Objective:To establish a rapid,quantitative and visualized method for the rapid detection of Streptococcus mutans(S.mutans)based on loop-mediated isothermal amplification(LAMP).Methods:Nucleic acid of S.mutans was extracted for LAMP reaction,the detection experimental conditions were optimized.Finally the positive results of the experiments were quantita-tively analyzed by using the mobile-based detection equipment.Results:The system detected the samples within 23 min and showed good specificity and sensitivity with an accuracy of 1.625×10-3 ng/μL.Positive results were captured,identified and quantified u-sing a mobile application,and the quantified results showed a good linear relationship between the absolute G value and the DNA concentration,with an equation of y=3.2x+57.133(R2=0.988 2).Conclusion:Quantitative detection of S.mutans by LAMP has the characteristics of high sensitivity,high specificity,result visualization,convenience and time-saving,and can quantify bacterial DNA.It is a new method for the early detection of caries susceptibility,and can be widely used in clinic.

Objective:To establish a rapid,quantitative and visualized method for the rapid detection of Streptococcus mutans(S.mutans)based on loop-mediated isothermal amplification(LAMP).Methods:Nucleic acid of S.mutans was extracted for LAMP reaction,the detection experimental conditions were optimized.Finally the positive results of the experiments were quantita-tively analyzed by using the mobile-based detection equipment.Results:The system detected the samples within 23 min and showed good specificity and sensitivity with an accuracy of 1.625×10-3 ng/μL.Positive results were captured,identified and quantified u-sing a mobile application,and the quantified results showed a good linear relationship between the absolute G value and the DNA concentration,with an equation of y=3.2x+57.133(R2=0.988 2).Conclusion:Quantitative detection of S.mutans by LAMP has the characteristics of high sensitivity,high specificity,result visualization,convenience and time-saving,and can quantify bacterial DNA.It is a new method for the early detection of caries susceptibility,and can be widely used in clinic.

Objective:To investigate the mechanism of Sulfo-N-succinimidyloleate (SSO) regulating lipid metabolism disorder induced by silicon dioxide (SiO 2) . Methods:In March 2023, Rat alveolar macrophages NR8383 were cultured in vitro and randomly divided into control group (C), SSO exposure group (SSO), SiO 2 exposure group (SiO 2) and SiO 2+SSO exposure group (SiO 2+SSO). NR8383 cells were exposure separately or jointly by SSO and SiO 2 for 36 h to construct cell models. Immunofluorescence and BODIPY 493/ 503 staining were used to detect cluster of differentiation (CD36) and intracellular lipid levels, the protein expression levels of CD36, liver X receptors (LXR), P-mammalian target of rapamycin (P-mTOR) and cholinephosphotransferase 1 (CHPT1) were detected by Western blot, respectively, and lipid metabolomics was used to screen for different lipid metabolites and enrichment pathways. Single-factor ANOVA was used for multi-group comparison, and LSD test was used for pair-to-group comparison. Results:SiO 2 caused the expression of CD36 and P-mTOR to increase ( P=0.012, 0.020), the expression of LXR to decrease ( P=0.005), and the intracellular lipid level to increase. After SSO treatment, CD36 expression decreased ( P=0.023) and LXR expression increased ( P=0.000) in SiO 2+SSO exposure group compared with SiO 2 exposure group. Metabolomics identified 87 different metabolites in the C group and SiO 2 exposure group, 19 different metabolites in the SiO 2 exposure group and SiO 2+SSO group, and 5 overlaps of different metabolites in the two comparison groups, they are PS (22∶1/14∶0), DG (O-16∶0/18∶0/0∶0), PGP (i-13∶0/i-20∶0), PC (18∶3/16∶0), and Sphinganine. In addition, the differential metabolites of the two comparison groups were mainly concentrated in the glycerophospholipid metabolism and sphingolipid metabolism pathways. The differential gene CHPT1 in glycerophospholipid metabolic pathway was verified, and the expression of CHPT1 decreased after SiO 2 exposure. Conclusion:SSO may improve SiO 2-induced lipid metabolism disorders by regulating PS (22∶1/14∶0), DG (O-16∶0/18∶0/0∶0), PGP (i-13∶0/i-20∶0), PC (18∶3/16∶0), SPA, glycerophospholipid metabolism and sphingolipid metabolism pathways.

