Objective To study the effect of stachydrine on cardiomyocyte damage induced by high glucose(HG)and its regulation of Hippo-Yes-associated protein(YAP)signaling pathway.Methods Rat myocardial cells H9C2 were stimulated by HG with 30 mmol/L glucose to establish the myocardial cell injury model,and then treated with 0.05,0.10,0.15,0.20 mmol/L of stachydrine and their activities were detected by CCK-8 method,and the action concentration of stachycarine was screened.Then H9C2 was grouped into control(ctrl)group,HG group,low concentration stachydrine group,high concentration stachydrine group,and high concentration stachydrine+Hippo-YAP signaling pathway inhibitor(TDI-011536)group.Except for the ctrl group cultured with 5 mmol/L glucose,the other 4 groups were cultured with 30 mmol/L glucose,and the low and high concentration stachydrine groups were cultured with 0.05 and 0.10 mmol/L threonine for 24h,respectively.The high concentration stachydrine+TDI-011536 group were cultured with 0.10 mmol/L stachydrine and 3.00 μmol/L TDI-011536 for 24h.CCK-8 method was applied to detect cell proliferation.Flow cytometry was applied to detect apoptosis.ELISA were applied to detect the level of malondialdehyde(MDA)and superoxide dismutase(SOD).Western blot was applied to detect the level of phosphorylated YAP(p-YAP),YAP,proliferating cell nuclear antigen(PCNA),and B-cell lymphoma-2(Bcl-2)proteins.Results Compared with ctrl group,the survival rate of H9C2 cells induced by HG was significantly increased by 0.05,0.10,0.15,0.20 mmol/L stachydrine treatment,and the differences were statistically significant(t=8.32～29.67,all P<0.01).The 50%inhibitory concentration(IC50)value of stachydrine was about 0.09 mmol/L,and the concentrations of 0.05 mmol/L and 0.10 mmol/L close to and lower than IC50 were selected as the concentrations of hydrostachyine for subsequent experiments.Compared with the ctrl group,the survival rate in HG group was significantly decreased(t=44.32,P<0.01).Compared with HG group,the cell survival rate of low concentration stachydrine group and high concentration stachydrine group was significantly increased(t=10.06,21.66,all P<0.01).Compared with the high concentration stachydrine group,the cell survival rate in the high-concentration stachydrine+TDI-011536 group was significantly decreased(t=9.54,P<0.01),and the differences were statistically significant,respectively.Compared with ctrl group,the apoptosis rate of HG group was significantly increased(t=36.74,P<0.01).Compared with HG group,the apoptosis rate of low concentration stachydrine group and high concentration stachydrine group was significantly decreased(t=11.04,26.78,all P<0.01).Compared with the high concentration threonine group,the apoptosis rate of the high concentration stachydrine+TDI-011536 group was significantly increased(t=9.96,P<0.01),and the differences were statistically significant,respectively.Compared with ctrl group,SOD level in HG group was significantly decreased,MDA levels were significantly increased(t=18.85,29.12,all P<0.01).Compared with HG group,SOD level were significantly increased in low concentration stachydrine groups and high concentration stachydrine groups(t=6.59,9.86,all P<0.01),MDA level were significantly decreased(t=13.45,23.36,all P<0.01).Compared with the high concentration stachydrine group,the SOD level in the high concentration hydrostatin+TDI-011536 group was significantly decreased.MDA levels were significantly increased,and the differences were statistically significant(t=5.30,6.98,all P<0.01),respectively.Compared with ctrl group,the level of p-YAP,p-YAP/YAP,PCNA,Bcl-2 protein were significantly decreased,and the level of YAP protein was significantly increased(t=15.36～45.00,all P<0.01).Compared with HG group,the level of p-YAP,p-YAP/YAP,PCNA,Bcl-2 protein were significantly increased in low concentration stachydrine group and high concentration stachydrine group,the level of YAP protein levels were significantly decreased(t=5.51～25.15,all P<0.01).Compared with the high concentration stachydrine group,the level of p-YAP,p-YAP/YAP,PCNA,Bcl-2 protein in the high concentration hydrothreonine+TDI-011536 group were significantly decreased,the level of YAP protein significantly increased,the differences were statistically significant(t=4.27～11.25,all P<0.05).Conclusion Stachydrine may inhibit oxidative stress and apoptosis by activating Hippo-YAP signaling pathway,thereby ameliorating HG-induced myocardial cell damage.

