BACKGROUND:The diabetic microenvironment can cause excessive M1 polarization of macrophages,and this hyperglycemic inflammatory state can inhibit osteogenic differentiation of bone marrow mesenchymal stem cells,thus affecting the healing of diabetic bone defects.Studies have indicated that mogroside V possesses anti-inflammatory,antioxidant,and hypoglycemic properties.However,its potential to modulate M1 polarization of macrophages and osteogenic differentiation of bone marrow mesenchymal stem cells under high glucose and inflammatory condition remains unclear. OBJECTIVE:To explore the effect of mogroside V on regulating M1 macrophage polarization and its effect on osteogenic differentiation of bone marrow mesenchymal stem cells under high glucose and inflammatory condition. METHODS:Murine diabetic models were established using C57BL/6 mice.Bone marrow-derived macrophages were isolated from tibia and fibula of normal and diabetic mice,and cultured in low-glucose and high-glucose media.Then M1 polarization of bone marrow-derived macrophages was induced using lipopolysaccharide and interferon-γ.Bone marrow-derived macrophages were treated with 160,320,and 640 μmol/L mogroside V.Flow cytometry was employed to determine the proportion of F4/80+CD86+cells.qRT-PCR was utilized to assess mRNA expression levels of inducible nitric oxide synthase,interleukin 1β,and interleukin 6.ELISA was employed to evaluate tumor necrosis factor-α secretion in bone marrow-derived macrophage supernatants.Bone marrow mesenchymal stem cells were isolated from tibia and fibula of C57BL/6 suckling mice,and induced osteogenic differentiation using low-or high-glucose osteogenic induction medium.Bone marrow mesenchymal stem cells were treated with M1 macrophage-conditioned mediums with or without 320 μmol/L mogroside V in osteogenic differentiation process.qRT-PCR was employed to assess the mRNA expression of alkaline phosphatase,Runt-related factor 2,osteocalcin,and osteopontin on day 14 after osteogenic induction.Alizarin red staining and quantitative analysis were conducted to evaluate calcium deposition on day 21 after osteogenic induction. RESULTS AND CONCLUSION:(1)Flow cytometry results showed that with the treatment of 320 and 640 μmol/L mogroside V,the proportion of F4/80+CD86+bone marrow-derived macrophages was significantly lower than that in the high-glucose control group(P<0.05).(2)qRT-PCR results showed that with the treatment of 160,320,and 640 μmol/L mogroside V,the mRNA expression levels of inducible nitric oxide synthase and interleukin 6 were significantly lower than that in the high-glucose control group(P<0.05).With the treatment of 320 and 640 μmol/L mogroside V,the mRNA expression level of interleukin 1β was significantly lower than that in the high-glucose control group(P<0.05).(3)ELISA results exhibited that with the treatment of 160,320,and 640 μmol/L mogroside V,the tumor necrosis factor-α secretion level was significantly lower than that in the high-glucose control group(P<0.05).(4)With the treatment of 320 μmol/L mogroside V,calcium salt deposition was increased in bone marrow mesenchymal stem cells under high glucose and inflammatory conditions(P<0.05),and the mRNA relative expression levels of alkaline phosphatase,Runt-related factor 2,and osteopontin were increased(P<0.05).These findings indicate that mogroside V can promote osteogenic differentiation of bone marrow mesenchymal stem cells by inhibiting the M1 polarization of bone marrow-derived macrophages under high glucose and inflammatory conditions and reducing the generation of inflammatory factors.

