ObjectiveTo explore the potential mechanism of Xianglian Huazhuo prescription （XLHZ） in treating chronic atrophic gastritis （CAG） by regulating cell cycle and inhibiting proliferation， using bioinformatics technology and animal experiments. MethodsDifferential expressed genes （DEGs） related to CAG were screened using GEO database and GEO2R tool. Weighted gene co-expression network analysis （WGCNA） was employed to search for hub genes of CAG. These hub genes were intersected with cell cycle proliferation based on GeneCards database. Eenrichment analysis of the intersecting genes was performed to obtain signaling pathways and biological processes related to CAG. Protein protein interaction （PPI） analysis of genes was conducted using the Protein Interaction Platform （STRING） database to search the super hub gene （hub 2.0）， and animal experiments were conducted for further validation. Fourteen of 70 male Wistar rats were randomly selected as the normal group， and the remaining 56 rats were prepared by the combined modeling method of "starvation disorder+N-methyl-N-nitro-N-nitrosoguanidine （MNNG） + sodium salicylate". The successfully modeled rats were randomly divided into the model group， XLHZ-H， XLHZ-M， and XLHZ-L groups （36， 18， 9 g·kg^-1， respectively）， and Morodan group （1.4 g·kg^-1）. Each group was given corresponding intervention for 60 days. Hematoxylin-eosin （HE） staining was used to observe the histopathological changes of gastric mucosa in rats. The ultrastructure of gastric mucosal tissue cells was observed by transmission electron microscopy. The relative expression levels of TGF-β₁， Smad2 and Smad3 proteins， S/G₂/M phase marker geminin and proliferation marker MCM2 were detected by Western blot in gastric mucosal tissue， and Spearman correlation analysis was performed. ResultsA total of 15 hub 2.0 genes were identified， including TGF-β₁， suggesting the involvement of the TGF-β₁ signaling pathway in the CAG pathogenesis. Compared with the normal group， the expressions of TGF-β₁， Smad2， geminin and MCM2 proteins in the gastric mucosa tissue of the model group were increased （P<0.05）， and the expression of Smad3 protein was decreased （P<0.05）. Compared with the model group， the expressions of TGF-β₁ and geminin in the gastric mucosa were decreased in the drug groups （P<0.05）. The XLHZ-M group， XLHZ-H group and Morodan group had significantly decreased protein expression of Smad2 and MCM2 （P<0.05）. The protein expression of Smad3 was significantly increased in XLHZ-M， XLHZ-H， and Morodan groups （P<0.05）. Spearman correlation analysis showed that Smad3 was negatively correlated with other indicators， and positively correlated with other indicators （P<0.01）. ConclusionXLHZ may inhibit TGF-β₁/Smads signaling pathway， regulate cell cycle， and inhibit proliferation in the treatment of CAG.

ObjectiveTo explore the potential mechanism of Xianglian Huazhuo prescription （XLHZ） in treating chronic atrophic gastritis （CAG） by regulating cell cycle and inhibiting proliferation， using bioinformatics technology and animal experiments. MethodsDifferential expressed genes （DEGs） related to CAG were screened using GEO database and GEO2R tool. Weighted gene co-expression network analysis （WGCNA） was employed to search for hub genes of CAG. These hub genes were intersected with cell cycle proliferation based on GeneCards database. Eenrichment analysis of the intersecting genes was performed to obtain signaling pathways and biological processes related to CAG. Protein protein interaction （PPI） analysis of genes was conducted using the Protein Interaction Platform （STRING） database to search the super hub gene （hub 2.0）， and animal experiments were conducted for further validation. Fourteen of 70 male Wistar rats were randomly selected as the normal group， and the remaining 56 rats were prepared by the combined modeling method of "starvation disorder+N-methyl-N-nitro-N-nitrosoguanidine （MNNG） + sodium salicylate". The successfully modeled rats were randomly divided into the model group， XLHZ-H， XLHZ-M， and XLHZ-L groups （36， 18， 9 g·kg^-1， respectively）， and Morodan group （1.4 g·kg^-1）. Each group was given corresponding intervention for 60 days. Hematoxylin-eosin （HE） staining was used to observe the histopathological changes of gastric mucosa in rats. The ultrastructure of gastric mucosal tissue cells was observed by transmission electron microscopy. The relative expression levels of TGF-β₁， Smad2 and Smad3 proteins， S/G₂/M phase marker geminin and proliferation marker MCM2 were detected by Western blot in gastric mucosal tissue， and Spearman correlation analysis was performed. ResultsA total of 15 hub 2.0 genes were identified， including TGF-β₁， suggesting the involvement of the TGF-β₁ signaling pathway in the CAG pathogenesis. Compared with the normal group， the expressions of TGF-β₁， Smad2， geminin and MCM2 proteins in the gastric mucosa tissue of the model group were increased （P<0.05）， and the expression of Smad3 protein was decreased （P<0.05）. Compared with the model group， the expressions of TGF-β₁ and geminin in the gastric mucosa were decreased in the drug groups （P<0.05）. The XLHZ-M group， XLHZ-H group and Morodan group had significantly decreased protein expression of Smad2 and MCM2 （P<0.05）. The protein expression of Smad3 was significantly increased in XLHZ-M， XLHZ-H， and Morodan groups （P<0.05）. Spearman correlation analysis showed that Smad3 was negatively correlated with other indicators， and positively correlated with other indicators （P<0.01）. ConclusionXLHZ may inhibit TGF-β₁/Smads signaling pathway， regulate cell cycle， and inhibit proliferation in the treatment of CAG.

Objective To investigate the therapeutic mechanism of Dingkun Dan(DKD)in polycystic ovary syndrome(PCOS)by examining the effects of microRNA-30d-5p on ovarian granulosa cells(GCs).Methods A PCOS rat model was established using dehydroepiandrosterone(DHEA).Normal rat GCs and PCOS rat GCs were cultured in vitro and divided into four groups:blank control group,model group,low-dose DKD group,and high-dose DKD group.After grouping,GCs viability was assessed using the methyl thiazolyl tetrazolium(MTT)assay,GCs apoptosis was analyzed by flow cytometry,and the gene expression of transforming growth factor β1(TGF-β1),Smad2,Smad3,and microRNA-30d-5p in GCs was measured by real-time polymerase chain reaction(RT-PCR),protein expression of TGF-β1,Smad2,and Smad3 in GCs was detected by Western Blot.Results Compared with the blank control group,the model group exhibited significantly decreased GCs viability,increased GCs apoptosis,upregulated mRNA and protein expression of TGF-β1,Smad2,and Smad3,and downregulated microRNA-30d-5p expression,the differences were statistically significant(P＜0.01).Compared with the model group,both low-and high-dose DKD groups showed increased GCs viability,reduced GCs apoptosis,downregulated mRNA and protein levels of TGF-β1 Smad2 and Smad3,elevated microRNA-30d-5p expression,and the differences were statistically significant(P＜0.05 or P＜0.01).Conclusion DKD promotes GCs proliferation by targeting Smad2 via microRNA-30d-5p,suggesting a potential therapeutic role in PCOS-related ovulatory dysfunction.

