Objective:To investigate whether long non-coding RNA(lncRNA) AW112010 can improve insulin resistance in aging adipocytes through the miR-204/POU2F2 signaling pathway.Methods:In vivo experiment: C57BL/6 mice were divided into young control group(4 months old) and aging model group(18 months old) based on body weight. The expression levels of AW112010, miR-204-5p, POU2F2, aging related indicators(p16, p21), and insulin signaling pathway genes [insulin receptor(INSR), insulin receptor substrate 1(IRS1), phosphatidylinositol kinase(PI3K), protein kinase B(AKT)] in epididymal adipose tissue were detected using real-time fluorescence quantitative PCR(RT-qPCR) and Western blotting. In vitro experiment: Using adriamycin(ADR) to induce 3T3-L1 aging adipocyte model, β-gal staining was used to observe cellular senescence, and miR-204 inhibitor and miR-204 mimic small interfering RNA were successfully constructed and transfected into 3T3-L1 adipocytes. Results:RT-qPCR and Western blot results showed that compared with the young group, the expression of AW112010 in the adipose tissue of aging mice was increased, while the expression of miR-204-5p was decreased. The expressions of POU2F2, p16, and p21 in the adipose tissue of aging mice were increased, while the expressions of INSR, IRS1, PI3K, GLUT4 mRNA and protein were decreased. The β-gal stainging results showed that the number of 3T3-L1 senescent adipocytes induced by ADR was significantly increased, and the expression levels of AW112010, POU2F2, p16, and p21 in ADR-induced senescent adipocytes were increased compared with the control group, while the expression levels of miR-204-5p, INSR, IRS1, PI3K, GLUT4 were decreased, and remaining glucose in the culture medium was increased. Compared with control, overexpression of miR-204 resulted in decreased expressions of aging indicators p16, p21, and target gene POU2F2 while the expressions of INSR and GLUT4 were increased.Conclusion:Upregulation of lncRNA AW112010 in adipocytes of aging mice may induce insulin resistance by targeting miR-204-5p/POU2F2/IRS1.

OBJECTIVE To investigate the protective effect proanthocyanidin B2(PC-B2) on oxidative damage and apoptosis of mouse astrocytes(AS) induced by hydrogen peroxide(H2O2) and its mechanism. METHODS AS were isolated and cultured from neonatal C57BL/6 mice(1−3 d). The optimal concentration of H2O2 and PC-B2 was divided into four groups: normal group, normal+PC-B2 group(100 μg·mL‒1 PC-B2 treated for 24 h), H2O2 model group(200 μmol·L‒1 H2O2 treated for 24 h), PC-B2 group(200 μmol·L‒1 H2O2 and 100 μg·mL‒1 PC-B2 treated for 24 h). The cell viability of each group was detected by CCK-8 method. Cytotoxicity was detected by LDH method. The antioxidant capacity was detected by ABTS and DPPH. The content of MDA and the activity of SOD, CAT and GSH-Px were detected by ELISA kit. Detection of apoptosis in each group was done by TUNEL staining. The mRNA and protein expression levels of Bax, Bcl-2, Caspase-3, Akt/Stat3, p-Akt, p-Stat3 and Nrf2/HO-1 in AS were detected by RT-PCR and Western blotting, respectively. RESULTS PC-B2 could significantly enhance cell viability and inhibit AS apoptosis. Compared with the H2O2 model group, PC-B2 intervention could significantly reduce the content of LDH and MDA in AS, and increase the activity of SOD, CAT and GSH-Px. PC-B2 intervention could inhibit the mRNA and protein expression of Bax and Caspase-3, and up-regulate the mRNA and protein expression of Akt/Stat3, Bcl-2, Nrf2/HO-1. CONCLUSION PC-B2 can enhance the antioxidant capacity of AS through Akt/Stat3 and Nrf2/HO-1 pathways, therefore reduce H2O2-induced AS oxidative damage and apoptosis.