OBJECTIVES: To explore the mechanism of cinnamic acid (CA) for improving doxorubicin-induced myocardial injury (DIC) in mice. METHODS: Network pharmacology analysis was used to obtain the key targets of CA and DIC. Male C57BL/6J mice were randomized into Sham, DOX, CA (25, 50 and 100 mg/kg)+DOX, and CA+Ferrostatin-1+DOX groups, and their myocardial function and pathology were examined by echocardiography and HE staining. Serum levels of CK-MB, LDH, MDA, IL-6, TNF‑α and myocardial ROS level were detected, and the expression levels of TLR4 and ferroptosis pathway proteins in myocardial tissue were detected by Western blotting. Cultured murine cardiomyocytes (HL-1 cells) with or without transfection with a small interfering RNA targeting TLR4 (si-TLR4) were treated with DOX or Erastin, and the cellular ROS content was measured by DCFH-DA staining; the expression level of GPX4 was detected using immunofluorescence staining. RESULTS: Network pharmacology analysis suggested that CA may improve DIC through TLR4 signaling. DOX treatment caused obvious myocardial injury in mice, which showed significantly increased serum levels of CK-MB, LDH, MDA, IL-6, TNF-α and myocardial ROS level with decreased myocardial levels of SLC7A11 and GPX4 proteins and increased levels of TLR4 and PTGS2 proteins. All these changes in the mouse models were significantly alleviated by treatment with CA, and the mice receiving CA or ferrostatin-1 treatment exhibited increased myocardial expressions of SLC7A11 and GPX4 proteins and lowered expressions of TLR4 and PTGS2 proteins. In cultured HL-1 cells, treatment with DOX and Erastin both obviously increased intracellular ROS level and decreased cellular GPX4 expression level, and these changes were strongly attenuated by TLR4 interference. CONCLUSIONS CA, as a potent herbal monomer, can effectively alleviate DIC in mice by inhibiting TLR4-mediated ferroptosis.
Animals ; Ferroptosis/drug effects* ; Toll-Like Receptor 4/metabolism* ; Myocytes, Cardiac/metabolism* ; Mice, Inbred C57BL ; Mice ; Male ; Doxorubicin/adverse effects* ; Cinnamates/pharmacology* ; Signal Transduction ; Reactive Oxygen Species/metabolism*

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Objective To explore the value of contrast-enhanced ultrasound(CEUS)in the assessment of diabetic microangiopademia through evaluating microcirculation perfusion of triceps surae muscle by CEUS.Method Totally 51 patients with type 2 diabetes mellitus(T2DM)admitted in our hospital between August 2020 and January 2023 were collected,including 15 pure T2DM patients and 36 T2DM patients complicated with microcirculatory disturbance(T2DM+CM).Each patient's hemoglobin A1c(HbA1c)and homeostatic model assessment for insulin resistance(HOMA-IR)were recorded.After getting enhanced intensity(PI-BI)and regional peak time(TTP-AT)by CEUS,comparative analysis between groups was conducted.Results The levels of HbA1c and HOMA-IR in T2DM+CM group were higher than those in pure T2DM group(P<0.05).TTP-AT in T2DM+CM group were longer than that in pure T2DM group of all muscles musculi gastrocnemii(MG),laterale musculi gastrocnemi(LG),soleus(SOL)and triceps surae muscle's junction region)(P<0.05).The TTP-AT of SOL was longest in both groups,followed by LG,and MG(P<0.05).The PI-BI had no significant difference among MG,LG and SOL in pure T2DM group.The PI-BI of MG was higher than that of SOL in the T2DM+CM group(P<0.05).TTP-AT of triceps surae muscle's junction region had significant positive association with both HbA1c and HOMA-IR(P<0.05).Conclusion The TTP-AT of triceps surae muscle measured by CEUS is a new indicator for evaluating microangiopathopathy in T2DM patients.

Objective To analyze the influencing factors of postoperative urinary tract infection in patients undergoing transurethral resection of the prostate with plasmakinetic energy(PKRP)and establish a risk prediction nomogram model.Methods The data of PKRP patients in Department of Urology,the Second Affiliated Hospital of Nanchang University from December 2020 to September 2021 were selected as the modeling set,and the high-risk factors were screened by univariate analysis and Logistic regression analysis.The risk prediction nomogram model was constructed and verified internally and externally.Results The incidence of urinary tract infection after PKRP surgery was 15.38%.Multivariate analysis showed that age,other location infection,diabetes,preoperative catheterization,urethral injury,indwelling catheter material,hair coloring catheter replacement times and number of indwelling catheterization were risk factors for urinary tract infection(P<0.05).Internal verification(area under the curve was 0.875)and external verification(area under the curve was 0.869)show that the risk prediction nomogram model has good discrimination and accuracy.Conclusion The influencing factors of urinary tract infection after PKRP are complex.The risk prediction nomogram model has good prediction performance,which can provide a basis for the prevention and treatment of urinary tract infection after PKRP.