BACKGROUND: Asthma is a common chronic inflammatory airway disease and intermittent hypoxia is increasingly recognized as a factor that may impact disease progression. The present study investigated whether intermittent hypoxia (IH) could aggravate asthma by promoting hypoxia-inducible factor-1α (HIF-1α)/nucleotide-binding oligomerization domain (NOD)-like receptor pyrin domain-containing protein 3 (NLRP3)/interleukin (IL)-1β-dependent pyroptosis and the inflammatory response and further elucidated the underlying molecular mechanisms involved. METHODS: A total of 49 patients diagnosed with severe bronchial asthma and diagnosed by polysomnography were enrolled at Tongji Hospital, Tongji Medical College of Huazhong University of Science and Technology, between January 2022 and December 2022, and their general data and induced sputum were collected. BEAS-2B cells were treated with IL-13 and subjected to IH. An ovalbumin (OVA)-treated mouse model was also used to assess the effects of chronic intermittent hypoxia (CIH) on asthma. Pyroptosis, the inflammatory response, and related signalling pathways were assessed in vivo and in vitro . RESULTS: In this study, as the apnoea and hypopnea index (AHI) increased, the proportion of patients with uncontrolled asthma increased. The proportions of neutrophils and the levels of IL-6, IL-8, HIF-1α and NLRP3 in induced sputum were related to the AHI. NLRP3-mediated pyroptosis, which could be mediated by the HIF-1α signalling pathway, was activated in IL-13 plus IH-treated BEAS-2B cells and in the lungs of OVA/CIH mice. HIF-1α downregulation significantly reduced lung pyroptosis and ameliorated neutrophil inflammation by modulating the NLRP3/IL-1β pathway both in vitro and in vivo . Similarly, pretreatment with LW6, an inhibitor of HIF-1α, effectively blocked the generation of inflammatory cytokines in neutrophils. In addition, administration of the NLRP3 activator nigericin obviously increased lung neutrophil inflammation. CONCLUSIONS Obstructive sleep apnoea-hypopnea syndrome (OSAHS) is a risk factor for asthma exacerbation. IH aggravates neutrophil inflammation in asthma via NLRP3/IL-1β-dependent pyroptosis mediated by the HIF-1α signalling pathway, which should be considered a potential therapeutic target for the treatment of asthma with OSAHS.
NLR Family, Pyrin Domain-Containing 3 Protein/metabolism* ; Humans ; Asthma/metabolism* ; Animals ; Pyroptosis/physiology* ; Hypoxia-Inducible Factor 1, alpha Subunit/metabolism* ; Mice ; Signal Transduction/physiology* ; Male ; Hypoxia/metabolism* ; Female ; Interleukin-1beta/metabolism* ; Adult ; Inflammation/metabolism* ; Middle Aged ; Mice, Inbred C57BL

Objective To assess the effect of platycodin D(PD)for alleviating pulmonary fibrosis in mice and explore the underlying mechanism.Methods C57BL/6J mouse models of pulmonary fibrosis induced by bleomycin injection into the airway were treated with daily intragastric administration of 10 mg/kg PD for 28 days.The changes of pulmonary fibrosis and the expression and distribution of transient receptor potential cation channel subfamily C member 6(TRPC6)were evaluated with immunohistochemistry,HE staining and Sirius Red staining.Western blotting was used to detect α-SMA expression in the lung tissues of the mice.Primary cultures of mouse lung fibroblasts were pretreated with PD(2.5,5.0,and 10 μmol/L)or larixyl acetate(LA;10 μmol/L)before exposure to 10 ng/mL transforming growth factor-β1(TGF-β1),and the changes in cell survival rate,expressions of collagen I,α-SMA and TRPC6,reactive oxygen species(ROS)production,mitochondrial membrane potential,and cell proliferation capacity were assessed.Network pharmacology analysis was performed to explore the mechanism by which PD alleviated pulmonary fibrosis.Results PD treatment significantly alleviated pulmonary fibrosis and reduced α-SMA expression in BLM-induced mouse models(P<0.05).In TGF-β1-induced primary mouse lung fibroblasts,PD effectively inhibited the cell proliferation,reduced ROS production(P<0.0001),rescued the reduction of mitochondrial membrane potential(P<0.001),and inhibited the expressions of α-SMA and collagenⅠ(P<0.05).Network pharmacology analysis suggested that TRPC6 mediated the effect of PD for alleviating pulmonary fibrosis.Immunohistochemistry showed that PD significantly reduced TRPC6 expression in the lung tissues of BLM-induced mice.In primary mouse lung fibroblasts,PD significantly inhibited TGF-β1-induced TRPC6 expression(P<0.05),and LA treatment obviously lowered the expression levels of TRPC6,α-SMA and collagenⅠ(P<0.05).Conclusion PD alleviated pulmonary fibrosis in mice possibly by down-regulating TRPC6 and reducing ROS production.

