In recent decades,the incidence of human papillomavirus(HPV)-associated oropharyngeal squamous cell carcinoma(OPSCC)has shown a marked increase.Significant changes have also occurred in the OPSCC diagnosis and treatment paradigm.Deter-mining HPV status prior to treatment is now essential,and radiotherapy/chemotherapy,immunotherapy,and minimally invasive surgical techniques have progressively emerged as key modalities for managing OPSCC.However,alongside these paradigm shifts,a comprehen-sive technical consensus guiding the entire diagnostic and therapeutic process for OPSCC patients is currently lacking.Given China's large population base and the rising incidence of OPSCC,an expert panel convened to develop a clinical technical consensus on OPSCC diagno-sis and management tailored to China's specific context.This consensus aims to further enhance and standardize understanding of OPSCC management techniques among relevant healthcare professionals.

Objective:To investigate the effects of baicalin on ferroptosis of mouse fibroblasts (Fbs) under high glucose treatment and its mechanism, and to provide a basis for the treatment of diabetic wounds.Methods:The study was an experimental study. Mouse Fbs were collected and divided into control group with conventional culture, high glucose group treated with glucose at final molarity of 30.0 mmol/L, and low baicalin group and high baicalin group pretreated with baicalin at final molarties of 5 and 10 μmol/L respectively and then treated as that in high glucose group. After 48 h of culture, the cell survival rate was detected by the cell counting kit-8, the reactive oxygen species level in cells was detected by the fluorescent probe method, the levels of malondialdehyde, glutathione, and ferrous ion in cells were detected by colorimetry, and the protein expression levels of solute carrier family 7 member 11 (SLC7A11) and glutathione peroxidase 4 (GPX4) in cells and nuclear factor-erythroid 2-related factor 2 (Nrf2) in cytoplasm and nucleus were detected by Western blotting. Another batch of mouse Fbs were collected and divided into control group, high glucose group, high baicalin group, and high baicalin+ML385 group. The cells in the first three groups were treated as before, the cells in the last group were pretreated with baicalin and ML385 of Nrf2 inhibitor at final molarties of 10 μmol/L and then treated as that in high glucose group. After 48 h of culture, the protein expression levels of SLC7A11 and GPX4 in cells and the protein expression level of Nrf2 in cytoplasm and nucleus were detected as before. Except that the sample number in detecting SLC7A11 and GPX4 was 4, the sample number in detecting other indexes was 3.Results:After 48 h of culture, the cell survival rates in control group, high glucose group, low baicalin group, and high baicalin group were (100.0±10.7)%, (70.0±5.0)%, (80.9±3.2)%, and (91.4±1.9)%, respectively. Compared with those in control group, the cell survival rate, the glutathione level, and SLC7A11 and GPX4 protein expression levels in cells, and nuclear Nrf2 protein expression level were significantly decreased in high glucose group ( P<0.05), and the levels of reactive oxygen species, malondialdehyde, and ferrous ion in cells, and cytoplasmic Nrf2 protein expression level were significantly increased in high glucose group ( P<0.05). Compared with those in high glucose group, the cell survival rate, glutathione level, SLC7A11 and GPX4 protein expression levels in cells, and nuclear Nrf2 protein expression level in low baicalin group and high baicalin group were significantly increased ( P<0.05), the reactive oxygen species and ferrous ion levels in cells, and cytoplasmic Nrf2 protein expression level in low baicalin group and high baicalin group were significantly decreased ( P<0.05), and the malondialdehyde level in cells in high baicalin group was significantly decreased ( P<0.05). Compared with those in low baicalin group, the cell survival rate, glutathione level, SLC7A11 and GPX4 protein expression levels in cells, and nuclear Nrf2 protein expression level in high baicalin group were significantly increased ( P<0.05), and the reactive oxygen species, malondialdehyde, and ferrous ion levels in cells, and cytoplasmic Nrf2 protein expression level in high baicalin group were significantly decreased ( P<0.05). After 48 h of culture, compared with those in control group, the nuclear Nrf2 protein expression level and SLC7A11 and GPX4 protein expression levels in cells were significantly decreased ( P<0.05), and the cytoplasmic Nrf2 protein expression level was significantly increased in high glucose group ( P<0.05); compared with those in high glucose group, the cytoplasmic Nrf2 protein expression level was significantly decreased ( P<0.05), and the nuclear Nrf2 protein expression level and SLC7A11 and GPX4 protein expression levels in cells were significantly increased in high baicalin group ( P<0.05); compared with those in high baicalin group, the cytoplasmic Nrf2 protein expression level was significantly increased ( P<0.05), and the nuclear Nrf2 protein expression level and SLC7A11 and GPX4 protein expression levels in cells were significantly decreased in high baicalin+ML385 group ( P<0.05). Conclusions:Baicalin can inhibit the occurrence of ferroptosis in cells by activating the Nrf2 signaling pathway and up-regulating the expressions of proteins related to SLC7A11/GPX4 axis in Fbs in high glucose treatment, thus increasing the cell survival rate.

