BACKGROUND:It has been elucidated that downregulation of nuclear enriched abundant transcript 1(NEAT1)inhibits the progression of keloid fibroblasts,but the exact mechanism is not fully understood. OBJECTIVE:To investigate the influences of long non-coding RNA nuclear enriched abundant transcript 1(lncRNA NEAT1)on the proliferation,apoptosis and migration of keloid fibroblasts by regulating the miR-136-5p/ubiquitin-specific protease 4(USP4)axis. METHODS:Keloid fibroblasts were divided into five groups:si-NC group,control check group,si-NEAT1 group,si-NEAT1+miR-136-5p inhibitor group,and si-NEAT1+inhibitor-NC group.qRT-PCR was performed to measure the expressions of NEAT1 and miR-136-5p;cell counting kit-8 assay and EDU staining were performed to measure cell proliferation;flow cytometry was performed to measure apoptosis;scratch-healing experiment was performed to measure cell migration;western blot assay was performed to measure the protein expressions of USP4,p27,Bax,matrix metalloproteinase-9,α-smooth muscle actin,and type I collagen α1 chain;dual-luciferase assay was performed to examine the relationship of NEAT1 with miR-136-5p as well as the relationship of miR-136-5p with USP4. RESULTS AND CONCLUSION:Compared with the si-NC group,the NEAT1 expression,absorbance value at 450 nm,percentage of EDU positive cells,scratch-healing rate,the protein expressions of USP4,matrix metalloproteinase-9,α-smooth muscle actin,and type I collagen α1 chain decreased in the si-NEAT1 group(P<0.05),while the expression of miR-136-5p,apoptosis rate,and the protein expressions of p27 and Bax increased(P<0.05).miR-136-5p inhibitor reversed the effect of silencing NEAT1 on the biological behavior of keloid fibroblasts.There was a targeted regulatory relationship between NEAT1 and miR-136-5p as well as between miR-136-5p and USP4.To conclude,silencing NEAT1 may inhibit the proliferation and migration of keloid fibroblasts and induce apoptosis by regulating the miR-136-5p/USP4 axis..

As a traditional nutritional and healthy cash crop in China, tea has certain significance in promoting human health and preventing and controlling chronic diseases. Studies have shown that the nutritional health effect of tea is due to its rich functional components, mainly including tea polyphenols, tea pigments, tea polysaccharides, theanine, alkaloids and other bioactive substances. At present, researchers from the academic circles have continuously carried out animal and human experiments on the health effects of various functional components of tea, which has accumulated abundant research data and materials. Based on this, this article reviews the literature on the nutritional and health effects of the main functional components of tea, and adopts the method of evidence-based research to screen and extract relevant data for qualitative and quantitative meta-analysis. Subsequently, the nutritional health effects of the five functional components of tea, namely tea polyphenols, tea pigments, tea polysaccharides, theanine, and alkaloids, are summarized and outlined. Studies have shown that tea polyphenols, tea pigments, tea polysaccharides, theanine and alkaloids have different health effects and are expected to play their unique roles in promoting human health and preventing and controlling diseases.

BACKGROUND:In recent years,the incidence of hyperuricemia caused by purine metabolism disorders has been increasing,which can induce inflammatory responses and lead to renal injury. OBJECTIVE:To explore the role and mechanism of solute carrier family 2 member 12(SLC2A12)in hyperuricemia-related renal injury. METHODS:Renal tubular cells(HK2 cells)were divided into five groups:HK2 group,HK2+uric acid group,HK2+uric acid+NC group,HK2+uric acid+siSLC2A12 group,and HK2+uric acid+siSLC2A12+MK-2206 group.HK2 cells were treated with uric acid and transfected with siRNA SLC2A12,followed by MK-2206 treatment to inhibit AKT expression.Cell proliferation was detected by CCK-8 assay.Apoptosis was detected by TUNEL assay.qRT-PCR and western blot assay were used to detect fibrogenic factors as well as activation of the AKT/FOXO3a pathway.The concentrations of inflammatory cytokines were measured by enzyme-linked immunosorbent assay. RESULTS AND CONCLUSION:(1)Uric acid treatment inhibited cell proliferation and promoted cell apoptosis in the HK2+uric acid group compared with the HK2 group.The proliferative ability of cells in the HK2+uric acid+siSLC2A12 group was further decreased and apoptotic cells were further increased compared with the HK2 group.Compared with the HK2+uric acid+siSLC2A12 group,the HK2+uric acid+siSLC2A12+MK-2206 group showed an increase in cell proliferation and a decrease in apoptotic cells.(2)Compared with the HK2 group,the connective tissue growth factor(CTGF),α-smooth muscle actin(α-SMA)and transforming growth factor beta(TGF-β)expressions increased in the HK2+uric acid group;CTGF,α-SMA and TGF-β expression further increased in the HK2+uric acid+siSLC2A12 group.Compared with the HK2+uric acid+siSLC2A12 group,the CTGF,α-SMA and TGF-β expressions decreased.(3)Compared with the HK2 group,the expression of p-AKT,FOXO3a,and p-FOXO3a elevated in the HK2+uric acid group;the expression of p-AKT further increased,while the expression of FOXO3a and p-FOXO3a decreased in the HK2+uric acid+siSLC2A12 group.Compared with the HK2+uric acid+siSLC2A12 group,p-AKT expression decreased;FOXO3a and p-FOXO3a expression increased in the HK2+uric acid+siSLC2A12+MK-2206 group.(4)Compared with the HK2 group,interleukin-6,interleukin-1 β,and tumor necrosis factor α levels increased in the HK2+uric acid group;interleukin-6,interleukin-1 β,and tumor necrosis factor α levels further increased in the HK2+uric acid+siSLC2A12 group.Compared with the HK2+uric acid+siSLC2A12 group,interleukin-6,interleukin-1 β,and tumor necrosis factor α levels diminished in the HK2+uric acid+siSLC2A12+MK-2206 group.(5)These findings indicate that SLC2A12 may protect against hyperuricemia-induced renal injury by counteracting uric acid-induced tubular fibrosis and inflammation through activation of the FOXO3a pathway.