Objective:To investigate the clinical, epidemic and laboratory characteristics of brucellosis in Hainan, and to summarize the experience of diagnosis and treatment.Methods:The epidemiological characteristics, clinical manifestations, laboratory results, treatment and prognosis of 27 inpatients with brucellosis admitted to the Second Affiliated Hospital of Hainan Medical University from January 2019 to December 2023 were retrospectively analyzed.Results:In 27 cases, the male to female ratio was 2.38 ∶ 1.00 (19 ∶ 8). Occupational distribution was dominated by farmers and herdsmen, with a total of 21 cases (77.8%). Twenty-one patients (77.8%) had a clear history of contact with cattle, sheep, and related products, and there was a phenomenon of family clustering. Eighteen patients (66.7%) developed symptoms from March to July. The clinical manifestations of the patients were lack of specificity, mainly fever (24 cases, 88.9%), dizziness and headache (13 cases, 48.1%), and low back pain (11 cases, 40.7%). Some patients also had symptoms and signs such as fatigue (8 cases), muscle pain (6 cases), chest tightness (4 cases), and splenomegaly (6 cases). Complications commonly included pulmonary infection (13 cases, 48.1%) and spinal involvement (11 cases, 40.7%). The positive rates of Brucella blood culture, tube agglutination test, Rose Bengal plate agglutination test, and cerebrospinal fluid or joint cavity effusion culture were 76.9% (20/26), 100.0% (21/21), 100.0% (11/11), and 3/4, respectively. The combination therapy of doxycycline and rifampicin showed good therapeutic effects, and 22 patients were cured. Conclusions:Hainan is not an epidemic area of brucellosis, and the clinical manifestations of the disease are not specific. The epidemic and clinical characteristics of the disease should be paid attention to in order to improve the level of diagnosis and treatment and control effectiveness.

Objective:To study the epidemiological and clinical characteristics of patients infected with Streptococcus suis in Hainan Province. Methods:Clinical data of patients diagnosed with human infection with Streptococcus suis at the Second Affiliated Hospital of Hainan Medical University from January 2017 to December 2023 were collected, and their epidemiological characteristics, clinical manifestations, auxiliary examination results, treatment and prognosis were retrospectively analyzed. Results:A total of 29 patients infected with Streptococcus suis were included, with 26 males (89.66%) and 3 females (10.34%). The age of the patients was (54.52 ± 10.89) years old, with the highest number of patients aged 51 to 60 years, accounting for 41.38% (12 cases). Farmers were the main occupation (72.41%, 21/29). Forty-four point eighty-three percent (13/29) of the patients had a history of direct or indirect contact with dead or sick pigs. The number of cases from January to December was 0, 0, 2, 2, 4, 8, 2, 3, 2, 2, 3, and 1, respectively. June was the peak period of incidence (27.59%, 8/29). The main clinical manifestations were fever (100.00%, 29/29), altered state of consciousness (51.72%, 15/29), headache (48.28%, 14/29), and hearing impairment (44.83%, 13/29). In clinical classification, common type, meningitis type, and mixed type accounted for 20.69% (6/29), 75.86% (22/29), and 3.45% (1/29), respectively. In the pathogen examination, all 29 patients underwent blood culture, and 18 patients (62.07%, 18/29) had positive blood culture; 26 patients underwent cerebrospinal fluid culture, and 8 cases (30.77%) tested positive for cerebrospinal fluid culture. Eighty-nine point sixty-six percent (26/29) of the patients were treated with ceftriaxone sodium; 55.17% (16/29) of the patients were treated with combined treatment of ceftriaxone sodium and glucocorticoids, including 11 cases treated with dexamethasone and 5 cases treated with methylprednisolone. After treatment, 7 patients were cured and discharged, and 22 patients showed improvement in their condition and discharged from hospital. At discharge, both blood and/or cerebrospinal fluid cultures turned negative. Telephone follow-up was conducted 6 months after discharge, and 20 patients had no sequelae, 9 patients still had hearing impairment, including one who also developed walking difficulties. Conclusions:The patients infected with Streptococcus suis in Hainan Province are mainly middle-aged, elderly, male and farmers, with June being the peak period of the disease. The main clinical manifestations are fever, altered state of consciousness, headache, and hearing impairment. After follow-up, there are still patients with residual hearing impairment. In clinical practice, patients with fever and hearing impairment should be considered as this disease as early as possible.