Objective:To investigate the role of the structural gene vgrG of the type Ⅵ secretion system (T6SS) of Klebsiella pneumoniae ( Kpn), and evaluate the growth ability in vitro and pathogenicity of the bacteria after vgrG was deleted. Methods:Using sequences published by the National Center for Biotechnology Information (NCBI), primers were designed to amplify the upstream and downstream homology arms of vgrG via PCR. These fragments were cloned into the vector pKO3-Km after overlapping, the recombinant vector pKO3-Km- vgrG was transferred into Kpn competent cells, and the vgrG deletion strain Δ vgrG was obtained through homologous recombination. The vgrG promoter with the complete gene fragment was amplify by PCR and cloned into the pBAD33 vector. The pBAD33- vgrG was then transferred into Δ vgrG competent cells to obtain the complemented strain CΔ vgrG. The wild-type strain (WT), Δ vgrG strain and CΔ vgrG strain were cultured in LB (Luria-Bertani) liquid medium to compare growth rates. Adhesion to human lung epithelial A549 cells and intracellular survival in macrophages Raw264.7 cells were assessed. In vivo experiments included mouse survival analysis ( n=10) and lung bacterial load quantification ( n=6). Statistical comparisons were performed using the Student t-test. Results:The Δ vgrG strain was obtained through homologous recombination. It was identified by specific primers that compared with the WT strain, the complete vgrG fragment (2 487 bp) was deleted. On this basis, the CΔ vgrG strain was obtained. Deletion of vgrG did not significantly affect Kpn growth in vitro growth ability of bacteria before on after Kpn deleted vgrG [(1.40±0.10) vs (1.20±0.30), t=0.63, P>0.05]. The viscosity of WT strain was significantly higher than that of the Δ vgrG strain [(0.96±0.04) vs (0.38±0.05), t=9.72, P<0.05], the viscosity of the CΔ vgrG strain was also significantly higher than that of the Δ vgrG strain ( P<0.05). At the cellular level, the amount of adherent bacteria of the WT strain to A549 cells was significantly greater than that of the Δ vgrG strain [(5 367.00±318.00) CFU vs (4 067.00±88.19) CFU, t=3.94, P<0.05], the amount of adherent bacteria of CΔ vgrG strain was also significantly higher than that of Δ vgrG strain ( P<0.05). After 12 h infection, the WT strain survival rate in macrophages was significantly higher than that of the Δ vgrG strain[(69.00±1.00)% vs (47.50±2.50)%, t=7.99, P<0.05]. At the animal level, the survival rate of WT strain group after lethal dose infection of mice was significantly lower than that of Δ vgrG strain group [(16.67±8.82)% vs (53.33±6.67)%, t=3.32, P<0.05]; mice infected with semi-lethal dose and the number of bacteria load in the lungs of WT strain group was significantly higher than that of the Δ vgrG strain group[ (4.97±0.06) lg CFU/g vs (4.05 ±0.04) lg CFU/g, t=12.27, P<0.01], the amount of bacteria in the lungs of mice in CΔ vgrG strain group was also significantly higher than that in Δ vgrG strain group ( P<0.01). Conclusions:The vgrG gene does not affect the growth of Kpn in vitro, but it is involved in the adhesion of Kpn to epithelial cells, resistance to macrophage killing and pathogenicity to mice.