Objective:To explore the characteristics of gut microbiota changes in individuals with Sarcopenic Obesity(SO)based on 16S rRNA sequencing.Methods:This cross-sectional study was conducted in Chongming District, Shanghai from April to November 2021.Fecal samples were collected from 20 elderly SO patients (case group)and 40 elderly non-SO individuals(control group)for 16S rRNA sequencing in order to analyze the diversity, structural composition, and species differences of gut microbiota, and then to predict the differential metabolic functions of the gut microbiota.Results:A total of 60 subjects were included.The case group consisted of 20 individuals(15 males and 5 females)with an average age of 73.15 ± 4.09 years; the control group included 40 individuals(20 males and 20 females)with an average age of 71.20 ± 4.12 years.The α-diversity analysis revealed that the richness indices ACE and Chao 1 of the case group were significantly lower than those of the control group( P<0.05), while the diversity indices Shannon and Simpson showed a trend of being lower in the case group, but the differences were not statistically significant( P>0.05). Principal coordinate analysis based on the Unweighted-unifrac distance metric demonstrated a statistically significant difference in β-diversity between the two groups( P=0.003). The structure and composition of the gut microbiota in the case group were altered, with a significant reduction in the relative abundance of the Blautia genus in the case group( P<0.05). LEfSe analysis identified 5 and 16 enriched microbial species in the case and control groups, respectively (Linear Discriminant Analysis score >2, P<0.05). Additionally, PICRUSt2 functional prediction revealed significant differences( P<0.05)in metabolic pathways between the two groups, including quorum sensing, fat digestion and absorption, and folate biosynthesis. Conclusions:The gut microbiota in elderly SO patients is disordered, mainly manifested as a decrease in diversity and characteristic changes in structural composition, as well as a reduction in the abundance of the beneficial bacterium Blautia.The Progression of SO is closely associated with gut microbiota metabolic disturbances, and targeting the gut microbiota is expected to become a novel therapeutic approach for SO.

Objective:It analyzes the flow characteristics and spatial distribution patterns of intra-provincial cross-regional medical patients in Guangdong,while comparing hospitalization cost differences between local and non-local care.Methods:Based on 2022-2023 cross-regional medical data from Guangdong,spatial visualization and descriptive analysis were conducted combined with Mann-Whitney U test to compare the cost burden.Results:Cross-Regional patients in Guangdong primarily flow to Guangzhou,accounting for 77.8％of the total intra-provincial cross-regional hospitalizations.Cross-regional medical expenses were significantly higher than the insured locations,with notable differences in mean total hospitalization costs and personal out-of-pocket payment ratios.Conclusion:Refining pre-approval procedures for cross-regional care,enhancing specialized medical departments in under-resourced regions,and strengthening coordination between healthcare insurance systems and primary healthcare strengthening initiatives are needed to promote balanced medical resource allocation and reduce the cost burden on patients.

