This study aims to explore the effects and action mechanisms of the active ingredients in Buyang Huanwu Decoction(BYHWD), namely tetramethylpyrazine(TMP) and hydroxy-safflor yellow A(HSYA), on oxygen-glucose deprivation/reglucose-reoxygenation(OGD/R)-induced inflammation and oxidative stress of microglia(MG). Network pharmacology was used to screen the effective monomer ingredients of BYHWD and determine the safe concentration range for each component. Inflammation and oxidative stress models were established to further screen the best ingredient combination and optimal concentration ratio with the most effective anti-inflammatory and antioxidant effects. OGD/R BV2 cell models were constructed, and BV2 cells in the logarithmic growth phase were divided into a normal group, a model group, an HSYA group, a TMP group, and an HSYA + TMP group. Enzyme-linked immunosorbent assay(ELISA) was used to detect the levels of inflammatory cytokines such as interleukin-1β(IL-1β), tumor necrosis factor-α(TNF-α), and interleukin-6(IL-6). Oxidative stress markers, including superoxide dismutase(SOD), nitric oxide(NO), and malondialdehyde(MDA), were also measured. Western blot was used to analyze the protein expression of both inflammation-related pathway [Toll-like receptor 4(TLR4)/nuclear factor-kappa B(NF-κB)] and oxidative stress-related pathway [nuclear factor erythroid 2-related factor 2(Nrf2)/heme oxygenase-1(HO-1)]. Immunofluorescence was used to assess the expression of proteins such as inducible nitric oxide synthase(iNOS) and arginase-1(Arg-1). The most effective ingredients for anti-inflammatory and antioxidant effects in BYHWD were TMP and HSYA. Compared to the normal group, the model group showed significantly increased levels of IL-1β, TNF-α, IL-6, NO, and MDA, along with significantly higher protein expression of NF-κB, TLR4, Nrf2, and HO-1 and significantly lower SOD levels. The differences between the two groups were statistically significant. Compared to the model group, both the HSYA group and the TMP group showed significantly reduced levels of IL-1β, TNF-α, IL-6, NO, and MDA, lower expression of NF-κB and TLR4 proteins, higher levels of SOD, and significantly increased protein expression of Nrf2 and HO-1. Additionally, the expression of the M1-type MG marker iNOS was significantly reduced, while the expression of the M2-type MG marker Arg-1 was significantly increased. The results of the HSYA group and the TMP group had statistically significant differences from those of the model group. Compared to the HSYA group and the TMP group, the HSYA + TMP group showed further significant reductions in IL-1β, TNF-α, IL-6, NO, and MDA levels, along with significant reductions in NF-κB and TLR4 protein expression, an increase in SOD levels, and elevated Nrf2 and HO-1 protein expression. Additionally, the expression of the M1-type MG marker iNOS was reduced, while the M2-type MG marker Arg-1 expression increased significantly in the HSYA + TMP group compared to the TMP or HSYA group. The differences in the results were statistically significant between the HSYA + TMP group and the TMP or HSYA group. The findings indicated that the combined use of HSYA and TMP, the active ingredients of BYHWD, can effectively inhibit OGD/R-induced inflammation and oxidative stress of MG, showing superior effects compared to the individual use of either component.
Oxidative Stress/drug effects* ; Drugs, Chinese Herbal/pharmacology* ; Animals ; Mice ; Glucose/metabolism* ; Cell Line ; Inflammation/genetics* ; Oxygen/metabolism* ; Pyrazines/pharmacology* ; Microglia/metabolism* ; NF-E2-Related Factor 2/immunology* ; NF-kappa B/immunology* ; Toll-Like Receptor 4/immunology* ; Anti-Inflammatory Agents/pharmacology* ; Humans