Objective This study aims to determine whether electroacupuncture regulates the phospholipase C(PLC)/inositol-1,4,5-trisphosphate(PLC/IP3)pathway in platelet-derived growth factor receptor α-positive(PDGFRα+)cells,thereby improving gastrointestinal motility disorders in functional dyspepsia(FD).Methods 40 SD rats were randomly divided into blank group,model group,electroacupuncture group,U73122(PLC inhibitor)group,and U73122+electroacupuncture group,with 8 rats in each group.Except for the blank group,all rats were subjected to multi-factor stress intervention to establish the FD model.After successful modeling,the U73122 group was given intraperitoneal injection of inhibitor,the electroacupuncture group was acupunctured at Zusanli and Taichong points,and the U73122+electroacupuncture group was injected with inhibitor 2 hours before acupunc-ture.Ten days later,gastrointestinal motility was tested;immunoblotting was used to detect the protein expression levels of PDGFRα,PLC,P-PLC,IP3;immunofluorescence was used to detect the average fluorescence density and co-localization expression of PDGFRα and PLC,IP3;electron microscope was used to observe the gap junction(GJ)situation in the gastric antrum area.Results After modeling,the gastrointestinal motility of rats was weakened,the protein expression levels of PDGFRα,PLC,and IP3 were significantly reduced,GJ widened,and cell morphol-ogy changed;compared with the model group,the gastrointestinal motility of rats in the electroacupuncture group,U73122 group,and U73122+electroacupuncture group was significantly improved,the expression levels of PDGFRα,PLC,P-PLC,IP3 increased,GJ was slightly tight,and cell morphology recovered;there was no significant difference in the expression levels of PDGFRα,PLC,P-PLC,IP3 in the gastric antrum of U73122 group and U73122+electroacupuncture group;PDGFRα,PLC,and IP3 had fluorescence co-localization.Conclusion Electroacupuncture can improve gastrointestinal dyskinesia in FD rats by activating the PLC/IP3 pathway in PDGFRα+cells.

Objective To investigate the effect of ethanolic extract of Cichorium glandulosum Boiss.et Huet.on fecal bile acid profiles in high-fat-diet-induced obese mice.Methods Twenty-four 6-week-old C57 BL/6 male mice were divided randomly into normal,model,drug-administration,and metformin groups.Mice in the normal group were fed a regular diet and mice in the other three groups were given high-fat diets.The drug-administration group was gavaged with 10 mL/kg ethanol extract of Cichorium glandulosum Boiss.et Huet.daily,and the metformin group was gavaged with 10 mL/kg metformin daily.After 10 weeks,livers were collected to measure hepatic total triglycerides(TG),total cholesterol(TC),low-density lipoprotein-cholesterol(LDL-C),and high-density lipoprotein(HDL)-C.Feces were collected and analyzed.Results Body weight(P＜0.0001),liver TG(P＜0.05),and TC(P＞0.05)were all significantly higher in model mice compared with normal mice,while LDL-C(P＞0.05)and HDL-C(P＜0.001)were significantly lower,indicating abnormal weight gain and lipid metabolism.Alcoholic extract of Cichorium glandulosum Boiss.et Huet.significantly reduced body weight(P＜0.0001),liver TG(P＜0.0001),serum TG(P＜0.05),TC(P＜0.01),and LDL-C(P＜0.05)in mice.Methodsological validation showed that the current method could accurately quantify 52 bile acids in feces.Analysis of the concentration of each type of bile acid revealed that alcoholic extract of Cichorium glandulosum Boiss.et Huet.significantly increased the secondary/primary bile acid ratio(P＜0.05).Multivariate analysis showed that the bile acid metabolic pattern was significantly altered in all groups.Eight differential bile acids were screened in the drug-administration group relative to the model group using variable importance of projection＞1 and P＜0.05.A search of the Kyoto Encyclopedia of Genes and Genomes database revealed that the differential bile acids were mainly involved in the secondary bile acid biosynthesis pathway.Correlation analysis showed that four differential bile acids,deoxycholic acid(r,=0.6445,P＜0.001),isolithocholic acid(r,=0.5879,P＜0.01),3β-deoxycholic acid(r,=0.6649,P＜0.001),and ω-rhamnoglutaric acid(rs=0.5387,P＜0.01),in feces were strongly positively correlated with body weight.Conclusions Cichorium glandulosum Boiss.et Huet.alcoholic extract may play a role in weight reduction and amelioration of dyslipidemia by modulating secondary bile acid biosynthesis and altering fecal bile acid metabolic profiles.