Propensity score can improve the efficiency and accuracy of clinical trials， and has been widely used in many fields such as comparison of efficacy evaluation， exploration of disease occurrence factors， and monitoring of adverse drug reactions. At present， the propensity scoe has application prospects in traditional Chinese medicine （TCM） monotherapy intervention， complex intervention and integrated Chinese and Western medicine intervention research， and it is a good auxiliary tool for real-world study. By analysing the cha-racteristics of clinical research in TCM， the authors believed that， firstly， the application of propensity score can acce-lerate the progress of clinical efficacy evaluation research in TCM， contribute to the collection of high-level clinical evidence， and promote the development and promotion of TCM therapies； Secondly， under the background of big data resources， propensity score has good application prospects in assisting TCM prescription analysis， drug safety and adverse reaction monitoring， and large-scale cohort studies in TCM； thirdly， in the process of clinical practice， propensity scoring method may face problems such as inappropriate method selection， incorrect judgement or measurement of confounding factors， and unreasonable interpretation or extrapolation of results.

Objective To compare the effects of different doses of vitamin D supplementation on vita-min D deficiency and insufficiency in premature infants.Methods A total of 126 premature infants hospital-ized in the pediatrics department of this hospital from March 25,2021 to December 30,2022 were selected as the study subjects and began to supplement different doses of vitamin D on 3 d after birth under the intestinal tract tolerance.They were divided into 500,900 and 2 100 IU/d groups according to vitamin D supplementary dose.The differences in vitamin D levels at beginning and after supplementation were compared among the groups.Results Among 126 premature infants,there were 52 cases of vitamin D deficiency and 74 cases of vi-tamin D insufficiency.Compared with vitamin D insufficiency,the proportion of vitamin D supplementation in the mothers of the premature infants with vitamin D deficiency was lower(P＜0.05).The serum calcium,phosphorus,alkaline phosphatase and 25-hydroxyvitamin D levels after birth had no statistical difference a-mong 3 groups(P＞0.05).The 25-hydroxyvitamin D level after supplementation had statistical difference a-mong 3 groups(P＜0.05),moreover the 500 IU/d group was lower than the 2 100 IU/d group(P＜0.05).The longest duration of vitamin D supplementation in the 2 100 IU/d group was 90 d,its highest 25-hydroxyvitamin D level after supplementation was 48.3 ng/mL,no vitamin D poisoning condition appeared,there were still 40 cases(85.1％)of vitamin D deficiency and insufficiency after supplementation in the 900 IU/d group,and 17 cases(25.8％)in the 2 100 IU/d group.Conclusion Giving vitamin D 2 100 IU/d in pre-term infants is recommended after birth to prevent the vitamin D deficiency.

