Objective:To explore the role and mechanism of neurotrophin-3(NT-3)in promoting neurological func-tion recovery after cerebral ischemia-reperfusion injury in rats.Methods:Twenty-four SD rats were randomly divided into sham operation group(Sham),middle cerebral artery occlusion/reperfusion(MCAO/R)group,and MCAO/R+NT-3 group.The neurological function scores of rats in each group were evaluated using the modified Garcia score.Western Blot was used to detect the expression of NT-3 and LC3B in brain tissues of rats.Culture dishes with the same density of neurons were randomly divided into normal group(Normal),oxygen-glucose deprivation(OGD)group,OGD+NT-3 group,OGD+NT-3+PF-06273340(TrkC inhibitor)group,OGD+NT-3+ZSTK474(PI3K inhibitor)group,and OGD+NT-3+CCT128930(AKT inhibitor)group.Western Blot was used to detect the expression of TrkC,the phosphorylation of PI3K/AKT,and LC3B in neurons.The morphological changes of neurons and the phenomenon of neuronal autophagy were observed using autophagy-specific fluorescent staining.Results:The animal experiment found that the expression of NT-3 increased in the brain tissue with ischemia-reperfusion injury(P＜0.05),and after the treatment with exogenous NT-3,the modified Garcia score increased(P＜0.05),and the level of autophagy weakened(P＜0.05).The cell experiment found that NT-3 can inhibit neuronal autophagy under ischemic hypoxia and maintain the neuronal morphology to the maximum extent.After using PF-06273340,the expression of p-PI3K and p-AKT de-creased(P＜0.05).After using ZSTK474 and CCT128930,the autophagy-inhibiting effect of NT-3 weakened(P＜0.05).Conclusion:NT-3 inhibits autophagy via the PI3K/AKT signaling pathway to maintain neuronal survival,thereby promoting the recovery of neurological function after cerebral ischemia-reperfusion injury in rats.

BACKGROUND:The research of dental stem cells in the fields of regenerative medicine and tissue engineering has been deepening,bringing hope for the repair of tooth-related tissues and the treatment of systemic diseases.However,there is a lack of systematic research and analysis on the biological characteristics of dental stem cells in different age groups. OBJECTIVE:To explore the biological characteristics of the human deciduous tooth and permanent tooth pulp stem cells cultured in umbilical cord blood platelet lysate to provide a reliable basis for human platelet lysates to replace fetal bovine serum. METHODS:The pulp tissues of deciduous teeth,juvenile permanent teeth and adult permanent teeth were taken out and cultured in DMEM/F-12 medium supplemented with 10%fetal bovine serum or different concentrations(5%,10%and 15%)of human platelet lysates.Cell proliferation in the four groups was detected by cytometry.The optimal concentration of human platelet lysates was selected for subsequent experiments.Under the optimal concentration of human platelet lysates,human deciduous tooth and juvenile and adult permanent tooth pulp stem cells were cultured in vitro.The cell growth status was observed under the microscope.The specific antigen on the cell surface was detected by flow cytometry.The cell proliferation ability was tested by the cell counting method and CCK-8 assay.The cell differentiation ability in vitro was observed by a three-line differentiation assay. RESULTS AND CONCLUSION:(1)The cell proliferation rate of the 10%human platelet lysate group was the highest.(2)In all three groups,fusiform fibrous cells grew and expanded from around the tissue block.There was no significant difference between deciduous teeth and juvenile permanent tooth cells,but the adult permanent tooth cells were larger than the deciduous and juvenile permanent tooth cells of the same generation.(3)The results of flow cytometry showed that deciduous teeth,juvenile permanent teeth and adult permanent teeth conformed to the phenotypic characteristics of mesenchymal stem cells.(4)The proliferative capacity of adult permanent dental pulp stem cells was significantly lower than those of deciduous teeth and juvenile permanent dental pulp stem cells(P<0.01).(5)mRNA expressions of osteoblast-related genes alkaline phosphatase and bone morphogenetic protein 2,lipoprotein lipase and peroxisome proliferator-activated receptor γ2,mRNA expressions of chondroblast related gene type II collagen α1 and cartilage oligomeric matrix protein in adult pulp stem cells of permanent teeth were significantly lower than those of deciduous teeth and juvenile permanent teeth pulp stem cells(P<0.01).(6)Compared with adult dental pulp stem cells,human deciduous teeth and juvenile permanent teeth dental pulp stem cells have the stronger proliferative capacity and multidirectional differentiation potential,and are more suitable for clinical research and disease treatment.

