Functional dyspepsia （FD） is a prioritized disease category where traditional Chinese medicine （TCM） demonstrates distinct therapeutic advantages. The current western medicine treatment for FD is mainly based on proton pump inhibitors and prokinetic agents， with digestive enzymes， probiotics and antidepressants serving as adjuvant medication， yet such therapies still have certain limitations. TCM treatment for FD includes oral administration of Chinese herbal formulas and Chinese patent medicines， as well as external TCM therapies such as acupuncture and moxibustion， acupoint application， hot medicinal compress therapy， rubbing with ointment， medicinal iontophoresis， auricular acupoint therapy and tui na （Chinese medical massage）. The combined treatment of FD with integrated TCM and western medicine can significantly improve clinical effectiveness and reduce adverse reactions. The common mechanisms underlying the therapeutic effects of both TCM and western medicine revolve around the core pathological processes of FD， mainly focusing on restoring gastrointestinal motility， regulating the levels of brain-gut peptides， modulating intestinal microecology， and ameliorating inflammatory status. The differential mechanisms lie in the precise targeting feature of western medicine versus the holistic-regulating and multi-target characteristics of TCM， and the two approaches exert a synergistic effect to enhance efficacy. This paper proposes to leverage the advantages of TCM in holistic regulation and the strengths of western medicine in targeted treatment， so as to provide personalized and comprehensive treatment regimens for FD patients.

OBJECTIVE To investigate the effects and mechanism of the Yishen paidu formula on renal fibrosis in rats with chronic renal failure （CRF） through the reactive oxygen species （ROS）/thioredoxin-interacting protein （TXNIP）/NOD-like receptor thermal protein domain associated protein 3 （NLRP3） pathway. METHODS Rats were randomly divided into control group， model group， Yishen paidu formula low-dose （Yishen paidu formula-L） group， Yishen paidu formula high-dose （Yishen paidu formula- H） group， Yishen paidu formula-H+pcDNA-NC group， and Yishen paidu formula-H+ pcDNA-TXNIP group， with 10 rats in each group. Except for control group， all other rats were fed a diet containing 0.5% adenine to establish a CRF model； the rats were then administered corresponding drugs or normal saline intragastrically or via tail vein， once daily， for 8 consecutive weeks. After the last administration， the levels of serum creatinine （Scr）， blood urea nitrogen （BUN）， ROS， superoxide dismutase （SOD）， malondialdehyde （MDA）， tumor necrosis factor-α （TNF-α）， interleukin （IL）-6， and IL-1β were measured in each group. Pathological changes in renal tissue were observed， and the protein expression levels of Collagen Ⅲ， α-smooth muscle actin （α-SMA）， transforming growth factor-β1 （TGF-β1）， TXNIP and NLRP3 in renal tissue were detected. RESULTS Compared with model group， the renal histopathological damage and fibrosis of rats in Yishen paidu formula-L group and Yishen paidu formula-H group were significantly alleviated. The levels of Scr， BUN， ROS， MDA， TNF- α， IL-6 and IL-1β， and the protein expressions of Collagen Ⅲ， α-SMA， TGF-β1， TXNIP and NLRP3 were significantly decreased， while SOD levels were significantly increased （P＜0.05）. Moreover， the changes were more pronounced in the Yishen paidu formula-H group （P＜0.05）. Compared with Yishen paidu formula-H+pcDNA-NC group， above indexes of rats in Yishen paidu formula-H+pcDNA-TXNIP group were reversed significantly （P＜0.05）. CONCLUSIONS Yishen paidu formula can inhibit renal fibrosis in CRF rats by suppressing the ROS/TXNIP/NLRP3 pathway.

