BACKGROUND:It has been shown that Gushukang affects bone metabolism by regulating nucleotide and amino acid metabolism and immune mechanisms.Current research on the mechanism of Gushukang in the treatment of osteoporosis primarily focuses on osteoblast regulation and requires further improvement from the perspective of osteoclasts. OBJECTIVE:To investigate the mechanism by which Gushukang interferes with osteoclasts in the treatment of osteoporosis using RAW264.7 cells as the research model. METHODS:Twenty-four 8-week-old female Sprague-Dawley rats were randomly divided into four groups(n=6 per group):the three experimental groups were given 1,2 and 4 g/kg osteoporosis solution by gavage(2 times per day),and the control group was given an equal amount of distilled water by gavage(2 times per day).After 7 days of intragastric administration,aortic blood samples were extracted to collect serum samples using centrifugation,and serum samples from the same groups were combined to obtain the low-,medium-,and high-concentration Gushukang-containing and normal sera for the subsequent experiments.(1)RAW264.7 cells were cultured in six groups:normal serum was added to the control group;low,medium,and high concentration groups were added with low,medium,and high concentrations of Gushukang-containing serum,respectively;ML385,a nuclear factor erythroid 2-related factor 2(Nrf2)inhibitor was given in the Nrf2 inhibitor group;and t-BHQ,a Nrf2 activator,was added in the Nrf2 activator group.Cell viability was detected using the cell counting kit-8 assay.(2)The 3rd generation RAW 264.7 cells were cultured and divided into five groups:the blank control group was added with normal serum,the osteoclast group was added with receptor activator of nuclear factor κB ligand(RANKL),and the low-,medium-,and high-concentration groups were added with low-,medium-,and high-concentration Gushukang-containing serum based on the addition of RANKL.Tartrate-resistant acid phosphate staining was performed after 5 days of culture.(3)RAW264.7 cells were cultured and divided into five groups:blank control group was cultured with normal serum,osteoclast group cultured with normal serum and RANKL,high concentration+osteoclast group cultured with RANKL+high concentration Gushukang-containing serum,osteoclast+Nrf2 agonist group cultured with RANKL+t-BHQ,and high concentration+osteoclast+Nrf2 inhibitor group cultured with RANKL+high concentration Gushukang-containing serum+ML385.Western blot assay and determination of reactive oxygen content were performed after 5 days of culture. RESULTS AND CONCLUSION:The cell counting kit-8 results indicated that Gushukang-containing serum,NRF2 inhibitor or agonist had no significant effect on RAW264.7 cell viability.Tartrate-resistant acid phosphate staining results demonstrated that Gushukang-containing serum exhibited a concentration-dependent inhibitory effect on osteoclast differentiation.Western blot analysis and determination of reactive oxygen species revealed that compared with the blank control group,Nrf2 protein expression was decreased in the osteoclast group(P<0.05),while c-Fos and NFATc1 protein expression and reactive oxygen species content were elevated(P<0.05);compared with the osteoclast group,Nrf2 protein expression was elevated and reactive oxygen species content was decreased in the high-concentration+osteoclast group,osteoclast+Nrf2 agonist group,and high-concentration+osteoclast+Nrf2 inhibitor group(P<0.05),while c-Fos and NFATc1 protein expression was decreased in the high concentration+osteoclast group and osteoclast+Nrf2 agonist group(P<0.05);compared with the high concentration+osteoclast group,Nrf2 protein expression was decreased(P<0.05)and reactive oxygen species content was elevated(P<0.05)in the high concentration+osteoclast+Nrf2 inhibitor group.To conclude,Gushukang reduces reactive oxygen species production by activating Nrf2,thereby inhibiting downstream of the c-Fos/NFATc1 pathway and suppressing osteoclast differentiation.