Objective To evaluate the diagnostic value of ischemia-modified albumin(IMA)and the crea-tine kinase(CK)index in acute ischemic stroke(AIS).Methods According to the inclusion and exclusion cri-teria,totally 149 newly diagnosed and untreated AIS patients hospitalized in Henan Provincial People's Hospi-tal from October 2021 to October 2022 were selected as the AIS group.Additionally,156 healthy people who underwent the physical examination during the same period were selected as the control group.Activity levels of IMA,CK,creatine kinase-MB(CK-MB),lactate dehydrogenase(LDH)and hydroxybutyrate-dehydrogen-ase(HBDH)were measured using the Abbott C1600 biochemical analyzer,and the CK index(ratio of CK-MB to CK)was calculated.Relative risk factors were analyzed,receiver operating characteristics(ROC)curve was constructed,data were analyzed using SPSS27.0.1,graphs were plotted using GraphPad Prism 9.4.1,and differences in area under the curve(AUC)were compared using MedCalc(version 20.0.22).Results The AIS group exhibited significantly higher levels of IMA,CK-MB,and the CK index,and significantly lower levels of CK compared to the control group(all P＜0.05).Univariate logistic regression analysis revealed that both IMA and the CK index were risk factors for AIS(both P＜0.001).After adjusting for gender and age in a multivariate binary logistic regression analysis,IMA emerged as an independent risk factor for AIS(OR=1.901,95％CI:1.649-2.190,P＜0.001).IMA,CK-MB and CK index in the AIS group were significantly higher than those in the control group,and CK levels were significantly lower than those in the control group,and the differences were statistically significant(P＜0.05).Univariate Logistic regression analysis showed that IMA and CK index were risk factors for AIS(P＜0.001).After adjusting for sex and age in multivariate binary Logistic regression analysis,IMA was an independent risk factor for AIS(OR=1.901,95％CI:1.649-2.190,P＜0.001).The ROC curve demonstrated that AUC,the sensitivity and the specificity of sin-gle detection for IMA were 0.922,81.2％,and 90.4％,respectively.There was no significant difference compared to combined detection using IMA+CK index or IMA+CK index+CK(all P＞0.05).Conclusion IMA is an independent risk factor for AIS,which has strong diagnostic value and is worthy of clinical application.

Objective:To explore the effect of problem-based learning (PBL) combined with case-based learning (CBL) in insulin injection patients with type 2 diabetes mellitus (T2DM) .Methods:From August to October 2022, convenience sampling was used to select 120 patients with T2DM treated with insulin injection from the Endocrinology Department of Yiwu Central Hospital as the study subject. According to the admission time, patients were divided into a control group and an intervention group, with 60 cases each. The 60 patients admitted from August 1st to 31st were in the control group and received routine health education, and 60 patients admitted from October 1st to 31st were included in the intervention group and received PBL combined with CBL. This study analyzed and compared the awareness rate of insulin injection knowledge and assessment scores of insulin injection between two groups of patients, and counted the passing rate and improvement rate of assessment scores after intervention. Before and after three months of intervention, this study evaluated the patient's blood glucose status [fasting blood glucose (FPG), 2-hour postprandial blood glucose (2hPG), and glycated hemoglobin (HbA1c) ], and counted the incidence of insulin injection related complications.Results:After four weeks of intervention, the overall awareness rate of insulin injection knowledge in the intervention group was higher than that in the control group, with a statistically significant difference ( P<0.05). The insulin injection assessment score (90.35±4.68), passing rate of assessment score 91.67%, and improvement rate of assessment score (35.48±6.84) % in the intervention group were higher than those [ (84.69±6.72), 75.00%, and (26.28±5.46) %] in the control group, and the differences were statistical ( P<0.05). After three months of intervention, the FPG, 2hPG, and HbA1c of the intervention group patients were lower than those of the control group, with statistically significant differences ( P<0.05). Within three months after intervention, the total incidence of complications in the intervention group was lower than that in the control group, and the difference was statistically significant ( P<0.05) . Conclusions:PBL combined with CBL for insulin injection patients can help to increase their knowledge and operational level of insulin injection, improve blood glucose control, and reduce insulin injection related complications.

Humans ; COVID-19/genetics* ; SARS-CoV-2/genetics* ; Clustered Regularly Interspaced Short Palindromic Repeats/genetics* ; CRISPR-Cas Systems/genetics* ; RNA, Viral/genetics*

Tyrosol is a natural polyphenolic product that is widely used in chemical, pharmaceutical and food industries. Currently, the de novo synthesis of tyrosol by Escherichia coli suffers from issues such as low cell density and poor yield. Therefore, the phenylpyruvate decarboxylase mutant ARO10^F138L/D218G obtained in our previous study was fused with an alcohol dehydrogenase from different microorganisms for fusion expression, and the optimal ARO^{10F138L/D218G}-L-YahK produced 1.09 g/L tyrosol in shake flasks. In order to further improve tyrosol production, feaB, a key gene in the competing pathway of 4-hydroxyphenylacetic acid, was knocked out, and the resulted strain produced 1.26 g/L tyrosol with an increase of 21.15% compared to that of the control. To overcome the low cell density in tyrosol fermentation, the quorum-sensing circuit was used to dynamically regulate the tyrosol synthesis pathway, so as to alleviate the toxic effect of tyrosol on chassis cells and relieve the growth inhibition. Using this strategy, the yield of tyrosol was increased to 1.74 g/L, a 33.82% increase. In a 2 L fermenter, the production of tyrosol in the engineered strain TRFQ5 dynamically regulated by quorum-sensing reached 4.22 g/L with an OD₆₀₀ of 42.88. Compared with those in the engineered strain TRF5 statically regulated by induced expression, the yield was increased by 38.58% and the OD₆₀₀ was enhanced by 43.62%. The combination of blocking the competing pathway using gene knockout technology, and reducing the inhibitory effect of tyrosol toxicity on chassis cells through quorum-sensing dynamic regulation increased the production of tyrosol. This study may facilitate the biosynthesis of other chemicals with high toxicity.
Escherichia coli/genetics* ; Biological Products ; Bioreactors ; Fermentation