Objective:To analyze the medication law of Professor Li Fazhi in the prescriptions for the treatment of cough; To explore his academic thoughts.Methods:Medical cases of Professor Li about the treatment for cough from January 1, 2015 to October 31, 2022 were collected. Excel2016 and R language 4.2.1 were used to conduct multidimensional analysis on property and taste, and meridians of drugs. High-frequency efficacy classification was explored through factor analysis, clustering analysis was conducted to distinguish drug groups, and time-lapse analysis on proportion and meridian was conducted on high-frequency drugs.Results:4 746 prescriptions involved 270 kinds of Chinese materia medica, with a total frequency of 57 700 times. The most common property and taste was warm, followed by cold. Warm medicines were mainly pungent warm and cold warm, and cold medicines were mainly pungent cold, sweet cold and bitter cold, and the main meridians were lung, spleen, stomach, and liver meridians. The top 35 kinds of Chinese materia medica with frequency could be clustered into 9 groups. Group 1: Perillae Fructus, Armeniacae Semen Amarum and Ephedrae Herba; group 2: Magnoliae Flosmagnoliae Flos, Cicadae Periostracum and Glycyrrhizae Radix et Rhizoma; group 3: Peucedani Radix, Platycodonis Radix, Mori Cortex, Schizonepetae Herba, Saposhnikoviae Radix and Farfarae Flos; group 4: Angelicae Dahuricae Radix, Notopterygii Rhizoma et Radix, Citri Reticulatae Pericarpium, Astragali Radix, Cimicifugae Rhizoma and Coptidis Rhizoma; group 5: Coicis Semen, Phragmitis Rhizoma, Persicae Semen and Benincasae Semen; group 6: Perillae Folium; group 7: Bupleuri Radix, Mume Fructus, Aurantii Fructus Immaturus, Scutellariae Radix and Paeoniae Radix Alba; group 8: Asari Radix et Rhizoma, Cinnamomi Ramulus, Zingiberis Rhizoma, Schisandrae Chinensis Fructus and Pinelliae Rhizoma Praeparatum cum Alumine; group 9: Poria and Glycyrrhizae Radix et Rhizoma Praeparata cum Melle. The time-lapse analysis showed that the proportion of drugs used in the past three years such as Glycyrrhizae Radix et Rhizoma Praeparata cum Melle, Poria and Aurantii Fructus was gradually increasing.Conclusions:Professor Li's treatment of cough focuses on the lungs, spleen, and stomach. Clinical medication emphasizes the combination of ascending and descending factors, as well as the use of cold and warm. In recent years, there has been a greater emphasis on treating cough from the middle energizer.

As a kind of traditional Chinese medicine,which has a medicinal site of rhizomes,contains rich active ingredients,mainly including steroidal saponins,flavonoids,phenols,stilbenes,organic acids and other compounds,which together give a wide range of pharmacological effects,including anti-inflammatory,antibacterial,anti-tumor,hypolipidemic,hypoglycemic,and so on.In recent years,with the continuous development of modern science and technology,the research on Smilacis chinae rhizoma has been deepening,and the pharmacological mechanism of its active ingredients has been gradually revealed.This paper reviews the research progress on the active ingredients and pharmacological effects of Smilacis chinae rhizoma over the last few years at home and abroad,in order to provide a scientific basis for further development and utilization of Smilacis chinae rhizoma,and inject new vitality into the modernization and international development of traditional Chinese medicine.