Objective:To investigate the role of the structural gene vgrG of the type Ⅵ secretion system (T6SS) of Klebsiella pneumoniae ( Kpn), and evaluate the growth ability in vitro and pathogenicity of the bacteria after vgrG was deleted. Methods:Using sequences published by the National Center for Biotechnology Information (NCBI), primers were designed to amplify the upstream and downstream homology arms of vgrG via PCR. These fragments were cloned into the vector pKO3-Km after overlapping, the recombinant vector pKO3-Km- vgrG was transferred into Kpn competent cells, and the vgrG deletion strain Δ vgrG was obtained through homologous recombination. The vgrG promoter with the complete gene fragment was amplify by PCR and cloned into the pBAD33 vector. The pBAD33- vgrG was then transferred into Δ vgrG competent cells to obtain the complemented strain CΔ vgrG. The wild-type strain (WT), Δ vgrG strain and CΔ vgrG strain were cultured in LB (Luria-Bertani) liquid medium to compare growth rates. Adhesion to human lung epithelial A549 cells and intracellular survival in macrophages Raw264.7 cells were assessed. In vivo experiments included mouse survival analysis ( n=10) and lung bacterial load quantification ( n=6). Statistical comparisons were performed using the Student t-test. Results:The Δ vgrG strain was obtained through homologous recombination. It was identified by specific primers that compared with the WT strain, the complete vgrG fragment (2 487 bp) was deleted. On this basis, the CΔ vgrG strain was obtained. Deletion of vgrG did not significantly affect Kpn growth in vitro growth ability of bacteria before on after Kpn deleted vgrG [(1.40±0.10) vs (1.20±0.30), t=0.63, P>0.05]. The viscosity of WT strain was significantly higher than that of the Δ vgrG strain [(0.96±0.04) vs (0.38±0.05), t=9.72, P<0.05], the viscosity of the CΔ vgrG strain was also significantly higher than that of the Δ vgrG strain ( P<0.05). At the cellular level, the amount of adherent bacteria of the WT strain to A549 cells was significantly greater than that of the Δ vgrG strain [(5 367.00±318.00) CFU vs (4 067.00±88.19) CFU, t=3.94, P<0.05], the amount of adherent bacteria of CΔ vgrG strain was also significantly higher than that of Δ vgrG strain ( P<0.05). After 12 h infection, the WT strain survival rate in macrophages was significantly higher than that of the Δ vgrG strain[(69.00±1.00)% vs (47.50±2.50)%, t=7.99, P<0.05]. At the animal level, the survival rate of WT strain group after lethal dose infection of mice was significantly lower than that of Δ vgrG strain group [(16.67±8.82)% vs (53.33±6.67)%, t=3.32, P<0.05]; mice infected with semi-lethal dose and the number of bacteria load in the lungs of WT strain group was significantly higher than that of the Δ vgrG strain group[ (4.97±0.06) lg CFU/g vs (4.05 ±0.04) lg CFU/g, t=12.27, P<0.01], the amount of bacteria in the lungs of mice in CΔ vgrG strain group was also significantly higher than that in Δ vgrG strain group ( P<0.01). Conclusions:The vgrG gene does not affect the growth of Kpn in vitro, but it is involved in the adhesion of Kpn to epithelial cells, resistance to macrophage killing and pathogenicity to mice.

In recent decades,the incidence of human papillomavirus(HPV)-associated oropharyngeal squamous cell carcinoma(OPSCC)has shown a marked increase.Significant changes have also occurred in the OPSCC diagnosis and treatment paradigm.Deter-mining HPV status prior to treatment is now essential,and radiotherapy/chemotherapy,immunotherapy,and minimally invasive surgical techniques have progressively emerged as key modalities for managing OPSCC.However,alongside these paradigm shifts,a comprehen-sive technical consensus guiding the entire diagnostic and therapeutic process for OPSCC patients is currently lacking.Given China's large population base and the rising incidence of OPSCC,an expert panel convened to develop a clinical technical consensus on OPSCC diagno-sis and management tailored to China's specific context.This consensus aims to further enhance and standardize understanding of OPSCC management techniques among relevant healthcare professionals.