Objective:To investigate dietary patterns of individuals aged ≥50 in Shanghai and analyze their association with frailty.Methods:Using data from the third wave of the Study on Global Ageing and Adult Health in Shanghai conducted between 2018 and 2019. We collected the frequency and average intake of food by the food frequency questionnaire. Factor analysis was used to extract dietary patterns, and a frailty index was constructed using the ratio of the cumulative total score of health deficits to 35 health-related variables considered. We used an ordinal multinomial logistic regression model to analyze the association between dietary patterns and frailty.Results:A total of 3 274 participants aged (67.9±9.2) years were included in the study, including 1 971 (60.2%) men and 1 303 (39.8%) women. We extracted four dietary patterns: high-protein-nuts pattern, potato-bean-vegetable-fruit pattern, poultry-meat pattern, and high-oil-salt pattern. After adjusting for confounding factors, the logistic regression analysis showed that compared with the high-oil-salt pattern, the high-protein-nuts pattern was negatively associated with the risk of higher frailty ( OR=0.743, 95% CI: 0.580-0.951). We did not find an association between dietary patterns and frailty between the different gender groups. In the age group 50-64, the high-protein-nuts and potato-bean-vegetable-fruit patterns were negatively correlated with a higher degree of frailty than the high-oil-salt pattern. In the low-level physical activity group, the high-protein-nuts pattern was negatively correlated with a higher degree of frailty than the high-oil-salt pattern ( OR=0.509, 95% CI: 0.361-0.720). However, we found no significant effect of the high-protein nuts pattern, potato-bean-vegetable-fruit pattern, and poultry-meat pattern on the risk of higher frailty compared to the high-oil-salt pattern in the moderate to high level of physical activity group. Conclusions:Compared to the high-oil-salt pattern, dietary patterns with a higher intake of high-protein nuts, potatoes, legumes, and fruits and vegetables might be associated with a lower risk of higher frailty in residents aged 50-64 years of age than with a high oil and salt pattern. At the same time, it may have a more significant protective effect in people with lower physical activity levels. It is suggested that a diet rich in high-protein foods, nuts, potatoes, beans, vegetables, and fruits may help reduce and delay the risk of frailty.

Objective:To understand the association between vitamin D level and grip strength in people aged ≥50 years in Shanghai.Methods:Data were obtained from the WHO's Study on Global Ageing and Adult Health in Shanghai during 2018-2019. Logistic regression model was used to analyze the association between vitamin D level and grip strength, and a stratified analysis was conducted for different gender, age and dairy product intake groups. Restricted cubic spline was used to evaluate the dose-response association between vitamin D level and low grip strength.Results:A total of 4 391 participants were included in the study, including 2 054 men (46.8%), with an average age of (67.02±8.81) years. And 1 421 individuals (32.4%) had low grip strength; 1 533 individuals (34.9%) had vitamin D deficiency, and 401 individuals (9.1%) had vitamin D deficiency. After adjusted for confounding factors, the logistic regression results analysis showed that individuals with vitamin D deficiency had a higher risk for low grip strength ( OR=1.41, 95% CI: 1.09-1.83). In men, after adjusting for confounding factors, vitamin D deficiency was positively associated with the risk for low grip strength ( OR=1.67, 95% CI: 1.12-2.50), but there was no significant association between vitamin D level and grip strength in women ( OR=1.30, 95% CI: 0.97-1.74). In age group 60-69 years and ≥80 years, there was significant association between vitamin D deficiency and low grip strength after adjusting for confounding factors ( OR=1.57, 95% CI: 1.05-2.35; OR=2.40, 95% CI: 1.08-5.31). In people who had daily intake of dairy product <250 ml, there was positive association between vitamin D deficiency and low grip strength, but there was no significant association in people who had daily dairy product ≥250 ml after adjusting for confounding factors. The restrictive cubic spline demonstrated that risk of low grip strength might decreased with the increase of vitamin D levels, however, the difference was not significant ( P>0.05). Conclusions:This study demonstrated that there is association between vitamin D level and grip strength. People with vitamin D deficiency have higher risk for low grip strength.