Objective:To explore the effect of transcranial direct current stimulation (tDCS) on dysphagia in hemispheric stroke patients.Methods:Sixty-two hemispheric stroke patients with dysphagia were randomized into an ipsilateral group, a contralateral group and a bilateral group with 20 in each group. The ipsilateral and contralateral groups received tDCS over their ipsilesional and contralesional hemispheres, respectively, while in the bilateral group it was over both hemispheres. That was followed by conventional swallowing therapy. Before and after 2 weeks of the treatment, swallowing function was assessed using the modified Mann Assessment of Swallowing Ability (MMASA) and a Swallow Severity scale (SSS). Linear regressions were evaluated to highlight the factors most influencing recovery from post-stroke hemispheric dysphagia.Results:After the treatments, the average MMASA and SSS scores had increased significantly in all three groups. There was no significant difference in the average post-treatment MMASA and SSS scores between the ipsilateral and contralateral groups, but the bilateral group showed significantly better average post-treatment MMASA and SSS scores compared to the other two groups. Linear regression analysis confirmed that the tDCS protocol (group allocation) was a significant predictor of recovery.Conclusion:Bilateral tDCS can effectively promote the recovery of swallowing function after a hemispheric stroke. It demonstrates greater therapeutic benefits than unilateral tDCS.

Objective To explore the relationship between atherosclerosis(AS)and bone mineral density(BMD)as well as frac-ture risk in patients with type 2diabetes mellitus(T2DM).Methods A total of 380 T2DM patients aged 50-80 years were included and divided into AS group and non-AS group based on carotid intima-media thickness.Clinical indicators were compared between the two groups,and the relationship between AS and BMD as well as fracture risk was analyzed.Results The AS group had higher age,diabetes duration,incidence of osteopenia,10-year probability of major osteoporotic fracture(PMOF),and 10-year probability of hip fracture(PHF),while having lower femoral neck BMD than the non-AS group.In male T2DM patients,PMOF and PHF were positively correla-ted with age,diabetes duration,and AS lesions,and negatively correlated with total cholesterol,triglycerides,low-density lipoprotein,25-hydroxyvitamin D,type Ⅰ procollagen amino-terminal peptide,and BMD.In female T2DM patients,PMOF and PHF were posi-tively correlated with menopause duration,fasting insulin,and AS lesions,and negatively correlated with blood phosphorus and BMD.Regression analysis showed that osteopenia was an independent risk factor for AS lesions in male T2DM patients,AS lesions were an inde-pendent risk factor for BMD levels,and body mass index was an independent protective factor for BMD levels.In female T2DM patients,age,fasting insulin,and high-density lipoprotein were independent risk factors for BMD levels,and parathyroid hormone was an inde-pendent protective factor for BMD levels.Conclusion When T2DM patients have AS lesions,the incidence of low bone mass and osteo-porosis is higher,with decreased femoral neck BMD and increased PMOF and PHF.In male subjects,low bone mass and osteoporosis are independent risk factors for AS lesions,and AS lesions are also independent risk factors for BMD levels.

Objective To explore the relationship between atherosclerosis(AS)and bone mineral density(BMD)as well as frac-ture risk in patients with type 2diabetes mellitus(T2DM).Methods A total of 380 T2DM patients aged 50-80 years were included and divided into AS group and non-AS group based on carotid intima-media thickness.Clinical indicators were compared between the two groups,and the relationship between AS and BMD as well as fracture risk was analyzed.Results The AS group had higher age,diabetes duration,incidence of osteopenia,10-year probability of major osteoporotic fracture(PMOF),and 10-year probability of hip fracture(PHF),while having lower femoral neck BMD than the non-AS group.In male T2DM patients,PMOF and PHF were positively correla-ted with age,diabetes duration,and AS lesions,and negatively correlated with total cholesterol,triglycerides,low-density lipoprotein,25-hydroxyvitamin D,type Ⅰ procollagen amino-terminal peptide,and BMD.In female T2DM patients,PMOF and PHF were posi-tively correlated with menopause duration,fasting insulin,and AS lesions,and negatively correlated with blood phosphorus and BMD.Regression analysis showed that osteopenia was an independent risk factor for AS lesions in male T2DM patients,AS lesions were an inde-pendent risk factor for BMD levels,and body mass index was an independent protective factor for BMD levels.In female T2DM patients,age,fasting insulin,and high-density lipoprotein were independent risk factors for BMD levels,and parathyroid hormone was an inde-pendent protective factor for BMD levels.Conclusion When T2DM patients have AS lesions,the incidence of low bone mass and osteo-porosis is higher,with decreased femoral neck BMD and increased PMOF and PHF.In male subjects,low bone mass and osteoporosis are independent risk factors for AS lesions,and AS lesions are also independent risk factors for BMD levels.