Objective:To investigate the effects of baicalin on ferroptosis of mouse fibroblasts (Fbs) under high glucose treatment and its mechanism, and to provide a basis for the treatment of diabetic wounds.Methods:The study was an experimental study. Mouse Fbs were collected and divided into control group with conventional culture, high glucose group treated with glucose at final molarity of 30.0 mmol/L, and low baicalin group and high baicalin group pretreated with baicalin at final molarties of 5 and 10 μmol/L respectively and then treated as that in high glucose group. After 48 h of culture, the cell survival rate was detected by the cell counting kit-8, the reactive oxygen species level in cells was detected by the fluorescent probe method, the levels of malondialdehyde, glutathione, and ferrous ion in cells were detected by colorimetry, and the protein expression levels of solute carrier family 7 member 11 (SLC7A11) and glutathione peroxidase 4 (GPX4) in cells and nuclear factor-erythroid 2-related factor 2 (Nrf2) in cytoplasm and nucleus were detected by Western blotting. Another batch of mouse Fbs were collected and divided into control group, high glucose group, high baicalin group, and high baicalin+ML385 group. The cells in the first three groups were treated as before, the cells in the last group were pretreated with baicalin and ML385 of Nrf2 inhibitor at final molarties of 10 μmol/L and then treated as that in high glucose group. After 48 h of culture, the protein expression levels of SLC7A11 and GPX4 in cells and the protein expression level of Nrf2 in cytoplasm and nucleus were detected as before. Except that the sample number in detecting SLC7A11 and GPX4 was 4, the sample number in detecting other indexes was 3.Results:After 48 h of culture, the cell survival rates in control group, high glucose group, low baicalin group, and high baicalin group were (100.0±10.7)%, (70.0±5.0)%, (80.9±3.2)%, and (91.4±1.9)%, respectively. Compared with those in control group, the cell survival rate, the glutathione level, and SLC7A11 and GPX4 protein expression levels in cells, and nuclear Nrf2 protein expression level were significantly decreased in high glucose group ( P<0.05), and the levels of reactive oxygen species, malondialdehyde, and ferrous ion in cells, and cytoplasmic Nrf2 protein expression level were significantly increased in high glucose group ( P<0.05). Compared with those in high glucose group, the cell survival rate, glutathione level, SLC7A11 and GPX4 protein expression levels in cells, and nuclear Nrf2 protein expression level in low baicalin group and high baicalin group were significantly increased ( P<0.05), the reactive oxygen species and ferrous ion levels in cells, and cytoplasmic Nrf2 protein expression level in low baicalin group and high baicalin group were significantly decreased ( P<0.05), and the malondialdehyde level in cells in high baicalin group was significantly decreased ( P<0.05). Compared with those in low baicalin group, the cell survival rate, glutathione level, SLC7A11 and GPX4 protein expression levels in cells, and nuclear Nrf2 protein expression level in high baicalin group were significantly increased ( P<0.05), and the reactive oxygen species, malondialdehyde, and ferrous ion levels in cells, and cytoplasmic Nrf2 protein expression level in high baicalin group were significantly decreased ( P<0.05). After 48 h of culture, compared with those in control group, the nuclear Nrf2 protein expression level and SLC7A11 and GPX4 protein expression levels in cells were significantly decreased ( P<0.05), and the cytoplasmic Nrf2 protein expression level was significantly increased in high glucose group ( P<0.05); compared with those in high glucose group, the cytoplasmic Nrf2 protein expression level was significantly decreased ( P<0.05), and the nuclear Nrf2 protein expression level and SLC7A11 and GPX4 protein expression levels in cells were significantly increased in high baicalin group ( P<0.05); compared with those in high baicalin group, the cytoplasmic Nrf2 protein expression level was significantly increased ( P<0.05), and the nuclear Nrf2 protein expression level and SLC7A11 and GPX4 protein expression levels in cells were significantly decreased in high baicalin+ML385 group ( P<0.05). Conclusions:Baicalin can inhibit the occurrence of ferroptosis in cells by activating the Nrf2 signaling pathway and up-regulating the expressions of proteins related to SLC7A11/GPX4 axis in Fbs in high glucose treatment, thus increasing the cell survival rate.