Multiple sclerosis (MS) is a chronic inflammatory demyelinating disease of the central nervous syestem (CNS) triggered by an autoimmune mechanism, which is the main cause of neurological disability in young people. In MS, a variety of glial cells participate in the pathogenesis and development of the disease, and oligodendrocytes (OL) and myelin cytes are destroyed by autoimmune mediated inflammation. Impaired OL production of oligodendrocyte progenitor cells (OPC) leads to persistent demyelination, myelin fragment accumulation and axonal injury, with clinical manifestation of CNS disability. Microglia (MG), which is involved in the inflammatory response and removal of myelin debris, plays a pivotal role in OPC differentiation and OL maturation, thereby directly or indirectly influencing the process of myelin regeneration. An interactive mechanism exists between these 2 components. The comprehension of the interrelationship between the 2 factors can provide enhanced insights into the pathogenesis of MS and facilitate the development of novel therapeutic strategies, thereby addressing pivotal scientific challenges encountered in clinical management of MS. Therefore, this review summarizes the dynamic changes of microglia in MS and its classic animal models, explores the internal relationship between MG and OL in this process, and focuses on the beneficial effects of targeted regulation of microglia, thereby promoting OPC differentiation and myelin regeneration, in order to provide a new research direction for MS treatment.

Objective:To investigate outcomes and safety of doxycycline-moxifloxacin sequential regimen in the treatment of Mycoplasma genitalium urethritis/cervicitis. Methods:From June 2019 to December 2020, patients with Mycoplasma genitalium urethritis/cervicitis confirmed by nucleic acid amplification testing were successively recruited at Department of Sexually Transmitted Diseases, Hospital of Dermatology, Chinese Academy of Medical Sciences, and received sequential therapy with oral doxycycline for 7 days followed by oral moxifloxacin for 7 days. Clinical and/or etiological assessment was conducted 2 to 3 weeks after the end of treatment. Fisher′s exact test was used to analyze factors influencing the treatment outcome. Results:Totally, 36 eligible subjects were enrolled, including 30 males and 6 females. Among them, 18 (50%) patients completed post-treatment etiological assessment, which showed that 12 achieved microbiological cure, and treatment failures occurred in 6; another 18 patients achieved clinical cure. The overall response rate to doxycycline-moxifloacin sequential therapy was 83.3% (30/36, 95% confidence interval[ CI]: 70.5%, 96.1%) . The treatment outcome showed no significant association with the patients′ age, gender, marital status, number of sexual partners in the past 1 month, history of sexually transmitted diseases, history of antibiotic use in the past 1 month, or co-infections (all P > 0.05) . Conclusion:The efficacy of doxycycline-moxifloacin sequential regimen is limited in the treatment of Mycoplasma genitalium infections in Nanjing area, and clinicians should be alerted to the possibility of treatment failure in clinical practice.

Objective: To isolate a bacteriophage against pan-drug resistant Klebsiella pneumoniae in a burn patient, and to study its biological characteristics, genomic information, and effects on bacterial biofilm. Methods: (1) In 2018, pan-drug resistant Klebsiella pneumoniae UA168 (hereinafter referred to as the host bacteria) solution isolated from the blood of a burn patient in Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine (hereinafter referred to as Ruijin Hospital) was used to isolate and purify the bacteriophage against pan-drug resistant Klebsiella pneumoniae from the sewage of Ruijin Hospital with sewage co-culture method, drip plate method, and double-agar plate method. The bacteriophage was named as phage KP168 and the plaque morphology was observed. (2) The phage KP168 solution was taken for cesium chloride density gradient centrifugation and dialysis, and then the morphology of phage KP168 was observed through transmission electron microscope after phosphotungstic acid negative staining. (3) The phage KP168 solution was taken to determine the lytic ability of the phage KP168 against 20 strains of pan-drug resistant Klebsiella pneumoniae isolated from the burned patients′ blood in Ruijin Hospital by the drip plate method, and then the lysis rate was calculated. (4) The phage KP168 solution at a initial titer of 9.3×10¹¹ plaque-forming unit (PFU)/mL (400 μL per tube) and the host bacteria solution at a concentration of 1×10⁹ colony-forming unit (CFU)/mL (4 mL per tube) were conventionally shaking cultured together for 4 hours at multiplicity of infection (MOI) of 10.000, 1.000, 0.100, 0.010, or 0.001, respectively (1 tube per MOI). The titer of phage KP168 was measured by the double-agar plate method (the measurement method was the same below) to select the optimal MOI. The experiment was repeated three times. (5) The host bacteria solution at a concentration of 1×10⁹ CFU/mL (4 mL per tube) and the phage KP168 solution at an adjusted titer of 5×10⁷ PFU/mL (400 μL per