Objective:To investigate the clinical features and therapeutic efficacy of patients with hereditary leiomyomatosis and renal cell carcinoma(RCC) syndrome-associated RCC (HLRCC-RCC) in China.Methods:The clinical data of 119 HLRCC-RCC patients with fumarate hydratase (FH) germline mutation confirmed by genetic diagnosis from 15 medical centers nationwide from January 2008 to December 2021 were retrospectively analyzed. Among them, 73 were male and 46 were female. The median age was 38(13, 74) years. The median tumor diameter was 6.5 (1.0, 20.5) cm. There were 38 cases (31.9%) in stage Ⅰ-Ⅱand 81 cases (68.1%) in stage Ⅲ-Ⅳ. In this group, only 11 of 119 HLRCC-RCC patients presented with skin smooth muscle tumors, and 44 of 46 female HLRCC-RCC patients had a history of uterine fibroids. The pathological characteristics, treatment methods, prognosis and survival of the patients were summarized.Results:A total of 86 patients underwent surgical treatment, including 70 cases of radical nephrectomy, 5 cases of partial nephrectomy, and 11 cases of reductive nephrectomy. The other 33 patients with newly diagnosed metastasis underwent renal puncture biopsy. The results of genetic testing showed that 94 patients had FH gene point mutation, 18 had FH gene insertion/deletion mutation, 4 had FH gene splicing mutation, 2 had FH gene large fragment deletion and 1 had FH gene copy number mutation. Immunohistochemical staining showed strong 2-succinocysteine (2-SC) positive and FH negative in 113 patients. A total of 102 patients received systematic treatment, including 44 newly diagnosed patients with metastasis and 58 patients with postoperative metastasis. Among them, 33 patients were treated with tyrosine kinase inhibitor (TKI) combined with immune checkpoint inhibitor (ICI), 8 patients were treated with bevacizumab combined with erlotinib, and 61 patients were treated with TKI monotherapy. Survival analysis showed that the median progression-free survival (PFS) of TKI combined with ICI was 18 (5, 38) months, and the median overall survival (OS) was not reached. The median PFS and OS were 12 (5, 14) months and 30 (10, 32) months in the bevacizumab combined with erlotinib treatment group, respectively. The median PFS and OS were 10 (3, 64) months and 44 (10, 74) months in the TKI monotherapy group, respectively. PFS ( P=0.009) and OS ( P=0.006) in TKI combined with ICI group were better than those in bevacizumab combined with erlotinib group. The median PFS ( P=0.003) and median OS ( P=0.028) in TKI combined with ICI group were better than those in TKI monotherapy group. Conclusions:HLRCC-RCC is rare but has a high degree of malignancy, poor prognosis and familial genetic characteristics. Immunohistochemical staining with strong positive 2-SC and negative FH can provide an important basis for clinical diagnosis. Genetic detection of FH gene germ line mutation can confirm the diagnosis. The preliminary study results confirmed that TKI combined with ICI had a good clinical effect, but it needs to be confirmed by the results of a large sample multi-center randomized controlled clinical study.

OBJECTIVES: This study aimed to analyze the bacteria in dental caries and establish an optimized dental-ca-ries diagnosis model based on 16S ribosomal RNA (rRNA) data of oral flora. METHODS: We searched the public databa-ses of microbiomes including NCBI, MG-RAST, EMBL-EBI, and QIITA and collected data involved in the relevant research on human oral microbiomes worldwide. The samples in the caries dataset (1 703) were compared with healthy ones (20 540) by using the microbial search engine (MSE) to obtain the microbiome novelty score (MNS) and construct a caries diagnosis model based on this index. Nonparametric multivariate ANOVA was used to analyze and compare the impact of different host factors on the oral flora MNS, and the model was optimized by controlling related factors. Finally, the effect of the model was evaluated by receiver operating characteristic (ROC) curve analysis. RESULTS: 1) The oral microbiota distribution obviously differed among people with various oral-health statuses, and the species richness and species diversity index decreased. 2) ROC curve was used to evaluate the caries data set, and the area under ROC curve was AUC=0.67. 3) Among the five hosts' factors including caries status, country, age, decayed missing filled tooth (DMFT) indices, and sampling site displayed the strongest effect on MNS of samples (P=0.001). 4) The AUC of the model was 0.87, 0.74, 0.74, and 0.75 in high caries, medium caries, low caries samples in Chinese children, and mixed dental plaque samples after controlling host factors, respectively. CONCLUSIONS The model based on the analysis of 16S rRNA data of oral flora had good diagnostic efficiency.
Humans ; Child ; Bacteria/genetics* ; Dental Caries/microbiology* ; Dental Caries Susceptibility ; Microbiota/genetics* ; RNA, Ribosomal, 16S