Objective To establish the method for extracting exogenous short DNA fragments of Schistosoma japonicum from urine samples, and to evaluate the efficiency of this method for extraction from urine samples treated with various methods. Methods The S. japonicum SjG28 gene fragment was selected as a target sequence, and the 81 bp short DNA fragment was amplified on the target sequence using PCR assay. Following characterization using sequencing, the short DNA fragment was added into the urine samples as an exogenous short DNA fragment. Primers and probes were designed with SjG28 as a target gene, to establish the real-time fluorescent quantitative PCR (qPCR) assay. The sensitivity of this qPCR assay was evaluated with exogenous short DNA fragments that were diluted at a 1:10 dilution ratio as the DNA template, and the specificity of the qPCR assay was evaluated with the genomic DNA of S. mansoni, S. haematobium, Babesia, Ancyiostoma duodenaie, Cionorchis sinensis, and Paragonimus westermani as DNA templates. Exogenous short DNA fragments were added into artificial and healthy volunteers’ urine samples, followed by pH adjustment, centrifugation and concentration, and the efficiency of extracting exogenous short DNA fragments from urine samples was compared with the QIAmp Viral RNA Mini Kit (Qiagen kit) and BIOG cfDNA easy kit (BIOG kit). Results An 81 bp small DNA fragment of S. japonicum was successfully prepared, and the lowest detection limit of the established qPCR assay was 100 copies/μL of the 81 bp small DNA fragment of S. japonicum. If the genomic DNA of S. japonicum, S. mansoni, S. haematobium, Babesia, A. duodenaie, C. sinensis, and P. westermani served as DNA templates, the qPCR assay only detected fluorescent signals with S. japonicum genomic DNA as the DNA template. If the pH values of artificial urine samples were adjusted to 5, 6, 7 and 8, the recovery rates were (49.12 ± 2.09)%, (84.52 ± 4.96)%, (89.38 ± 3.32)% and (87.82 ± 3.90)% for extracting the exogenous short DNA fragment of S. japonicum with the Qiagen kit, and were (2.30 ± 0.07)%, (8.11% ± 0.26)%, (13.35 ± 0.61)% and (20.82 ± 0.68)% with the BIOG kit, respectively (t = 38.702, 26.955, 39.042 and 29.571; all P values < 0.01). If the Qiagen kit was used for extracting the exogenous short DNA fragment from artificial urine samples, the lowest recovery rate was seen from urine samples with a pH value of 5 (all P values < 0.05), and there were no significant differences in the recovery rate from urine samples with pH values of 6, 7 and 8 (all P values > 0.05). Following centrifugation of artificial [(64.30 ± 1.00)% vs. (58.87 ± 0.26)%; t = 12.033, P < 0.05] and healthy volunteers’ urine samples [(31 165 ± 1 017) copies/μL vs. (28 471 ± 818) copies/μL; t = 23.164, P < 0.05]. In addition, concentration of artificial urine samples with the 10 kDa Centrifugal Filter and concentration of healthy volunteers’ urine samples with the 100 kDa Centrifugal Filter were both effective to increase the recovery of the Qiagen kit for extracting the exogenous short DNA fragment of S. japonicum (both P values < 0.01). Conclusions A method for extracting exogenous short DNA fragments of S. japonicum from urine samples has been successfully established, and the Qiagen kit has a high extraction efficiency. Adjustment of urine pH to 6 to 8 and concentration of healthy volunteers’ urine samples with the 100 kDa Centrifugal Filter are both effective to increase the efficiency of extracting exogenous short DNA fragments of S. japonicum.

Guillain-Barré syndrome (GBS) defines a kind of Immune-mediated acute inflammatory peripheral neuropathy. Miller-Fisher Syndrome (MFS) is a special variant of GBS, with mostly one-way course and rare clinical recurrence. Only a few recurrent cases have been reported in China. Here we report a case of a young male patient with double vision and progressive aggravation of limb numbness, acute onset, with symptoms of upper respiratory tract infection before onset, accompanied by pupil abnormalities and autonomic nervous dysfunction, who was was admitted to our hospital for similar symptoms 3 years ago and was improved by immunotherapy. The patient had a triad of “ataxia, areflexia and ophthalmoplegia”. Cerebrospinal fluid showed protein-cell separation. Serum anti-Sulfatides antibody IgM, anti-GT1a antibody IgG, anti-GQ1b antibody IgG and anti-GM3 IgM were positive. Recurrent MFS was diagnosed and the symptoms improved after immunotherapy. This case suggests that MFS is clinically heterogeneous, a few patients can present with relapse and generally have a better prognosis with immunotherapy. Pre-existing infection and anti-GQ1b antibody production may be predisposing factors for MFS recurrence.