Objective:To explore the effect of transcranial direct current stimulation (tDCS) on dysphagia in hemispheric stroke patients.Methods:Sixty-two hemispheric stroke patients with dysphagia were randomized into an ipsilateral group, a contralateral group and a bilateral group with 20 in each group. The ipsilateral and contralateral groups received tDCS over their ipsilesional and contralesional hemispheres, respectively, while in the bilateral group it was over both hemispheres. That was followed by conventional swallowing therapy. Before and after 2 weeks of the treatment, swallowing function was assessed using the modified Mann Assessment of Swallowing Ability (MMASA) and a Swallow Severity scale (SSS). Linear regressions were evaluated to highlight the factors most influencing recovery from post-stroke hemispheric dysphagia.Results:After the treatments, the average MMASA and SSS scores had increased significantly in all three groups. There was no significant difference in the average post-treatment MMASA and SSS scores between the ipsilateral and contralateral groups, but the bilateral group showed significantly better average post-treatment MMASA and SSS scores compared to the other two groups. Linear regression analysis confirmed that the tDCS protocol (group allocation) was a significant predictor of recovery.Conclusion:Bilateral tDCS can effectively promote the recovery of swallowing function after a hemispheric stroke. It demonstrates greater therapeutic benefits than unilateral tDCS.

Objective:To develop a double-set endotracheal intubation and evaluate its effectiveness.Methods:The development of a double-set endotracheal intubation was achieved by integrating endotracheal intubation with dental pads. From November 2023 to July 2024, convenience sampling was used to select 60 patients with endotracheal intubation admitted to the ICU of the First Hospital of Jiaxing as participants. Patients were divided into an experimental group ( n=30) and a control group ( n=30) using a random number table method. Experimental group was treated with a double-set endotracheal intubation, while control group was treated with a regular endotracheal intubation. The incidence of oral mucosal membrane pressure injury and oral ulcers during endotracheal intubation, the Chinese version of the modified Beck Oral Assessment score, plaque index, and endotracheal intubation duration were compared between two groups of ICU patients. Results:The duration of endotracheal intubation in experimental group was shorter than that in control group ( P<0.05), and the difference was statistically significant ( P<0.05). The number of cases with oral mucosal membrane pressure injury and oral ulcers in experimental group, as well as the Chinese version of the modified Beck Oral Assessment score, were lower than those in control group, and the differences were statistically significant ( P<0.05). There was no statistically significant difference in the plaque index between two groups ( P>0.05) . Conclusions:Double-set endotracheal intubation can effectively improve the efficiency of intubation in ICU patients, reduce the local pressure on oral tissues caused by intubation, improve the oral condition of patients, and have good clinical effects.

Objective:To develop a double-set endotracheal intubation and evaluate its effectiveness.Methods:The development of a double-set endotracheal intubation was achieved by integrating endotracheal intubation with dental pads. From November 2023 to July 2024, convenience sampling was used to select 60 patients with endotracheal intubation admitted to the ICU of the First Hospital of Jiaxing as participants. Patients were divided into an experimental group ( n=30) and a control group ( n=30) using a random number table method. Experimental group was treated with a double-set endotracheal intubation, while control group was treated with a regular endotracheal intubation. The incidence of oral mucosal membrane pressure injury and oral ulcers during endotracheal intubation, the Chinese version of the modified Beck Oral Assessment score, plaque index, and endotracheal intubation duration were compared between two groups of ICU patients. Results:The duration of endotracheal intubation in experimental group was shorter than that in control group ( P<0.05), and the difference was statistically significant ( P<0.05). The number of cases with oral mucosal membrane pressure injury and oral ulcers in experimental group, as well as the Chinese version of the modified Beck Oral Assessment score, were lower than those in control group, and the differences were statistically significant ( P<0.05). There was no statistically significant difference in the plaque index between two groups ( P>0.05) . Conclusions:Double-set endotracheal intubation can effectively improve the efficiency of intubation in ICU patients, reduce the local pressure on oral tissues caused by intubation, improve the oral condition of patients, and have good clinical effects.