OBJECTIVE To investigate the ameliorative effect of Pinelliae Rhizoma Praeparatum Cum Alumine and its effective components on inflammatory response in mice with acute lung injury(ALI).METHODS A mice model of acute lung injury induced by lipopolysaccharide(LPS)was used as the in vivo animal model.The levels of inflammatory factors TNF-α,IL-6,and IL-1β,the total protein concentration,the number of inflammatory cells in bronchoalveolar lavage fluid,the ratio of lung wet weight to dry weight,and the pathological damage of lung tissues were taken as the evaluation indexes.RAW264.7 macrophages induced by LPS were used as the in vitro experimental model,with the mRNA levels of intracellular inflammatory factors TNF-α,IL-1β,and IL-6 as indicators,systematic solvent extraction and silica gel column chromatography were used to separate and screen the effective parts and components of Pinelliae Rhizoma Praeparatum Cum Alumine for treating acute lung injury.The component composition of the effective fractions was analyzed by mass spectrometry,the content of cyclic dipeptide components in the effective fraction was determined,and the anti-in-flammatory activity of cyclic dipeptides was verified in vitro.RESULTS The ethyl acetate extraction fraction obtained by extracting the supernatant of water extraction and ethanol precipitation of Pinelliae Rhizoma Praeparatum Cum Alumine(referred to as the ethyl acetate fraction for short)significantly improved the inflammatory response in mice with acute lung injury and significantly reduced the mRNA levels of TNF-α,IL-1β,and IL-6 in RAW264.7 cells induced by LPS.The Fr.1 fraction isolated from the ethyl acetate frac-tion by silica gel column chromatography had anti-inflammatory effects in vitro.Mass spectrometry determined the content of two main cyclic dipeptide components,cyclo-(Pro-Phe)and cyclo-(Pro-Val),in the Fr.1 fraction.After mixing these two cyclic dipeptide components in the molar ratio in the Fr.1 fraction and administering them in vitro,the results showed that the combination of these two cyclic dipeptide components could significantly inhibit the mRNA expression levels of TNF-α,IL-1β,and IL-6 in RAW264.7 cells induced by LPS.CONCLUSION Cyclic dipeptide component is one of the main material basis categories for the anti-inflammatory effect of Pinelliae Rhizoma Praeparatum Cum Alumine.

Objective:To evaluate the efficacy and safety of remedial pedicled rectus abdominis myocutaneous flap transplantation for breast reconstruction.Methods:This retrospective study included 23 female breast cancer patients who underwent autologous breast reconstruction with a remedial pedicled rectus abdominis myocutaneous flap at Hunan Cancer Hospital from June 2022 to June 2023. The patients' ages ranged from 24 to 56 years, with a mean age of (43.2±1.3) years. All patients had previously undergone mastectomy and tumor resection before receiving the remedial pedicled rectus abdominis myocutaneous flap reconstruction. Follow-up was conducted through outpatient visits and telephone interviews, with a median follow-up period of 10 (6-15) months. Observation indicators included survival rate of flaps, patient satisfaction rate and aesthetic outcomes. Adverse reactions including but not limited to flap contracture and deformation, during and following the surgical procedure were also recorded.Results:The remedial pedicled rectus abdominis myocutaneous flap technique was utilized for autologous breast reconstruction in 23 patients. Specifically, this approach was chosen due to two primary reasons: the perforator distance from the vascular pedicle being excessively long in 18 cases, and suboptimal characteristics of the main pedicle in 5 cases. Two types of flap configurations were employed. For 15 patients, a unilateral remedial pedicled rectus abdominis myocutaneous flap was used, with an average flap size of (21.5±0.8) cm in length and (11.2±1.3) cm in width. All flaps demonstrated adequate blood supply without the need for additional vascular anastomosis. For the remaining 8 patients, a combination of a unilateral remedial pedicled rectus abdominis myocutaneous flap and a contralateral free lower abdominal flap was utilized, resulting in an average flap size of (29.3±0.7) cm in length and (12.0±1.8) cm in width. In these cases, the inferior epigastric vessels of the free lower abdominal flap were successfully anastomosed to the internal thoracic vessels of the recipient site, ensuring all flaps survived postoperatively. The survival rate of flaps in 23 patients was 100% (23/23), and the patient satisfaction rate was 100% (23/23). Postoperative evaluation revealed that the reconstructed breasts exhibited satisfactory shape and texture, with no instances of flap contracture or deformation. The reconstructed breasts were largely symmetrical with the contralateral side, leaving only linear scars at the donor and recipient sites. Importantly, there was no significant impact on abdominal wall function, and no adverse reactions such as flap contracture or impaired abdominal wall function were observed in any of the patients.Conclusion:When the characteristics of the perforating vessels in the lower abdomen are suboptimal, the use of a remedial pedicled rectus abdominis myocutaneous flap technique for breast reconstruction can ensure satisfactory outcomes and significantly enhance surgical safety.