ObjectiveTo explore the anti-tumor effect and mechanism of Shenqi Yiliu prescription in the intervention of pyroptosis. MethodTen male BALB/c mice were randomly selected and assigned to the blank group. The remaining 40 mice underwent the induction of the liver cancer xenograft model. After 5 days of modeling， 40 surviving mice were randomly divided into model group， cisplatin group ［2.5×10^-3 g·kg^-1·（3 d）^-1］， Shenqi Yiliu prescription group （27 g·kg^-1·d^-1）， and a combination group （Shenqi Yiliu prescription group + cisplatin）. The mice in the blank group and the model group were treated with an equal volume of normal saline for 10 days. The general conditions of mice in each group were observed. After the intervention， the tumor weight of the mice was weighed and the tumor inhibition rate was calculated. Hematoxylin-eosin （HE） staining was used to observe the pathological changes in tumor tissues. The levels of mouse liver function indicators， including alanine aminotransferase （ALT） and aspartate aminotransferase （AST） were detected. The TdT-mediated dUTP-biotin nick end labeling （TUNEL） assay was used to detect DNA damage in mouse tumor tissue cells. Immunohistochemistry （IHC）， immunofluorescence （IF）， and Western blot were used to detect the protein expression levels of NOD-like receptor protein 3 （NLRP3）， cysteinyl aspartate-specific protease-1 （Caspase-1）， and gasdermin D （GSDMD） in tumor tissues. The levels of interleukin-1β （IL-1β） and interleukin-18 （IL-18） in tumor tissues were detected by enzyme-linked immunosorbent assay （ELISA）. ResultCompared with the mice in the blank group， those in the model group were in a poor mental state， sleepy， and lazy， and their fur color was dull， with increased levels of serum ALT and AST in liver function tests （P<0.01）. Compared with the model group， the groups with drug intervention showed improved mental state， inhibited tumor growth to varying degrees， and decreased tumor weight， and the tumor inhibition rate in the combination group was the highest （P<0.01）. HE staining showed that the pathological and morphological lesions of the tumor tissues in the model group were significant， while those in all groups with drug intervention were improved to a certain extent. The karyolysis and nuclear rupture in the Shenqi Yiliu prescription group and the combination group were more significant. In the liver function test， the serum ALT and AST levels of mice in the Shenqi Yiliu prescription group and the combination group decreased （P<0.01）， and the inflammatory factors IL-1β and IL-18 in each group with drug intervention decreased （P<0.05， P<0.01）. Among them， the declining trend of IL-1β and IL-18 in the Shenqi Yiliu prescription group was the most significant （P<0.01）. TUNEL staining showed that the positive TUNEL staining in each group with drug intervention decreased after intervention （P<0.05， P<0.01）， especially the cisplatin group and Shenqi Yiliu prescription group （P<0.01）. Western blot， IHC， and IF found that the protein expression levels of NLRP3， Caspase-1， and GSDMD in each group with drug intervention decreased （P<0.05， P<0.01）. Compared with the mice in the cisplatin group， those in the Shenqi Yiliu prescription group and the combination group had better mental state and regular tumor morphology， and the tumor weight of the mice in the combination group decreased （P<0.05）. The levels of ALT and AST in the Shenqi Yiliu prescription group decreased （P<0.05）， and the levels of IL-1β and IL-18 in the Shenqi Yiliu prescription group and the combination group decreased （P<0.05， P<0.01）， especially in the combination group （P<0.01）. The results of IHC showed that the expression of GSDMD protein in the tumor tissues of mice in the combination group was reduced （P<0.01）. IF detection showed that the expression of NLRP3 in the tumor tissues of the Shenqi Yiliu prescription group was reduced （P<0.01）. The results of Western blot showed that the expression level of NLRP3 protein in the Shenqi Yiliu prescription group and the combination group decreased （P<0.01）， and the expression level of Caspase-1 protein in the combination group decreased （P<0.01）. The decrease in GSDMD protein expression was not significant， and the difference was not statistically significant. ConclusionShenqi Yiliu prescription combined with cisplatin has an obvious anti-tumor effect， which may be achieved by down-regulating the NLRP3/Caspase-1/GSDMD inflammatory pyroptosis pathway to inhibit cell pyroptosis， and relieve the inflammatory response in mice with liver cancer.