Objective:To analyze the application efficacy of employing high-density porous polyethylene （Su-por） in combination with temporoparietal fascial flaps via a minimally invasive scalp incision in auricular reconstruction. Methods:This study carried out a retrospective analysis of 50 patients （50 ears in total） who underwentprimary auricular reconstruction with a Su-por scaffold in our hospital from June 2022 to January 2024. All patients underwent primary auricular reconstruction using a minimally invasive scalp incision with high-density porous polyethylene （Su-por） and temporoparietal fascial flaps. The postoperative treatment effects and complications were statistically analyzed. Results:The reconstructed ears of all patients survived. After 6 months of follow-up, the scar hyperplasia of the scalp minimally invasive incision was not obvious in any patient, and no significant hair loss was observed. The reconstructed auricle of 48 patients had a realistic shape and strong three-dimensional sense. With the extension of follow-up time, the three-dimensional structure of the auricle became clearer, and patient satisfaction increased. Among the remaining two patients, one case of flap necrosis survived after skin grafting and dressing changes. One patient had scar hyperplasia at the incision of the reconstructed ear due to a scar-prone constitution, and the shape of the auricle was not ideal, but the scar hyperplasia at the scalp incision was not obvious. Conclusion:One-stage ear reconstruction with high-density porous polyethylene （Su-por） combined with superficial temporal fascia flap through a minimally invasive scalp incision can better show the fine structure of the reconstructed ear. The minimally invasive scalp incision can effectively reduce the occurrence of scar hyperplasia and postoperative alopecia at the scalp incision.
Humans ; Plastic Surgery Procedures/methods* ; Retrospective Studies ; Surgical Flaps ; Tissue Scaffolds ; Polyethylene ; Ear Auricle/surgery* ; Male ; Scalp/surgery* ; Female ; Skin Transplantation ; Fascia/transplantation* ; Porosity ; Adult ; Middle Aged

ObjectiveTo investigate the noise level and influencing factors in metro platforms and station halls， thereby providing the scientific basis for the establishment of hygienic standards. MethodsDuring the morning peak（7：00‒9：30）and off-peak （9：30‒17：00） on weekdays， the noise levels were measured with noise meters at 39 monitoring points of 13 station platforms and 31 monitoring points of 6 station halls. The monitoring points arrangement and detection methods referred to the Examination methods for public places—Part 1： physical parameters（GB/T 18204.1‒2013）. ResultsThe measured noise level in the station ranged from 69.25 to 86.17 dB（A）， accounting for 44.74% below 75 dB（A）， 89.47% below 80 dB（A） and 97.37% below 85 dB（A）.The noise level of the platform ［（76.38±4.19） dB（A）］ was higher than that of the station hall ［（74.24±4.50） dB（A）］（P<0.01）. The noise level of the elevated platforms ［（80.01±2.25） dB（A）］ was higher than that of the underground platforms ［（75.73±4.13） dB（A）］（P<0.01）， and the noise level of the platforms without platform screen doors（PSD） ［（80.21±5.08） dB（A）］ was higher than that of platforms with PSD［（74.73±3.16） dB（A）］ （P<0.01）. No statistical significant differences were observed among the different areas of the platforms， monitoring periods， platform depth， exit mode and operation years （P>0.05）. ConclusionThe noise level in metro stations in the city does not fully meet the requirements of current relevant standards. It is suggested to take noise reduction measures to reduce the noise of metro stations.

OBJECTIVE To explore the clinical effect of concurrent chemoradiotherapy combined with nituzumab in the treatment of locally advanced nasopharyngeal carcinoma in Guangxi,Yunnan and Guizhou.METHODS A total of 80 patients with locally advanced nasopharyngeal carcinoma who were pathologically confirmed and admitted to Affiliated Hospital of Youjiang Ethnomedicine from July 2021 to July 2022 from ethnic minority areas near the border of Guangxi,Yunnan and Guizhou were selected as the subjects of this study.They were randomly divided into control group(standard concurrent chemoradiotherapy)and observation group(combined treatment with nituzumab on the basis of control group),with 40 cases in each group.The levels of tumor markers,oxidative stress indicators,adverse reactions,complications,and recent clinical efficacy of the two groups were compared.RESULTS After treatment,the levels of superoxide dismutase(SOD),cytokeratin 19 fragment antigen21-1(CYFRA21-1),squamous cell carcinoma associated antigen(SCCAg)and serum ferritin(SF)were significantly decreased in both groups,while nitric oxide(NO)and malondialdehyde(MDA)were significantly increased.The levels of SCCAg,SF,CYFRA21-1,NO and MDA in observation group were lower than those in control group,and the level of SOD was higher than those in control group(P<0.05).Compared with the control group,the objective response rate(ORR)in the observation group was increased(P<0.05).There was no significant difference in the incidence of adverse reactions between the two groups(P>0.05).There was no significant difference in the incidence of complications between the two groups(P>0.05).CONCLUSION Concurrent chemoradiotherapy combined with nituzumab can effectively improve the short-term survival rate and clinical efficacy of patients with locally advanced nasopharyngeal carcinoma,regulate tumor markers and oxidative stress levels,and alleviate the disease.