Objective:To explore the anti-aging effects of berberine hydrochloride and underlying mechanism.Methods:Twelve 4-week-old C57BL6/J mice were divided into aging group (fed with normal chaw), aging+ BBR intervention group(fed with normal chaw containing 1 g/kg berberine hydrochloride). At the age of 64 weeks, all the experimental mice were executed. The liver tissues were made into paraffin sections for HE staining to observe the morphological and structural integrity of liver parenchymal cells for pathological evaluation. Immunofluorescence was used to detect p16 protein expression levels in liver. The expression levels of malondialdehyde(MDA), superoxide dismutase(SOD), and glutathione peroxidase(GSH-Px) were detected by colorimetry. The expression levels of interleukin(IL)-1β, IL-6, and tumor necrosis factor-α(TNF-α) in liver tissues and IL-8 in serum were detected by enzyme linked immunosorbent assay(ELISA) technique. The p16, p21, nuclear factor erythroid-2 related factor 2(Nrf2), nuclear factor-κB(NF-κB), phospho(p-)NF-κB, inhibitory κB(IκB)α, p-IκBα, and p38 mitogen-activated protein kinase(MAPK) protein expression levels in liver were detected by Western blotting. The same indices were tested in the 16-week-old mice as the young control group.Results:Compared with the young control group, the liver tissue in the aging control group exhibited morphological aging and antioxidant capacity were reduced( P<0.01), and inflammatory factors were increased( P<0.05 or P<0.01). Compared with the aging group, the liver tissues in the aging+ BBR intervention group were still maintain regular arrangement of hepatocytes, the p16 and p21 protein expression levels were significantly increased( P<0.05 or P<0.01), the antioxidant capacity were increased( P<0.05 or P<0.01), the level of inflammatory factors were decresed( P<0.05 or P<0.01), and the p38 MAPK/NF-κB pathway and Nrf2 pathway were inhibited( P<0.001). Conclusion:Berberine hydrochloride improves the aging morphology of the liver, potentially by suppressing the p38 MAPK/NF-κB pathway to reduce inflammation levels and inhibiting the Nrf2 pathway to ameliorate oxidative stress.

ObjectiveTo explore the amelioration of cognitive dysfunction in diabetes mellitus （DM） by Jianpi Qinghua prescription （JPQH） based on type 2 diabetes （T2DM） model rats. MethodFifty healthy male Wistar rats of SPF grade were randomly divided into control group （n=10） and experimental group （n=40）. The rats in the control group were fed conventionally， while those in the experimental group were fed on a high-sugar， high-fat diet for six weeks and administered with streptozotocin （STZ） for the induction of the DM model. The model rats were randomly divided into model group， sitagliptin group （1.2 g·L^-1）， pioglitazone group （0.8 g·L^-1）， and JPQH group （1.3 g·mL^-1）， with 10 rats in each group. After six weeks of drug intervention， the changes in body weight， blood glucose， and other related indexes of each group were recorded. Enzyme-linked immunosorbent assay （ELISA） was performed to detect the levels of tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， and interleukin-6 （IL-6） in the peripheral blood and brain. The Morris water maze test was used to evaluate the cognitive function in rats. Hematoxylin-eosin （HE） staining was used to observe the pathological morphology of the hippocampal CA region. The amyloid β-protein 40 （Aβ₄₀） level was detected by immunohistochemistry. The protein expression of t-tau and p-tau in hippocampal neurons of rats was detected by Western blot. ResultCompared with blank group， the body weight of model group was significantly decreased （P<0.05）， blood glucose level was significantly increased （P<0.01）， inflammatory cytokines TNF-α and IL-1β were increased （P<0.05）， learning and spatial ability were significantly decreased （P<0.01）， the arrangement of hippocampal cells was loose and disordered， and the intercellular space was significantly increased. The number of cells decreased significantly， and the expression of Aβ₄₀ increased significantly. and increased t-tau and p-tau protein content in the hippocampus （P<0.01）. Compared with model group， the JPQH group showed reduced blood glucose （P<0.01）， decreased TNF-α and IL-1β levels in the peripheral blood and cerebrospinal fluid （P<0.05）， a downward trend of IL-6 without a statistical difference， improved learning and spatial memory ability （P<0.01）， densely arranged cells in the hippocampal CA1 area， increased cell number， reduced Aβ₄₀ expression， and decreased p-tau protein expression （P<0.05）. ConclusionJPQH can prevent cognitive dysfunction in DM by reducing inflammatory factor levels， decreasing neurotoxicity caused by Aβ₄₀ deposition， and inhibiting hyperphosphorylation of tau protein in DM rats.