Objective:To investigate the mechanism of Sulfo-N-succinimidyloleate (SSO) regulating lipid metabolism disorder induced by silicon dioxide (SiO 2) . Methods:In March 2023, Rat alveolar macrophages NR8383 were cultured in vitro and randomly divided into control group (C), SSO exposure group (SSO), SiO 2 exposure group (SiO 2) and SiO 2+SSO exposure group (SiO 2+SSO). NR8383 cells were exposure separately or jointly by SSO and SiO 2 for 36 h to construct cell models. Immunofluorescence and BODIPY 493/ 503 staining were used to detect cluster of differentiation (CD36) and intracellular lipid levels, the protein expression levels of CD36, liver X receptors (LXR), P-mammalian target of rapamycin (P-mTOR) and cholinephosphotransferase 1 (CHPT1) were detected by Western blot, respectively, and lipid metabolomics was used to screen for different lipid metabolites and enrichment pathways. Single-factor ANOVA was used for multi-group comparison, and LSD test was used for pair-to-group comparison. Results:SiO 2 caused the expression of CD36 and P-mTOR to increase ( P=0.012, 0.020), the expression of LXR to decrease ( P=0.005), and the intracellular lipid level to increase. After SSO treatment, CD36 expression decreased ( P=0.023) and LXR expression increased ( P=0.000) in SiO 2+SSO exposure group compared with SiO 2 exposure group. Metabolomics identified 87 different metabolites in the C group and SiO 2 exposure group, 19 different metabolites in the SiO 2 exposure group and SiO 2+SSO group, and 5 overlaps of different metabolites in the two comparison groups, they are PS (22∶1/14∶0), DG (O-16∶0/18∶0/0∶0), PGP (i-13∶0/i-20∶0), PC (18∶3/16∶0), and Sphinganine. In addition, the differential metabolites of the two comparison groups were mainly concentrated in the glycerophospholipid metabolism and sphingolipid metabolism pathways. The differential gene CHPT1 in glycerophospholipid metabolic pathway was verified, and the expression of CHPT1 decreased after SiO 2 exposure. Conclusion:SSO may improve SiO 2-induced lipid metabolism disorders by regulating PS (22∶1/14∶0), DG (O-16∶0/18∶0/0∶0), PGP (i-13∶0/i-20∶0), PC (18∶3/16∶0), SPA, glycerophospholipid metabolism and sphingolipid metabolism pathways.

AIM: To observe the analgesic effect of esketamine in patients with thoracoscopic lobectomy. METHODS: Sixty patients scheduled with thoracoscopic lobectomy were randomly divided into group esketamine (ESK, n =30) and group saline (SAL, n = 30). Esketamine in ESK group was given 0.2 mg/kg at induction and 0.12 mg • kg