Objective:To investigate the effect of preserved pedicle fascia on the survival of rat posterior intercostal artery perforator pedicle flap.Methods:Fifty SD rats (male and female) were randomly divided into two groups, A and B, with 25 rats in each group. The flap pedicled with the perforating branch of the posterior intercostal artery was designed and cut from the right back of each SD rat, with the size of 10.0 cm×2.5 cm. The perforator branch and the surrounding fascia tissue of 2.5 cm×1.0 cm were retained in group A, while only the perforator branch was retained in group B, the fascia tissue around the perforating branch was removed as much as possible. Finally, the two groups of flaps were sutured in situ. Flap survival of the two groups was observed and the survival rates were calculated. The two groups of flaps were cut and stained with HE, and the angiogenesis of the specimens was observed under a light microscope. Image-Pro Plus 6.0 software was used to calculate the blood vessel density in the sections stained with HE on the 7th day after surgery, vessel diameter and vessel wall thickness were measured and calculated. The expressions of hypoxic-inducible factor-1α (HIF-1α) and cytoglobin (CYGB) in the two groups of flaps were detected by immunohistochemistry. The relative expressions of HIF-1α and CYGB in the two groups were detected by Western blotting, and the differences between the two groups were compared. The measurement data were expressed as Mean±SD. Bilateral t-test was used to compare the data between the two groups, and P<0.05 indicated statistically significant difference. Results:The flaps in group A survived well, while the flaps in group B had distal ischemic necrosis. The survival rate of the flaps in group A (96.19%±0.65%) was higher than that in group B (88.97%±3.24%) at 7 days after operation, and the difference was statistically significant ( P<0.01). HE staining showed neovascularization in both groups. The new blood vessels in group A were higher than those in group B. Blood vessel density, blood vessel diameter and blood vessel wall thickness in group A were (7.77±0.92) vessels/mm 2, (65.23±14.62) μm and (10.49±1.99) μm, respectively, which were higher than those in group B [(5.82±0.85) vessels/mm 2, (40.61±7.78) μm and (6.27±2.63) μm]. The differences were statistically significant( P<0.05). Immunohistochemistry showed that HIF-1α expression in group A was higher than that in group B on day 1 after surgery, and gradually tended to be stable. CYGB expression in group A and Group B peaked on the 7th day after surgery. The results of Western blotting showed that the expression level of HIF-1α in group A was higher than that in group B on the first and third day after surgery, with statistical significance ( P<0.01). HIF-1α in group A reached its peak on the fifth day after surgery, then gradually decreased and tended to be stable. HIF-1α expression in group B was higher than that in group A on the 5th, 7th and 9th day after surgery ( P<0.01), and showed a slow decreasing trend until the 9th day. The expression level of CYGB in group A was higher than that in group B on the third, fifth and ninth day after surgery ( P<0.01), which had statistical significance. The expression of CYGB in group A and Group B reached a peak on day 7 after surgery. Conclusion:The survival rate of the rat posterior intercostal artery perforator flap can be improved by keeping part of fascia in the pedicle, which may be caused by the stable expression of HIF-1α and the upregulated expression of CYGB.

Objective:To investigate the effect of preserved pedicle fascia on the survival of rat posterior intercostal artery perforator pedicle flap.Methods:Fifty SD rats (male and female) were randomly divided into two groups, A and B, with 25 rats in each group. The flap pedicled with the perforating branch of the posterior intercostal artery was designed and cut from the right back of each SD rat, with the size of 10.0 cm×2.5 cm. The perforator branch and the surrounding fascia tissue of 2.5 cm×1.0 cm were retained in group A, while only the perforator branch was retained in group B, the fascia tissue around the perforating branch was removed as much as possible. Finally, the two groups of flaps were sutured in situ. Flap survival of the two groups was observed and the survival rates were calculated. The two groups of flaps were cut and stained with HE, and the angiogenesis of the specimens was observed under a light microscope. Image-Pro Plus 6.0 software was used to calculate the blood vessel density in the sections stained with HE on the 7th day after surgery, vessel diameter and vessel wall thickness were measured and calculated. The expressions of hypoxic-inducible factor-1α (HIF-1α) and cytoglobin (CYGB) in the two groups of flaps were detected by immunohistochemistry. The relative expressions of HIF-1α and CYGB in the two groups were detected by Western blotting, and the differences between the two groups were compared. The measurement data were expressed as Mean±SD. Bilateral t-test was used to compare the data between the two groups, and P<0.05 indicated statistically significant difference. Results:The flaps in group A survived well, while the flaps in group B had distal ischemic necrosis. The survival rate of the flaps in group A (96.19%±0.65%) was higher than that in group B (88.97%±3.24%) at 7 days after operation, and the difference was statistically significant ( P<0.01). HE staining showed neovascularization in both groups. The new blood vessels in group A were higher than those in group B. Blood vessel density, blood vessel diameter and blood vessel wall thickness in group A were (7.77±0.92) vessels/mm 2, (65.23±14.62) μm and (10.49±1.99) μm, respectively, which were higher than those in group B [(5.82±0.85) vessels/mm 2, (40.61±7.78) μm and (6.27±2.63) μm]. The differences were statistically significant( P<0.05). Immunohistochemistry showed that HIF-1α expression in group A was higher than that in group B on day 1 after surgery, and gradually tended to be stable. CYGB expression in group A and Group B peaked on the 7th day after surgery. The results of Western blotting showed that the expression level of HIF-1α in group A was higher than that in group B on the first and third day after surgery, with statistical significance ( P<0.01). HIF-1α in group A reached its peak on the fifth day after surgery, then gradually decreased and tended to be stable. HIF-1α expression in group B was higher than that in group A on the 5th, 7th and 9th day after surgery ( P<0.01), and showed a slow decreasing trend until the 9th day. The expression level of CYGB in group A was higher than that in group B on the third, fifth and ninth day after surgery ( P<0.01), which had statistical significance. The expression of CYGB in group A and Group B reached a peak on day 7 after surgery. Conclusion:The survival rate of the rat posterior intercostal artery perforator flap can be improved by keeping part of fascia in the pedicle, which may be caused by the stable expression of HIF-1α and the upregulated expression of CYGB.