tube) were mixed at the MOI of 0.005. The plaques were counted 0 (immediately), 1, 2, 3, 4, 5, 15, and 30 minutes (1 tube at each time point) after mixing by the double-agar plate method (the counting method was the same below), and the percentage of adsorbed phages was calculated to screen for the optimal adsorption time. The experiment was repeated three times. (6) The host bacteria solution at a concentration of 1×10⁹ CFU/mL (300 μL per tube) and the phage KP168 solution at a titer of 5×10⁸ PFU/mL (60 μL per tube) were mixed at MOI of 0.005 and conventionally shaking cultured after standing for the optimal adsorption time. The phage KP168 titer was measured 0 (immediately), 10, 20, 30, 40, 50, 60, 70, 80, 90, and 100 minutes after culture, and a one-step growth curve was drawn. The experiment was repeated three times. (7) The phage KP168 solution at a titer of 2.5×10¹⁰ PFU/mL was left to stand for 1 hour at 37, 40, 50, 60, or 70 ℃ (3 tubes at each time point, 1 mL per tube) for counting the plaques, and then the thermal stability curve was drawn. SM buffer at a pH values of 5.0, 6.0, 7.0, 7.4, 8.0, 9.0, or 10.0 were added to the phage KP168 solution at a titer of 3.0×10¹⁰ PFU/mL, respectively. The mixed solution was left to stand for 1 hour at 37 ℃ (3 tubes of each pH, each tube containing 100 μL phage KP168 solution and 900 μL SM buffer), and then the plaques were counted, and an acid-base stability curve was drawn. (8) The phage KP168 solution was taken for DNA extraction and sequencing after dialysis as in experiment (2). The whole genome was annotated with Prokka to obtain the coding sequence of phage KP168. Nucleotide′s BLAST function was used to proceed nucleic acid sequence alignment for finding a known phage with the highest similarity to the phage KP168 nucleic acid sequence, and Blastx function was used to translate the coding sequence into protein for its function prediction. The comparison with Antibiotic Resistance Genes Database and Virulence Factors Database was proceeded. (9) In a 96-well plate, at a MOI of 1.000, 0.100, 0.010 or 0.001 (3 wells per MOI), 20 μL phage KP168 solution at a initial titer of 5.8×10¹⁰ PFU/mL was added to 200 μL host bacteria solution at a concentration of 1.5×10⁸ CFU/mL (the same concentration below) for co-cultivation for 48 hours. After 200 μL host bacteria solution was left to stand for 48 hours, 20 μL phage KP168 solution at a titer of 1×10^6, 1×10^7, 1×10^8, 1×10^9, or 1×10¹⁰ PFU/mL (3 wells per titer) was added respectively for action for 4 hours. In both experiments, 200 μL host bacteria solution added with 20 μL SM buffer (3 wells) acted as a negative control, and 220 μL LB culture medium (3 wells) acted as a blank control. Absorbance values were measured by a microplate reader, and inhibition/destruction rates of biofilm were calculated. The experiments were both repeated three times. Results: (1) The plaques of phage KP168 successfully isolated and purified were transparent and round, and its diameter was approximately 1.5 mm. (2) The phage KP168 has a regular polyhedron structure with a diameter of about 50 nm and without a tail. (3) The phage KP168 could lyse 13 of 20 strains of Klebsiella pneumoniae from burned patients, with a lysis rate of 65.0%. (4) When MOI was 1.000, the titer was the highest after co-culturing the phage KP168 with the host bacteria for 4 hours, which was the optimal MOI. (5) After the mixing of the phage KP168 with the host bacteria for 4 minutes, the percentage of the adsorbed phage reached the highest, which was the optimal adsorption time. (6) The one-step growth curve showed that during the lysis of the host bacteria by phage KP168, the incubation period was about 10 minutes, and the lysis period was about 40 minutes. (7) With the condition of 40 ℃ or pH 7.4, the number of plaques and the activity of phage KP168 reached the highest. (8) The genome of phage KP168 was a linear double-stranded DNA with a length of 40 114 bp. There were 48 possible coding sequences. It had the highest similarity to Klebsiella phage_vB_Kp1. The most similar known proteins corresponding to the translated proteins of coding sequences contained 23 hypothetical proteins and 25 proteins with known functions. No resistance genes or virulence factor genes were found. The GeneBank accession number was KT367885. (9) After 48 hours of co-cultivation of the phage KP168 and the host bacteria at each MOI, the inhibition rates of biofilm were similar, with an average of about 45%. After the phage KP168 with a titer of 1×10⁹ PFU/mL acted on the biofilm formed by the host bacteria for 4 h, the destruction rate of biofilm was the highest, reaching an average of 42%. Conclusions In this study, a bacteriophage against pan-drug resistant Klebsiella pneumoniae from a burn patient, phage KP168, is isolated from sewage, which belongs to the tailless phage. It has a wide host spectrum, short adsorption time, and short incubation period, with certain thermal and acid-base stability. Its genomic information is clear, and it does not contain resistance genes or virulence factor genes. It also has an inhibitory effect on the formation of bacterial biofilm and a destructive effect on the formed bacterial biofilm.