OBJECTIVE: To investigate the effectiveness and preliminary mechanisms of icariin (ICA) in enhancing the reparative effects of adipose-derived stem cells (ADSCs) on skin radiation damagies in rats. METHODS: Twelve SPF-grade Sprague Dawley rats [body weight (220±10) g] were subjected to a single dose of 10 Gy X-ray irradiation on a 1.5 cm×1.5 cm area of their dorsal skin, with a dose rate of 200 cGy/min to make skin radiation damage model. After successful modelling, the rats were randomly divided into 4 groups ( n=3), and on day 2, the corresponding cells were injected subcutaneously into the irradiated wounds: group A received 0.1 mL of rat ADSCs (1×10 ⁷cells/mL), group B received 0.1 mL of rat ADSCs (1×10 ⁷cells/mL)+1 μmol/L ICA (0.1 mL), group C received 0.1 mL of rat ADSCs (1×10 ⁷cells/mL) pretreated with a hypoxia-inducible factor 2α (HIF-2α) inhibitor+1 μmol/L ICA (0.1 mL), and group D received 0.1 mL of rat ADSCs (1×10 ⁷cells/mL) pretreated with a Notch1 inhibitor+1 μmol/L ICA (0.1 mL). All treatments were administered as single doses. The skin injury in the irradiated areas of the rats was observed continuously from day 1 to day 7 after modelling. On day 28, the rats were sacrificed, and skin tissues from the irradiated areas were harvested for histological examination (HE staining and Masson staining) to assess the repair status and for quantitative collagen content detection. Immunohistochemical staining was performed to detect CD31 expression, while Western blot and real-time fluorescence quantitative PCR (qRT-PCR) were used to measure the protein and mRNA relative expression levels of vascular endothelial growth factor (VEGF), platelet-derived growth factor BB (PDGF-BB), fibroblast growth factor 2 (FGF-2), interleukin 10 (IL-10), transforming growth factor β (TGF-β), HIF-2α, and Notch1, 2, and 3. RESULTS: All groups exhibited skin ulcers and redness after irradiation. On day 3, exudation of tissue fluid was observed in all groups. On day 7, group B showed significantly smaller skin injury areas compared to the other 3 groups. On day 28, histological examination revealed that the epidermis was thickened and the dermal fibers were slightly disordered with occasional inflammatory cell aggregation in group A. In group B, the epidermis appeared more normal, the dermal fibers were more orderly, and there was an increase in new blood vessels without significant inflammatory cell aggregation. In contrast, groups C and D showed significantly increased epidermal thickness, disordered and disrupted dermal fibers. Group B had higher collagen fiber content than the other 3 groups, and group D had lower content than group A, with significant differences ( P<0.05). Immunohistochemical staining showed that group B had significantly higher CD31 expression than the other 3 groups, while groups C and D had lower expression than group A, with significant differences ( P<0.05). Western blot and qRT-PCR results indicated that group B had significantly higher relative expression levels of VEGF, PDGF-BB, FGF-2, IL-10, TGF-β, HIF-2α, and Notch1, 2, and 3 proteins and mRNAs compared to the other 3 groups ( P<0.05). CONCLUSION ICA may enhance the reparative effects of ADSCs on rat skin radiation damage by promoting angiogenesis and reducing inflammatory responses through the HIF-2α-VEGF-Notch signaling pathway.
Animals ; Rats, Sprague-Dawley ; Skin/pathology* ; Rats ; Vascular Endothelial Growth Factor A/genetics* ; Basic Helix-Loop-Helix Transcription Factors/genetics* ; Signal Transduction ; Flavonoids/pharmacology* ; Adipose Tissue/cytology* ; Stem Cells/cytology* ; Receptors, Notch/metabolism* ; Radiation Injuries, Experimental/metabolism* ; Wound Healing/drug effects* ; Male

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 compare the accuracy and applicability of three adult height prediction methods based on artificial intelligence-assisted bone age assessment—the Bayley-Pinneau method(BP method), the Tanner-Whitehouse 3 method(TW3 method), and China 05 method—in girls.Methods:This bidirectional cohort study collected clinical data and 690 posteroanterior X-ray images of the left hand from 278 female children who underwent pubertal development assessments at Shenzhen Children′s Hospital between January 2014 and December 2020, with follow-up until adult height was reached. Adult height prediction was performed using BP, TW3, and China 05 methods on artificial intelligence-assisted bone age assessment.Results:The BP and TW3 methods overestimated adult height by(1.7±3.7) cm and(2.6±3.0) cm, respectively, while the China 05 method underestimated adult height by(2.3±3.5) cm. The proportion of PAH within±5 cm of FAH were 80.0% for the TW3 method, 77.0% for the BP method, and 74.2% for the China 05 method, with significant differences among them( P=0.038). Analysis of cases with prediction deviations>10 cm and subgroup comparisons revealed that the TW3 and BP methods were more likely to overestimate adult height in girls aged 6.0-<8.0 years, with delayed bone age, or in the prepubertal stage(all P<0.001). The China 05 method was more prone to underestimate adult height in those with advanced bone age( P<0.001). All three methods showed significantly greater prediction errors(absolute difference between PAH and FAH) in girls with early puberty compared to those with normal pubertal development(all P<0.05). Conclusions:The TW3 and BP methods tend to overestimate adult height in girls, while the China 05 method tends to underestimate it. Caution is warranted when predicting adult height, particularly in girls under 8 years of bone age, with delayed or advanced bone age, and those with early puberty.

