0.76). Moreover, no significant difference existed in the disease progression between the two groups (OR=0.49; 95%CI: 0.23-1.02; P=0.06). The combination of neoadjuvant chemotherapy and IRE had no significant effect on the incidence of adverse reactions of gastrointestinal reaction (OR=0.37; 95%CI: 0.10-1.34; P=0.13) and bone marrow suppression (OR=0.61; 95%CI: 0.26-1.40; P=0.24). Conclusion IRE combined with neoadjuvant chemotherapy can remarkably improve the prognosis of patients with locally advanced pancreatic cancer, and significantly prolong the overall survival.

Objective:To optimize and evaluate the pretreatment method for high-throughput sequencing of fecal and tissue samples to improve the sensitivity of high-throughput sequencing in the study of virome.Methods:For fecal samples, five virus-positive samples that have been detoxified from feces were selected, mixed as the simulated samples, and filters made of different materials were used, different processing times were set for nucleases, and different kits were used to extract nucleic acid. For tissue samples, two virus-positive samples that have been detected in animal tissues, filter and nuclease treatment were used, and different extraction method were used to extract nucleic acid. TaqMan real-time PCR was used to quantitatively evaluate the impact of each treatment on the virus and the advantages and disadvantages were compared. We used the SYBR Green real-time PCR quantitative method to evaluate the removal effect of the above method on bacteria 16S rRNA and host 12S rRNA genome.Results:For fecal samples, the 0.22 μm PES filter showed a better filtering effect, and the PVDF material filter reduces the sample volume; 2 h nuclease digestion was better than 1 h digestion to remove bacteria, and the virus loss was less; the use of RPMK kits can effectively reduce bacteria, but the effect of extracting some viruses was poor, and the MVSK kit has a better effect of extracting viral nucleic acid. For tissue samples, 0.22 μm PES filter filtration, nuclease digestion for 1 h and VNAEK II kit extraction of nucleic acid were the best, Trizol LS combined with the RPMK method was better for gDNA removal, but the virus loss was larger. The virus loss of the whole process of the pretreatment method of feces and tissue samples was (1.7-3.0) Ct and (1.6-2.5) Ct, respectively.Conclusions:The optimal method for fecal samples was to filter with a PES filter, then digest with nuclease for 2 hours, and then extract nucleic acids using the MVSK kit; the optimal method for tissue samples was to filter with a PES filter, then perform 1 h nuclease digestion, and then use VNAEK II kit to extract nucleic acids.

ObjectiveTo investigate the correlation between systemic immune-inflammation index (SII) and prognosis in patients with hepatic alveolar echinococcosis. MethodsA retrospective analysis was performed for the clinical data of 242 patients who were admitted to Department of Hepatopancreatobiliary Surgery, Qinghai University Affiliated Hospital, from January 2015 to December 2018 and underwent surgery for hepatic alveolar echinococcosis, and SII was calculated. The chi-square test was used for comparison of categorical data between two groups, and a Spearman correlation analysis was performed. The receiver operating characteristic (ROC) curve was used to determine the optimal cut-off value of SII; the Kaplan-Meier method was used to plot survival curves and analyze overall survival time in the two groups, and the log-rank test was used for comparison of survival rates between the two groups; univariate and multivariate Cox regression analyses were used to identify the influencing factors for the prognosis of patients with hepatic alveolar echinococcosis. ResultsThe Spearman correlation analysis showed that SII was positively correlated with the postoperative fatality rate of patients with hepatic alveolar echinococcosis (r=0.267, P＜0.001). The ROC curve showed that the optimal cut-off value of SII before surgery was 758.92, and based on this, 242 patients with hepatic alveolar echinococcosis were divided into low SII (SII ≤758.92) group with 126 patients and high SII (SII ＞758.92) group with 116 patients. The low SII group had 1-, 3-, and 5-year survival rates of 98.20%, 88.47%, and 6610%, respectively, and the high SII group had 1-, 3-, and 5-year survival rates of 90.80%, 53.05%, and 27.40%, respectively. The low SII group had a cumulative survival rate of ＞50% and a mean survival time of 55.584 months (95% confidence interval［CI］: 53550-57.617), while the high SII group had a cumulative survival rate of ＜50%, a mean survival time of 39.384 months (95% CI: 35.070-43.698), and a median survival time of 43 months (95% CI: 34.694-51.306). The low SII group had a significantly better survival rate than the high SII group, and there was a significant difference in overall survival rate between the two groups (χ2=46.979, P＜005). The univariate analysis showed that SII ＞758.92 (hazard ratio ［HR］=5.907, 95% CI: 3.386-10.306, P=0.001) was an influencing factor for the overall survival time of patients with hepatic alveolar echinococcosis, and the multivariate Cox regression analysis showed that preoperative peripheral blood SII (HR=3.507, 95% CI: 1.911-6.435, P=0.001) was an independent risk factor for the overall survival rate of patients with hepatic alveolar echinococcosis. ConclusionPreoperative SII level is clearly correlated with the prognosis of patients with hepatic alveolar echinococcosis and can thus be used as a clinical indicator to evaluate the prognosis of patients. The higher the peripheral blood SII before surgery, the worse the prognosis of patients.