Objective: To compare the short-term outcomes of a collagen matrix (CM) and free gingival graft (FGG) in augmenting keratinized mucosa around dental implants. Methods: Nineteen partially edentulous patients who had undergone implant surgery or implant review from June 2017 to June 2018 at Department of Periodontology, Peking University School and Hospital of Stomatology with lack of keratinized mucosa at buccal aspect of implants (<2 mm) were recruited in this study. According to the width of keratinized mucosa (KW) pre-operation, 9 patients including 5 males and 4 females were assigned into control group (KW<0.5 mm) which performed free gingival graft (17 implants) and 10 patients including 3 males and 7 females were assigned into experimental group (KW≥0.5 mm) which used collagen matrix as the grafts (15 implants). The KW at buccal aspect of each implant were measured pre-operation and 2 weeks, 1 month, 2 months, 3 months after surgery respectively. Each of the patients was required to fill out a questionnaire using a visual analogue scale to assess the postoperative morbidity. Results: The KWs around implants were increased significantly during the 3 months follow-up period in both groups (P<0.01). At 3 months after surgery, KW gain in control group was (3.44±1.64) mm, in experimental group was (2.30±0.82) mm, the difference between two groups was statistically significant (P<0.05). Meantime, the total shrinkage of KW in control group [(34±25)%] and experimental group [(51±11)%] also showed a statistically significant difference (P<0.01). However, by using collagen matrix as the grafts, augmented tissues had a much more comparable appearance with adjacent tissues than that in control group. And the patients of experimental group experienced much less postoperative bleeding than those of control group. Conclusions Both collagen matrix and free gingival graft can significantly increase the KW around implants within the 3 months post-surgery follow-up period. There were more KW gain and less shrinkage in group FGG than that in group CM. However, surgery time were reduced and the postoperative bleeding were less in group CM than in group FGG as no harvesting procedure was needed.

Objective:To explore the effects of eight-week group positive psychotherapy intervention on de-pression associated self-efficacy and subjective well-being in female nursing students.Methods:Totally 74 female nursing students with mild to moderate depression [the score of Beck Depression Inventory (BDI-Ⅱ)was 14 to 28]were selected from three nursing colleges of Changsha.They were randomly divided into two groups,40 in the control group and 34 in the intervention group.Samples in the intervention group were given the eight-week group positive psychotherapy,while the control group was given nothing (the control group could receive the intervention when the intervention was proved to be effective or the control group needed treatments).The General Self-efficacy Scale(GSES)and Index of Well-being(IWB)were used to get the outcome of female nursing students'self-effica-cy,subjective well-being at baseline,the time after the intervention,3 month follow-up,6 month follow-up.Results:The interactive effect between group and time of self-efficacy,subjective well-being were statistically significant (Ps<0.05).Separate analyses showed that at baseline,there was no significant difference between two groups in all variables (Ps >0.05).At the end of the intervention,3 month follow-up and 6 month follow-up,the GSES score and IWB score in the intervention group were higher than those in the control group (Ps <0.05).Conclusion:Eight-week group positive psychotherapy could improve self-efficacy and subjective well-being of female nursing students who have mild to moderate depressive symptom.This intervention also has a long-term effect.