Background::Total human immunodeficiency virus (HIV) DNA and integrated HIV DNA are widely used markers of HIV persistence. Droplet digital polymerase chain reaction (ddPCR) can be used for absolute quantification without needing a standard curve. Here, we developed duplex ddPCR assays to detect and quantify total HIV DNA and integrated HIV DNA.Methods::The limit of detection, dynamic ranges, sensitivity, and reproducibility were evaluated by plasmid constructs containing both the HIV long terminal repeat (LTR) and human CD3 gene (for total HIV DNA) and ACH-2 cells (for integrated HIV DNA). Forty-two cases on stable suppressive antiretroviral therapy (ART) were assayed in total HIV DNA and integrated HIV DNA. Correlation coefficient analysis was performed on the data related to DNA copies and cluster of differentiation 4 positive (CD4 +) T-cell counts, CD8 + T-cell counts and CD4/CD8 T-cell ratio, respectively. The assay linear dynamic range and lower limit of detection (LLOD) were also assessed. Results::The assay could detect the presence of HIV-1 copies 100% at concentrations of 6.3 copies/reaction, and the estimated LLOD of the ddPCR assay was 4.4 HIV DNA copies/reaction (95% confidence intervals [CI]: 3.6-6.5 copies/reaction) with linearity over a 5-log 10-unit range in total HIV DNA assay. For the integrated HIV DNA assay, the LLOD was 8.0 copies/reaction (95% CI: 5.8-16.6 copies/reaction) with linearity over a 3-log 10-unit range. Total HIV DNA in CD4 + T cells was positively associated with integrated HIV DNA ( r = 0.76, P <0.0001). Meanwhile, both total HIV DNA and integrated HIV DNA in CD4 + T cells were inversely correlated with the ratio of CD4/CD8 but positively correlated with the CD8 + T-cell counts. Conclusions::This ddPCR assay can quantify total HIV DNA and integrated HIV DNA efficiently with robustness and sensitivity. It can be readily adapted for measuring HIV DNA with non-B clades, and it could be beneficial for testing in clinical trials.

Celastrol and wilforlide A are the main active triterpenoids of the traditional Chinese medicine Lei Gong Teng, which have anti-tumour, anti-inflammatory and immunosuppressive activities, and are the material basis for the clinical efficacy of Lei Gong Teng-related Chinese medicinal preparations. By analysing the biosynthetic pathway of active ingredients, optimizing genetic elements and utilizing "cell factory" to produce triterpenoids heterologously will be an effective way to obtain from Tripterygium wilfordii in the future in a "low-cost and high-efficient" manner. CYP712Ks are the first cytochrome P450s involved in the skeleton modification of friedelane-type and oleanane-type triterpenoids in T. wilfordii, and they can catalyse the generation of polpunonic acid from friedelin and 3-epi-katonic acid from β-amyrin by carboxylation at C-29 position. In this study, four multifunctional TwCYP712K1/2/3/5 were used to clarify the catalytic function and substrate selection preference using in vivo functional characterization in Saccharomyces cerevisiae. The spatial structure of the protein-substrate binding was clarified through homology modeling and molecular docking, and then the differential amino acids in the active pocket of the protein were mutated to clarify the crucial amino acids determining the catalytic function and the selection of substrate structure. A total of 63 mutant elements were constructed, and the amino acid sites affecting the carboxylation function of TwCYP712Ks were analyzed. In particular, the key amino acids affecting the substrate selectivity of TwCYP712K2 towards oleanane-type and friedelane-type triterpenoids were revealed and the TwCYP712K2^F127I and TwCYP712K2^A227T mutants would result in a reversal of the product ratio. In conclusion, four proteins of TwCYP712Ks were semi-rationally designed by homologous protein alignment and mutual mutation to elucidate multiple amino acid sites determining the catalytic function of the proteins, and a series of activity-enhancing or altering mutants were obtained, which provide abundant catalytic elements for the biosynthesis of active triterpenoids from T. wilfordii, and the mechanism of carboxylation in the C-29 position was initially elucidated.