Objective To investigate the distribution and antimicrobial resistance profiles of clinically isolated Haemophilus influenzae and Moraxella catarrhalis in hospitals across China from 2015 to 2021,and provide evidence for rational use of antimicrobial agents.Methods Data of H.influenzae and M.catarrhalis strains isolated from 2015 to 2021 in CHINET program were collected for analysis,and antimicrobial susceptibility testing was performed by disc diffusion method or automated systems according to the uniform protocol of CHINET.The results were interpreted according to the CLSI breakpoints in 2022.Beta-lactamases was detected by using nitrocefin disk.Results From 2015 to 2021,a total of 43 642 strains of Haemophilus species were isolated,accounting for 2.91％of the total clinical isolates and 4.07％of Gram-negative bacteria in CHINET program.Among the 40 437 strains of H.influenzae,66.89％were isolated from children and 33.11％were isolated from adults.More than 90％of the H.influenzae strains were isolated from respiratory tract specimens.The prevalence of β-lactamase was 53.79％in H.influenzae strains.The H.influenzae strains isolated from children showed higher resistance rate than the strains isolated from adults.Overall,779 strains of H.influenzae did not produce β-lactamase but were resistant to ampicillin(BLNAR).Beta-lactamase-producing strains showed significantly higher resistance rates to these antimicrobial agents than the β-lactamase-nonproducing strains.Of the 16 191 M.catarrhalis strains,80.06％were isolated from children and 19.94％isolated from adults.M.catarrhalis strains were mostly susceptible to both amoxicillin-clavulanic acid and cefuroxime,evidenced by resistance rate lower than 2.0％.Conclusions The emergence of antibiotic-resistant H.influenzae due to β-lactamase production poses a challenge for clinical anti-infective treatment.Therefore,it is very important to implement antibiotic resistance surveillance for H.influenzae and guide rational antibiotic use.All local clinical microbiology laboratories should actively improve antibiotic susceptibility testing and strengthen antibiotic resistance surveillance for H.influenzae.

BACKGROUND:In the occurrence and development of demyelinating diseases of the central nervous system,neuroinflammation caused by microglia is the main pathological feature,so inhibiting the inflammatory response is very important to alleviate demyelination.Hydroxysafflor yellow A can protect the blood-brain barrier,inhibit neuronal apoptosis,and improve neurological function.OBJECTIVE:To explore the mechanism of hydroxysafflor yellow A inhibiting bicyclohexanone oxalyl dihydrazone-induced demyelination in mice.METHODS:(1)In vivo:Thirty healthy male C57BL/6 mice were randomly divided into three groups:normal group,cuprizone group,and hydroxysafflor yellow A group.The mice in the cuprizone group and the hydroxysafflor yellow A group were fed with 0.2％cuprizone diet for 6 weeks to establish mouse models of demyelination.The mice in the normal group were fed with normal diet.At the end of the 4th week,the mice in the hydroxysafflor yellow A group were intraperitoneally injected with hydroxysafflor yellow A 20 mg/kg per day.The mice in the normal and cuprizone groups were intraperitoneally injected with normal saline for 2 weeks.The behavioral changes of mice were evaluated by open field test and elevated plus maze test.The loss of myelin sheath in corpus callosum was detected by black gold staining,myelin basic protein and degraded myelin basic protein immunofluorescence staining.The activation of microglia and the expression of inflammatory factors were detected by I ba-1 immunofluorescence staining and ELISA,respectively.The protein expression levels of Toll-like receptor 4,myeloid differentiation factor 88,and nuclear factor κB p65 in the brain of mice in each group were detected by western blot assay.(2)In vitro experiment:The inflammation model of BV2 microglia was established by lipopolysaccharide induction.BV2 cells were divided into normal group,lipopolysaccharide group(1 μg/mL),and lipopolysaccharide(1 μg/mL)+hydroxysafflor yellow A(25 μmol/L)group.The expression levels of tumor necrosis factor α and interleukin 6 in the supernatant were detected by ELISA.RESULTS AND CONCLUSION:(1)Compared with the normal group,the mice in the cuprizone group had severe anxiety,abnormal autonomic movement ability,and a large amount of myelin sheath loss in the corpus callosum.The average fluorescence intensity of myelin basic protein was significantly reduced,and the average fluorescence intensity of degraded myelin basic protein was significantly increased.The number of lba1+microglia increased,the contents of interleukin 1β,tumor necrosis factor α,and interleukin 6 in the brain increased,and the protein expression levels of Toll-like receptor 4,myeloid differentiation factor 88,and nuclear factor κB p65 increased significantly.The above symptoms and indexes of mice were reversed after hydroxysafflor yellow A treatment.(2)Hydroxysafflor yellow A significantly inhibited the expression of inflammatory factors such as tumor necrosis factor α,and interleukin 6 induced by lipopolysaccharide in BV2 microglia.(3)The above results demonstrate that hydroxysafflor yellow A can significantly improve cuprizone-induced demyelination in mice.The mechanism of action is related to the inhibition of microglial activation-mediated inflammatory response through the Toll-like receptor 4/myeloid differentiation factor 88/nuclear factor κB p65 signaling pathway.