Background Lipid metabolism imbalance is tightly linked to the development and progression of multiple diseases. The phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin (PI3K/AKT/mTOR) signaling pathway is important for the regulation of lipid metabolism. However, whether silicosis is associated with lipid metabolic abnormalities has yet to be explored. Objective To observe the changes of lipid deposition, cholesterol, and phosphorylated proteins of PI3K/AKT/mTOR pathway in silicon dioxide (SiO₂)-induced MLE-12 cells and to explore potential mechanism of lipid composition regulated though the pathway. Methods (1) MLE-12 cells were stimulated with 50 mg·L⁻¹ SiO₂ suspension, and divided into fourgroups: a control group and three SiO₂ groups (12, 24, and 48 h of stimulation). (2) Cellproliferation was detected to determine an optimal dose of LY294002, an inhibitor of PI3K protein. LY294002 at 5 μmol·L⁻¹ was used for further study, in which MLE-12 cells cultured for 48 h were divided into four groups: a control group; a 50 mg·L⁻¹ SiO₂ suspension stimulation group; a 50 mg·L⁻¹ SiO₂ suspension and 5 μmol·L⁻¹ LY294002 treatment group; a 5 μmol·L⁻¹ LY294002 treatment group. Total cholesterol (TC), free cholesterol (FC), cholesterol ester (CE; total cholesterol minus free cholesterol), and triglycerides (TG) were measured with enzyme assay kits. Lipid deposition was observed using Oil Red O staining. The expressions of p-PI3K, p-AKT, and p-mTOR proteins were detected by Western blotting. Results (1) The contents of TC, FC, and CE in the 50 mg·L⁻¹ SiO₂-induced MLE-12 cells were increased compared to those of the control group in a time-dependent manner by trend analysis, and the increment at 24 and 48 h were significant. By 48 h, the contents of cholesterol indicators were all elevated: TC from (2.242±0.181) mg·g⁻¹ to (5.148±0.544) mg·g⁻¹, FC from (1.923±0.158) mg·g⁻¹ to (4.168±0.433) mg·g⁻¹, and CE from (0.318±0.067) mg·g⁻¹ to (0.978±0.134) mg·g⁻¹, compared with the control group (P<0.01). The changes of TG were not significant (P>0.05). The SiO₂ suspension induced orange-red particle deposition in the MLE-12 cells, especially at 48 h (P<0.01). The protein expression levels of p-PI3K, p-AKT, and p-mTOR in SiO₂-stimulated MLE-12 cells were higher than those of the control groups with the prolongation of stimulation time, which peaked at 48 h (P<0.01). (2) The contents of TC, FC, and CE in MLE-12 cells of the SiO₂ + LY294002 group were decreased, comparing to those of the SiO₂ stimulation only group (P<0.01), companied with less orange-red lipid deposition, and suppressed protein expression levels of p-PI3K, p-AKT, and p-mTOR (P<0.01). Conclusion SiO₂ could induce increases of cholesterol and lipid deposition through activation of PI3K/AKT/mTOR signaling pathway in MLE-12 cells.

Objective To construct a human renal epithelial cell line HEK293T by CRISPR-Cas9-based site-directed knock-in of vascular endothelial growth factor 165 (VEGF165) gene, and avoid the off-target effect caused by lentivirus infection. Methods The VEGF165 expression vector with homologous arm (pUCm-T-VEGF165 plasmid) and the sgRNA expression vector [pSpCas9(BB)-2A-Puro-sgRNA plasmid] were designed and constructed based on the DNA sequence of the EZH2 gene, and then co-transfected into HEK293T cells. The expression of VEGF165 mRNA was detected by qPCR and the expressions of VEGF165 proteins were detected by Western Blot. Results The qPCR and Western Blot results showed that, comparing with the control, the pUCm-T-VEGF165 plasmid and pSpCas9(BB)-2A-Puro-sgRNA plasmid, the expression of the co-transfection plasmid were significantly increased, i.e. 3.42±0.30 vs. 1.02±0.21, 1.13±0.16 and 0.98±0.18 for the VEGF165 mRNA level (all P<0.01), and 1.13±0.16 vs. 1.02±0.06, 0.88±0.03 and 0.80±0.05 for the VEGF165 protein level (all P<0.01), respectively. Besides, the expression of EZH2 was significantly down-regulated, i.e. 0.14±0.06 vs. 1.08±0.11, 1.02±0.12 and 1.13±0.16 for the EZH2 mRNA level (all P<0.01), and 0.23±0.03 vs. 1.05±0.13, 0.91±0.04 and 0.81±0.06 for the EZH2 protein level (all P<0.01), respectively. This result showed that the VEGF165 was successfully inserted into the EZH2 genome, interfering the EZH2 expression. Conclusions VEGF165 gene can be successfully knocked into HEK293T cells by CRISPR/Cas9 system.