[Objective]To investigate the impact of varying concentrations of baicalein on the proliferation and biological responses of MC3T3-E1 cells,as well as the antibacterial efficacy of baicalein against prevalent oral bacteria,and to elucidate the underlying mechanisms.[Methods]MC3T3-E1 cells were exposed to different concentrations of baicalein(0,6,12,18,and 24 μmol/L)and cell viability was determined by using the CCK-8 assay.Alkaline phosphatase(ALP)activity of MC3T3-E1 cells following osteogenic induction was assessed.RT-PCR was used to examine the expression of RunX2,BMP2,and Osterix.After 24 hours of treatment,the antibacterial potential of baicalein against Escherichia coli,Staphylococcus Aureus and Streptococcus Sanguis was evaluated by using the K-B paper disk method.[Results]Baicalein exhibited a modest reduction in proliferation of MC3T3-E1 cells but without affecting their sustained proliferation.Baicalein at a concentration of 18 μmol/L enhanced ALP activity of MC3T3-E1 cells,upregulated BMP2 and Osterix expression,downregulated RunX2 expression,significantly inhibited the proliferation of Staphylococcus Aureus and Streptococcus Sanguis(P<0.05).[Conclusions]Baicalein at an optimal concentration(18 μmol/L)demonstrated a promotional effect on the osteogenic differentiation of MC3T3-E1 cells and effectively suppressed the proliferation of common oral bacteria,including Staphylococcus Aureus and Streptococcus Sanguis.

Objective To monitor the susceptibility of clinical isolates to antimicrobial agents in tertiary hospitals in major regions of China in 2022.Methods Clinical isolates from 58 hospitals in China were tested for antimicrobial susceptibility using a unified protocol based on disc diffusion method or automated testing systems.Results were interpreted using the 2022 Clinical &Laboratory Standards Institute(CLSI)breakpoints.Results A total of 318 013 clinical isolates were collected from January 1,2022 to December 31,2022,of which 29.5％were gram-positive and 70.5％were gram-negative.The prevalence of methicillin-resistant strains in Staphylococcus aureus,Staphylococcus epidermidis and other coagulase-negative Staphylococcus species(excluding Staphylococcus pseudintermedius and Staphylococcus schleiferi)was 28.3％,76.7％and 77.9％,respectively.Overall,94.0％of MRSA strains were susceptible to trimethoprim-sulfamethoxazole and 90.8％of MRSE strains were susceptible to rifampicin.No vancomycin-resistant strains were found.Enterococcus faecalis showed significantly lower resistance rates to most antimicrobial agents tested than Enterococcus faecium.A few vancomycin-resistant strains were identified in both E.faecalis and E.faecium.The prevalence of penicillin-susceptible Streptococcus pneumoniae was 94.2％in the isolates from children and 95.7％in the isolates from adults.The resistance rate to carbapenems was lower than 13.1％in most Enterobacterales species except for Klebsiella,21.7％-23.1％of which were resistant to carbapenems.Most Enterobacterales isolates were highly susceptible to tigecycline,colistin and polymyxin B,with resistance rates ranging from 0.1％to 13.3％.The prevalence of meropenem-resistant strains decreased from 23.5％in 2019 to 18.0％in 2022 in Pseudomonas aeruginosa,and decreased from 79.0％in 2019 to 72.5％in 2022 in Acinetobacter baumannii.Conclusions The resistance of clinical isolates to the commonly used antimicrobial agents is still increasing in tertiary hospitals.However,the prevalence of important carbapenem-resistant organisms such as carbapenem-resistant K.pneumoniae,P.aeruginosa,and A.baumannii showed a downward trend in recent years.This finding suggests that the strategy of combining antimicrobial resistance surveillance with multidisciplinary concerted action works well in curbing the spread of resistant bacteria.

Primary intestinal diffuse large B-cell lymphoma(PI-DLBCL)is clinically rare,but in recent years,with the gradual maturity of pathology and molecular biology technology,its incidence rate and diagnosis rate have also increased.Due to the lack of specificity of the clinical symptoms of PI-DLBCL,it is easy to misdiagnose and miss the diagnosis,and there is no consensus on the best treatment of PI-DLBCL in clinical practice.Therefore,by retrieving the latest literature at home and abroad,this review systematically discusses the pathogenesis,clinical manifestations,diagnostic criteria,treatment and prognosis of PI-DLBCL,in order to improve the understanding of rare PI-DLBCL in hematology and oncology,and provide reference for basic research and clinical diagnosis and treatment of PI-DLBCL.