Objective:To explore effect of Yishen Paidu Formula on inflammatory response in chronic renal failure(CRF)rats and role of Calcineurin/nuclear factor of activated T cell(NFAT).Methods:CRF rat model was constructed,and randomly grouped into model group,low,medium and high doses Yishen Paidu Formula groups,with 10 rats in each group,another 10 rats were fed normally as a blank group;general situation of rats was recorded;urine protein quantification kit was applied to detect 24-hour urine protein level of CRF rats in each group;serum creatinine and urea nitrogen levels of CRF rats were detected;ELISA was applied to detect serum IL-1β,IL-6 and TNF-α levels in CRF rats;pathological changes of renal tissue were observed by HE staining;Western blot was applied to detect expressions of fibroblast growth factor 23(FGF23),Klotho protein,Calcineurin,NFAT protein in renal tissue of CRF rats in each group.Results:Compared with blank group,levels of creatinine,urea nitrogen in serum,24 h urine protein in urine,IL-1β,IL-6,TNF-α in serum,and protein levels of FGF23,Calcineurin,NFAT in kidney tissue were obviously increased in model group,level of Klotho protein was obviously decreased(P<0.05).Compared with model group,levels of creatinine,urea nitrogen in serum,24 h urine protein in urine,IL-1β,IL-6,TNF-α in serum,and protein levels of FGF23,Calcineurin,NFAT in kidney tissue were obviously decreased in low,medium and high doses Yishen Paidu Formula groups,level of Klotho protein was obviously increased,which were more significant with dosage increase(P<0.05).Conclusion:Yishen Paidu Formula may alleviate inflammatory response in CRF rats by inhibiting Calcineurin/NFAT signaling pathway.

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 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.

Objective:To explore effect of Yishen Paidu Formula on inflammatory response in chronic renal failure(CRF)rats and role of Calcineurin/nuclear factor of activated T cell(NFAT).Methods:CRF rat model was constructed,and randomly grouped into model group,low,medium and high doses Yishen Paidu Formula groups,with 10 rats in each group,another 10 rats were fed normally as a blank group;general situation of rats was recorded;urine protein quantification kit was applied to detect 24-hour urine protein level of CRF rats in each group;serum creatinine and urea nitrogen levels of CRF rats were detected;ELISA was applied to detect serum IL-1β,IL-6 and TNF-α levels in CRF rats;pathological changes of renal tissue were observed by HE staining;Western blot was applied to detect expressions of fibroblast growth factor 23(FGF23),Klotho protein,Calcineurin,NFAT protein in renal tissue of CRF rats in each group.Results:Compared with blank group,levels of creatinine,urea nitrogen in serum,24 h urine protein in urine,IL-1β,IL-6,TNF-α in serum,and protein levels of FGF23,Calcineurin,NFAT in kidney tissue were obviously increased in model group,level of Klotho protein was obviously decreased(P<0.05).Compared with model group,levels of creatinine,urea nitrogen in serum,24 h urine protein in urine,IL-1β,IL-6,TNF-α in serum,and protein levels of FGF23,Calcineurin,NFAT in kidney tissue were obviously decreased in low,medium and high doses Yishen Paidu Formula groups,level of Klotho protein was obviously increased,which were more significant with dosage increase(P<0.05).Conclusion:Yishen Paidu Formula may alleviate inflammatory response in CRF rats by inhibiting Calcineurin/NFAT signaling pathway.