Objective To compare the clinical features and prognosis between dermatomyositis-associated interstitial lung disease (DM-ILD) and idiopathic pulmonary fibrosis (IPF).Methods Patients with interstitial lung disease with dermatomyositis (DM-ILD) or idiopathic pneumonia fibrosis (IPF) from January 2003 to March 2014 in the third affiliated hospital of Sun Yat-sen University were included.Results Among the 64 patients enrolled,44 were DM-ILD and 20 were IPF.IPF was more common in the elderly (P =0.000),men (P =0.004) and smokers (P =0.000),and its high-resolution computed tomography (CT) mostly showed grid shadow (P=0.014) and honeycomb shadow (P=0.000).DM-ILD usually had cough symptoms (P =0.025).High-resolution CT showed patchy (P =0.048) and banded (P =0.000).Glucocorticoid (P =0.000) and immunosuppressive agents (P =0.000) were commonly used in the treatment of DM-ILD.However,there was no significant difference in 90d mortality between the two groups (P ＞ 0.05).Conclusions IPF is more common in the elderly,men and smokers,and its high-resolution CT mostly shows grid shadow and honeycomb shadow,distribution is diffuse.DM-ILD often has cough symptoms,and its high resolution CT is mostly plaques and streaky shadows.Glucocorticoids and immunesuppressants are commonly used in DM-ILD,but there is no significant difference in 90-day mortality between them.

Objective To analyze the rules of acupoint selection in treating knee osteoarthritis with acupuncture and moxibustion therapy based on data mining; To provide references for clinical treatment.MethodsArticles about treating knee osteoarthritis with acupuncture and moxibustion were retrived from China National Knowledge Infrastructure (CNKI), Wanfang academic periodical full-text database, Chinese Journal Full-text Database (VIP) and SinoMed in China from Jan. 1st 2006 to Jan. 1st 2016. The database of acupuncture and moxibustion prescription was set up. The Apriori association rules, system cluster analysis and factor analysis were adopted in order to analyze the rules of acupoint selection for knee osteoarthritis.ResultsTotally 190 articles were included, covering 228 acupuncture- moxibustion prescriptions, 65 acupoints and the total frequency for 1736 times. The usual acupoints in the lower limbs were Dubi (ST35), Neixiyan (EX-LE 4), and Yanglingquan (GB34). The main maridians were Stomach Meridian of Foot-Yangming, Spleen Meridian of Foot-Taiyin, and Gallbladder Meridian of Foot-Shaoyang, accounting to for 1626 times (93.66%). Cluster analysis achieved other four groups of acupoint selection based on syndrome differentiation.ConclusionLocal acupoints and dialectical point selection were important principle in treating knee osteoarthritis combined with acupoint selection based on syndrome differentiation.

Objective To detect the expression of NRP‐1 in gastric cancer tissue ,to analyze its relationship with clinicopath‐ological features ,and to explore its value in judging the prognosis of gastric cancer .Methods The clinical pathologic data and prog‐nosis situation in 168 cases of gastric cancer were retrospectively analyzed .The expression of NRP‐1 in gastric cancer tissue and normal tissue was detected by the immunohistochemical method .The relationship between the expression of NRP‐1 with the clinico‐pathological features and prognosis was investigated .Whether NRP‐1 serving as a reference indicator for judging the prognosis of gastric cancer was evaluated .Results (1) In 168 cases ,the NRP‐1 expression in gastric cancer tissue was higher than that in nor‐mal tissue (66 .7% vs .8 .33% ,P<0 .05) .(2) The NRP‐1 expression was related with the tumor size ,differentiation degree ,infil‐trative depth ,lymph node metastasis and TNM stage(P<0 .05) .(3)The median survival time in the patients with high NRP‐1 ex‐pression was shorter than that in the patients with low NRP‐1 expression (P<0 .05) .(4) The multiple factor analysis by COX pro‐portional hazard model showed that the NRP‐1 expression was an independent risk factor for the prognosis of gastric cancer(P<0 .05) .Conclusion (1)NRP‐1 plays an important role in the incidence and development process of gastric cancer and its expression is closely related with the malignant biological behavior of gastric cancer .(2)The high NRP‐1 expression prompts poor prognosis .(3) NRP‐1 may be expected to be regarded as one of the indexes for judging the biologic behaviors and prognosis of gastric cancer .