Objective:To investigate the clinical, epidemic and laboratory characteristics of brucellosis in Hainan, and to summarize the experience of diagnosis and treatment.Methods:The epidemiological characteristics, clinical manifestations, laboratory results, treatment and prognosis of 27 inpatients with brucellosis admitted to the Second Affiliated Hospital of Hainan Medical University from January 2019 to December 2023 were retrospectively analyzed.Results:In 27 cases, the male to female ratio was 2.38 ∶ 1.00 (19 ∶ 8). Occupational distribution was dominated by farmers and herdsmen, with a total of 21 cases (77.8%). Twenty-one patients (77.8%) had a clear history of contact with cattle, sheep, and related products, and there was a phenomenon of family clustering. Eighteen patients (66.7%) developed symptoms from March to July. The clinical manifestations of the patients were lack of specificity, mainly fever (24 cases, 88.9%), dizziness and headache (13 cases, 48.1%), and low back pain (11 cases, 40.7%). Some patients also had symptoms and signs such as fatigue (8 cases), muscle pain (6 cases), chest tightness (4 cases), and splenomegaly (6 cases). Complications commonly included pulmonary infection (13 cases, 48.1%) and spinal involvement (11 cases, 40.7%). The positive rates of Brucella blood culture, tube agglutination test, Rose Bengal plate agglutination test, and cerebrospinal fluid or joint cavity effusion culture were 76.9% (20/26), 100.0% (21/21), 100.0% (11/11), and 3/4, respectively. The combination therapy of doxycycline and rifampicin showed good therapeutic effects, and 22 patients were cured. Conclusions:Hainan is not an epidemic area of brucellosis, and the clinical manifestations of the disease are not specific. The epidemic and clinical characteristics of the disease should be paid attention to in order to improve the level of diagnosis and treatment and control effectiveness.

Objective:To study the epidemiological and clinical characteristics of patients infected with Streptococcus suis in Hainan Province. Methods:Clinical data of patients diagnosed with human infection with Streptococcus suis at the Second Affiliated Hospital of Hainan Medical University from January 2017 to December 2023 were collected, and their epidemiological characteristics, clinical manifestations, auxiliary examination results, treatment and prognosis were retrospectively analyzed. Results:A total of 29 patients infected with Streptococcus suis were included, with 26 males (89.66%) and 3 females (10.34%). The age of the patients was (54.52 ± 10.89) years old, with the highest number of patients aged 51 to 60 years, accounting for 41.38% (12 cases). Farmers were the main occupation (72.41%, 21/29). Forty-four point eighty-three percent (13/29) of the patients had a history of direct or indirect contact with dead or sick pigs. The number of cases from January to December was 0, 0, 2, 2, 4, 8, 2, 3, 2, 2, 3, and 1, respectively. June was the peak period of incidence (27.59%, 8/29). The main clinical manifestations were fever (100.00%, 29/29), altered state of consciousness (51.72%, 15/29), headache (48.28%, 14/29), and hearing impairment (44.83%, 13/29). In clinical classification, common type, meningitis type, and mixed type accounted for 20.69% (6/29), 75.86% (22/29), and 3.45% (1/29), respectively. In the pathogen examination, all 29 patients underwent blood culture, and 18 patients (62.07%, 18/29) had positive blood culture; 26 patients underwent cerebrospinal fluid culture, and 8 cases (30.77%) tested positive for cerebrospinal fluid culture. Eighty-nine point sixty-six percent (26/29) of the patients were treated with ceftriaxone sodium; 55.17% (16/29) of the patients were treated with combined treatment of ceftriaxone sodium and glucocorticoids, including 11 cases treated with dexamethasone and 5 cases treated with methylprednisolone. After treatment, 7 patients were cured and discharged, and 22 patients showed improvement in their condition and discharged from hospital. At discharge, both blood and/or cerebrospinal fluid cultures turned negative. Telephone follow-up was conducted 6 months after discharge, and 20 patients had no sequelae, 9 patients still had hearing impairment, including one who also developed walking difficulties. Conclusions:The patients infected with Streptococcus suis in Hainan Province are mainly middle-aged, elderly, male and farmers, with June being the peak period of the disease. The main clinical manifestations are fever, altered state of consciousness, headache, and hearing impairment. After follow-up, there are still patients with residual hearing impairment. In clinical practice, patients with fever and hearing impairment should be considered as this disease as early as possible.