Objective To investigate the distribution and antimicrobial resistance profiles of clinically isolated Haemophilus influenzae and Moraxella catarrhalis in hospitals across China from 2015 to 2021,and provide evidence for rational use of antimicrobial agents.Methods Data of H.influenzae and M.catarrhalis strains isolated from 2015 to 2021 in CHINET program were collected for analysis,and antimicrobial susceptibility testing was performed by disc diffusion method or automated systems according to the uniform protocol of CHINET.The results were interpreted according to the CLSI breakpoints in 2022.Beta-lactamases was detected by using nitrocefin disk.Results From 2015 to 2021,a total of 43 642 strains of Haemophilus species were isolated,accounting for 2.91％of the total clinical isolates and 4.07％of Gram-negative bacteria in CHINET program.Among the 40 437 strains of H.influenzae,66.89％were isolated from children and 33.11％were isolated from adults.More than 90％of the H.influenzae strains were isolated from respiratory tract specimens.The prevalence of β-lactamase was 53.79％in H.influenzae strains.The H.influenzae strains isolated from children showed higher resistance rate than the strains isolated from adults.Overall,779 strains of H.influenzae did not produce β-lactamase but were resistant to ampicillin(BLNAR).Beta-lactamase-producing strains showed significantly higher resistance rates to these antimicrobial agents than the β-lactamase-nonproducing strains.Of the 16 191 M.catarrhalis strains,80.06％were isolated from children and 19.94％isolated from adults.M.catarrhalis strains were mostly susceptible to both amoxicillin-clavulanic acid and cefuroxime,evidenced by resistance rate lower than 2.0％.Conclusions The emergence of antibiotic-resistant H.influenzae due to β-lactamase production poses a challenge for clinical anti-infective treatment.Therefore,it is very important to implement antibiotic resistance surveillance for H.influenzae and guide rational antibiotic use.All local clinical microbiology laboratories should actively improve antibiotic susceptibility testing and strengthen antibiotic resistance surveillance for H.influenzae.

Objective:It analyzes the flow characteristics and spatial distribution patterns of intra-provincial cross-regional medical patients in Guangdong,while comparing hospitalization cost differences between local and non-local care.Methods:Based on 2022-2023 cross-regional medical data from Guangdong,spatial visualization and descriptive analysis were conducted combined with Mann-Whitney U test to compare the cost burden.Results:Cross-Regional patients in Guangdong primarily flow to Guangzhou,accounting for 77.8％of the total intra-provincial cross-regional hospitalizations.Cross-regional medical expenses were significantly higher than the insured locations,with notable differences in mean total hospitalization costs and personal out-of-pocket payment ratios.Conclusion:Refining pre-approval procedures for cross-regional care,enhancing specialized medical departments in under-resourced regions,and strengthening coordination between healthcare insurance systems and primary healthcare strengthening initiatives are needed to promote balanced medical resource allocation and reduce the cost burden on patients.