[Objective]To explore the therapeutic effect of Huaier(Trametes robiniophila Murr.) water extracts on granulomatous lobular mastitis(GLM) rat by reducing inflammation levels.[Methods]Female SD rats were randomly divided into normal group,model group,Huaier group[Huaier water extracts 4.5 g/(kg·d)]and hormone group[prednisone acetate 0.0018 g/(kg·d)].The GLM model was induced with tissue homogenate mixture of GLM tissue and Freund's complete adjuvant.After 14 days of administration,the mammary glands of each group were observed;the inflammation index score of the rat mammary gland was evaluated by breast inflammation index score;pathological observation of breast tissue was performed after hematoxylin-eosin(HE) staining and the level of inflammatory factors in the rat serum was detected by enzyme-linked immunosorbent assay(ELISA) method;16S rRNA amplicon sequencing technique was used to determine the diversity and richness of intestinal flora,and the relative abundance of intestinal flora in each group.[Results]Compared with normal group,the rats in model group showed symptoms such as the red breast skin,obvious lumps and significantly increased breast inflammation index score(P<0.05).Pathological changes included the formation of granulomas centered on the breast lobules,with a large number of lymphocytes,plasma cells and other inflammatory cells infiltration,which confirmed the successful induction of GLM rat model.Compared with model group,the breast swelling was improved,and the breast inflammation index score of Huaier group and hormone group both decreased(P<0.05,P<0.05).And the level of inflammatory factors interleukin-6(IL-6),interleukin-17A(IL-17A) and tumor necrosis factor-α(TNF-α) in serum significantly decreased(P<0.05,P<0.05,P<0.05).It could alleviate the infiltration of inflammatory cells in the mammary gland tissue of model group.Significant alterations in the gut microbiota of GLM rat models were observed,with statistically significant differences(P<0.05).After modeling,the abundances of Lactobacillales,Actinobacteria,Bifidobacteriales and Coriobacteriia were downregulated,while those of Caulobacterales and Desulfobacterota were upregulated within the gut microbiota.After the intervention with Huaier water extracts,the abundances of Clostridia were downregulated,while those of Bacteroidetes and Rhodospirillales were upregulated in the gut microbiota of GLM model rats.[Conclusion]Huaier water extracts could alleviate the infiltration of inflammatory cells in the mammary gland tissue of GLM model rats;reduce the proliferation of epithelial cells and the formation of inflammatory granuloma;decrease the level of inflammatory cytokines in the serum of GLM rat model and adjust the intestinal flora of GLM rat models,which may be related to its ability to improve the immunity and relieve the inflammatory response.

[Objective]To explore the therapeutic effect of Huaier(Trametes robiniophila Murr.) water extracts on granulomatous lobular mastitis(GLM) rat by reducing inflammation levels.[Methods]Female SD rats were randomly divided into normal group,model group,Huaier group[Huaier water extracts 4.5 g/(kg·d)]and hormone group[prednisone acetate 0.0018 g/(kg·d)].The GLM model was induced with tissue homogenate mixture of GLM tissue and Freund's complete adjuvant.After 14 days of administration,the mammary glands of each group were observed;the inflammation index score of the rat mammary gland was evaluated by breast inflammation index score;pathological observation of breast tissue was performed after hematoxylin-eosin(HE) staining and the level of inflammatory factors in the rat serum was detected by enzyme-linked immunosorbent assay(ELISA) method;16S rRNA amplicon sequencing technique was used to determine the diversity and richness of intestinal flora,and the relative abundance of intestinal flora in each group.[Results]Compared with normal group,the rats in model group showed symptoms such as the red breast skin,obvious lumps and significantly increased breast inflammation index score(P<0.05).Pathological changes included the formation of granulomas centered on the breast lobules,with a large number of lymphocytes,plasma cells and other inflammatory cells infiltration,which confirmed the successful induction of GLM rat model.Compared with model group,the breast swelling was improved,and the breast inflammation index score of Huaier group and hormone group both decreased(P<0.05,P<0.05).And the level of inflammatory factors interleukin-6(IL-6),interleukin-17A(IL-17A) and tumor necrosis factor-α(TNF-α) in serum significantly decreased(P<0.05,P<0.05,P<0.05).It could alleviate the infiltration of inflammatory cells in the mammary gland tissue of model group.Significant alterations in the gut microbiota of GLM rat models were observed,with statistically significant differences(P<0.05).After modeling,the abundances of Lactobacillales,Actinobacteria,Bifidobacteriales and Coriobacteriia were downregulated,while those of Caulobacterales and Desulfobacterota were upregulated within the gut microbiota.After the intervention with Huaier water extracts,the abundances of Clostridia were downregulated,while those of Bacteroidetes and Rhodospirillales were upregulated in the gut microbiota of GLM model rats.[Conclusion]Huaier water extracts could alleviate the infiltration of inflammatory cells in the mammary gland tissue of GLM model rats;reduce the proliferation of epithelial cells and the formation of inflammatory granuloma;decrease the level of inflammatory cytokines in the serum of GLM rat model and adjust the intestinal flora of GLM rat models,which may be related to its ability to improve the immunity and relieve the inflammatory response.