ObjectiveTo analyze the effect of frailty status on the risk of mortality in a community-based population aged 45 years and above in Shanghai with different characteristics， and to provide further basis for population-based interventions for frailty and prevention of adverse outcomes. MethodsData were derived from baseline data from the Shanghai prospective study on AGEing and adult health （2009-2010） and cohort follow-up of causes of death up to October 30， 2021. Frailty index （FI） scores were constructed from 40 variables. Those with frailty index FI≥0.2 were judged to be in a frail state， and a multifactorial Cox regression model was used to calculate the hazard ratio （HR） to evaluate the effect of frailty status on the risk of death in different age groups by gender. Socioeconomic characteristics （age， residence， marital status， education and family economic level， etc.） and health-related behaviors （smoking， alcohol consumption， fruit and vegetable intake， social participation， etc.） were included as control variables. ResultsThe study included 7 978 subjects， 777 （9.7%） of whom were in a frail state. After （11.3±1.8） years of follow-up， 1 043 （13.1%） individuals were dead， including 214 （27.5%） who were frail. The results of the multifactorial Cox regression analysis showed that the effect of frailty on the risk of death in each subgroup was in descending order of men in the middle-aged group （45‒ years） （HR=2.92， 95%CI： 1.38-6.19）， women in the low-aged elderly group （60‒ years） （HR=1.68， 95% CI： 1.08-2.60）， and women in the old-aged elderly group （≥75 years and older） （HR=1.59， 95%CI： 1.22‒2.06）. ConclusionFrailty is associated with the risk of death， and we should focus on the frailty status of men aged 45~59 years and women aged 60 years and above. Early screening and assessment of frailty status and taking appropriate preventive interventions may reduce the occurrence of adverse outcomes and premature death.

Objective:To investigate the present situation of unintended pregnancy within two years postpartum and its influencing factors in China.Methods:Participants who delivered a live birth at 60 hospitals in 15 provinces in the eastern, central and western regions of China during July 2015 to June 2016 were interviewed by using structured questionnaire. Information on occurrence of unintended pregnancy within 2 years after delivery, postpartum contraceptive use, sexual resumption, breastfeeding, and women′s socio-demographic characteristics, and so on, were collected. Life-table analysis, cluster log-rank tests and a 2-level Cox regression model were used for data analysis.Results:A total of 18 045 postpartum women were investigated. The cumulative 1- and 2-year unintended pregnancy rates after delivery were 5.3% (95% CI: 4.5%－6.1%) and 13.1% (95% CI: 11.3%－14.8%), respectively. Cox regression model analysis showed that the risk of unintended pregnancy within 2 years postpartum were increased in younger women, ethnic minorities, women with abortion history, and those who had a vaginal delivery with short lactation time and late postpartum contraceptive initiation (all P<0.01). The risk of postpartum unintended pregnancy was not associated with geographic regions and hospitals where women gave a birth (all P>0.05). Conclusions:In China, the risk of unintended pregnancy within 2 years after delivery is relatively high. Service institutions and service providers should improve the quality of postpartum family planning services, promote the use of high effect contraceptive methods, and educate women to use a method at the time of their sexual resumption or even before.