Objective To study the effect of stachydrine on cardiomyocyte damage induced by high glucose(HG)and its regulation of Hippo-Yes-associated protein(YAP)signaling pathway.Methods Rat myocardial cells H9C2 were stimulated by HG with 30 mmol/L glucose to establish the myocardial cell injury model,and then treated with 0.05,0.10,0.15,0.20 mmol/L of stachydrine and their activities were detected by CCK-8 method,and the action concentration of stachycarine was screened.Then H9C2 was grouped into control(ctrl)group,HG group,low concentration stachydrine group,high concentration stachydrine group,and high concentration stachydrine+Hippo-YAP signaling pathway inhibitor(TDI-011536)group.Except for the ctrl group cultured with 5 mmol/L glucose,the other 4 groups were cultured with 30 mmol/L glucose,and the low and high concentration stachydrine groups were cultured with 0.05 and 0.10 mmol/L threonine for 24h,respectively.The high concentration stachydrine+TDI-011536 group were cultured with 0.10 mmol/L stachydrine and 3.00 μmol/L TDI-011536 for 24h.CCK-8 method was applied to detect cell proliferation.Flow cytometry was applied to detect apoptosis.ELISA were applied to detect the level of malondialdehyde(MDA)and superoxide dismutase(SOD).Western blot was applied to detect the level of phosphorylated YAP(p-YAP),YAP,proliferating cell nuclear antigen(PCNA),and B-cell lymphoma-2(Bcl-2)proteins.Results Compared with ctrl group,the survival rate of H9C2 cells induced by HG was significantly increased by 0.05,0.10,0.15,0.20 mmol/L stachydrine treatment,and the differences were statistically significant(t=8.32～29.67,all P<0.01).The 50%inhibitory concentration(IC50)value of stachydrine was about 0.09 mmol/L,and the concentrations of 0.05 mmol/L and 0.10 mmol/L close to and lower than IC50 were selected as the concentrations of hydrostachyine for subsequent experiments.Compared with the ctrl group,the survival rate in HG group was significantly decreased(t=44.32,P<0.01).Compared with HG group,the cell survival rate of low concentration stachydrine group and high concentration stachydrine group was significantly increased(t=10.06,21.66,all P<0.01).Compared with the high concentration stachydrine group,the cell survival rate in the high-concentration stachydrine+TDI-011536 group was significantly decreased(t=9.54,P<0.01),and the differences were statistically significant,respectively.Compared with ctrl group,the apoptosis rate of HG group was significantly increased(t=36.74,P<0.01).Compared with HG group,the apoptosis rate of low concentration stachydrine group and high concentration stachydrine group was significantly decreased(t=11.04,26.78,all P<0.01).Compared with the high concentration threonine group,the apoptosis rate of the high concentration stachydrine+TDI-011536 group was significantly increased(t=9.96,P<0.01),and the differences were statistically significant,respectively.Compared with ctrl group,SOD level in HG group was significantly decreased,MDA levels were significantly increased(t=18.85,29.12,all P<0.01).Compared with HG group,SOD level were significantly increased in low concentration stachydrine groups and high concentration stachydrine groups(t=6.59,9.86,all P<0.01),MDA level were significantly decreased(t=13.45,23.36,all P<0.01).Compared with the high concentration stachydrine group,the SOD level in the high concentration hydrostatin+TDI-011536 group was significantly decreased.MDA levels were significantly increased,and the differences were statistically significant(t=5.30,6.98,all P<0.01),respectively.Compared with ctrl group,the level of p-YAP,p-YAP/YAP,PCNA,Bcl-2 protein were significantly decreased,and the level of YAP protein was significantly increased(t=15.36～45.00,all P<0.01).Compared with HG group,the level of p-YAP,p-YAP/YAP,PCNA,Bcl-2 protein were significantly increased in low concentration stachydrine group and high concentration stachydrine group,the level of YAP protein levels were significantly decreased(t=5.51～25.15,all P<0.01).Compared with the high concentration stachydrine group,the level of p-YAP,p-YAP/YAP,PCNA,Bcl-2 protein in the high concentration hydrothreonine+TDI-011536 group were significantly decreased,the level of YAP protein significantly increased,the differences were statistically significant(t=4.27～11.25,all P<0.05).Conclusion Stachydrine may inhibit oxidative stress and apoptosis by activating Hippo-YAP signaling pathway,thereby ameliorating HG-induced myocardial cell damage.