Objective:To construct the content framework of the rehabilitation management application (APP) for patients with urinary incontinence after radical prostatectomy, so as to provide an effective network management platform for the rehabilitation management of patients wither urinary incontinence after radical prostatectomy.Methods:From November 2020 to May 2021, the content framework of rehabilitation management APP for patients with urinary incontinence after radical prostatectomy was initially constructed through literature review and group discussion. The objective sampling was used to select 15 experts from Beijing, Shanghai and other regions for two rounds of expert consultation, to construct the content framework of rehabilitation management APP for patients with urinary incontinence after radical prostatectomy.Results:Among two rounds of expert consultation, the effective recovery rates of the questionnaires were all 100.00% (15/15) , and the expert authority coefficients were 0.937 and 0.934, respectively. The Kendall harmony coefficients of all indicators in the second round of expert consultation ranged from 0.208 to 0.333, and the differences were all statistical (all P<0.01) . It was finally determined that the content framework of rehabilitation management APP for patients with urinary incontinence after radical prostatectomy included 3 first-level indicators, 10 second-level indicators, 43 third-level indicators, and 62 fourth-level indicators. Conclusions:The content framework of constructed rehabilitation management APP for patients with urinary incontinence after radical prostatectomy is highly scientific and practical, and can provide corresponding evidence.

Objective:To explore the application value of sevoflurane inhalation anesthesia in mitral valve replacement.Methods:A total of 94 patients who underwent mitral valve replacement in our hospital (October 2016-October 2018) were randomly divided into the control group ( n=47) and the observation group ( n=47). The control group received target-controlled infusion of propofol, and the observation group inhaled sevoflurane.The postoperative conditions [intensive care unit (ICU) stay time, extubation time of tracheal tube, spontaneous cardiac rebound], hemodynamic indexes [mean arterial pressure (MAP), heart rate (HR)], serum creatine phosphokinase isoenzyme (CK-MB), cardiac troponin I (cTnI), malondialdehyde (MDA) and superoxide dismutase (SOD) in the two groups were analyzed. The patients were followed up for one month. The incidence of major adverse cardiovascular events (MACE) was calculated. Results:⑴ Postoperative situation: the time of stay in ICU and extubation of tracheal tube in the observation group was shorter than that in the control group, and the rate of spontaneous cardiac rebound (93.62%) was higher than that in the control group (72.34%) ( P<0.05); ⑵ Hemodynamic index level: there was no statistically significant difference in MAP and HR levels between two groups before operation, before cardiopulmonary bypass, after cardiopulmonary bypass, and after operation ( P>0.05); ⑶ CK-MB and cTnI: the levels of serum CK-MB and cTnI in the two groups were higher at 2, 6, 24, and 48 h after aortic cross-clamp release than before anesthesia induction, but the indicators of the observation group were lower than those in the control group; ⑷ MDA and SOD: the serum SOD level in the two groups at 2, 6, 24, and 48 h after aortic cross-clamp release were lower than before anesthesia induction, and the MDA level in the two groups at 2, 6, 24, and 48 h after aortic cross-clamp release were higher than before anesthesia induction. The level of SOD in the observation group was higher than that in the control group, and the level of MDA was lower than that in the control group ( P<0.05); ⑸ MACE: the incidence of MACE in the observation group (12.77%) was lower than that of the control group (29.79%) ( P<0.05). Conclusions:During mitral valve replacement, sevoflurane inhalation anesthesia can maintain hemodynamic stability. The duration of ICU stay and tracheal tube extubation time is shorter, and the fluctuation of serum CK-MB, cTnI, MDA and SOD is small, and it can reduce the risk of MACE.