OBJECTIVES: To analyze the differentially expressed exosomal miRNAs in patients with chronic heart failure (CHF) complicated by hyperuricemia (HUA) and explore their potential as novel diagnostic molecular markers and their target genes. METHODS: This study was conducted among 30 CHF patients with HUA (observation group) and 30 healthy volunteers (control group) enrolled between September, 2020 and September, 2023. Peripheral blood samples were collected from 6 CHF patients with HUA for analyzing exosomal miRNAs by high-throughput sequencing, and the results were validated in the remaining 24 patients using qRT-PCR. GO and KEGG enrichment analyses were performed to predict the the target genes of the identified differential miRNAs. We also validated the differentially expressed miRNAs by animal experiment. RESULTS: A total of 42 differentially expressed exosomal miRNAs were detected in observation group by high-throughput sequencing; among them, miR-27a-5p was significantly upregulated (P=0.000179), and miR-139-3p was significantly downregulated (P=0.000058). In the 24 patients with both CHF and PUA, qRT-PCR validated significant upregulation of miR-27a-5p (P=0.004) and downregulation of miR-139-3p (P=0.005) in serum exosomes. When combined, miR-27a-5p and miR-139-3p had a maximum area under the curve (AUC) of 0.899 (95% CI: 0812-0.987) for predicting CHF complicated by HUA. GO and KEGG enrichment analyses suggested that the differential expressions of miR-27a-5p and miR-139-3p was associated with the activation of the AMPK-mTOR signaling pathway to activate the autophagic response. We obtained the same conclusion from animal experiment. CONCLUSIONS Upregulated exosomal miR-27a-5p combined with downregulated exosomal miR-139-3p expression can serve as a novel molecular marker for diagnosis of CHF complicated by HUA, and their differential expression may promote autophagy in cardiomyocytes by activating the AMPK-mTOR signaling pathway.
Humans ; Hyperuricemia/diagnosis* ; Heart Failure/genetics* ; MicroRNAs/metabolism* ; Exosomes/metabolism* ; Chronic Disease ; Male ; Female ; Middle Aged ; Animals

Objective To evaluate whether 6-gingerol(6G)can inhibit fibrosis and improve the cardiac and renal function and in rats with cardiorenal syndrome.Methods In the in vitro experiments of this study,the incorporation of isotope-labeled amino acids was used to detect the intervention of 6-gingerol on normal rat kidney-49F(NRK-49F)and normal rat kidney-52E(NRK-52E)cells.68 male SD rats weighing 200～250 g were used to establish a rat model of cardiorenal syndrome by ligating the left anterior descending coronary artery and performing 5/6 nephrectomy.The rats were randomly divided into a control group,a model group,a low dose 6-gingerol group(6 mg/kg),a high dose 6-gingerol group(30 mg/kg),and a losartan potassium group(20 mg/kg).The 6-gingerol group received intraperitoneal injection of 6-gingerol,while the control group and model group received intraperito-neal injection of an equal amount of physiological saline.The losartan group received oral administration of losartan potassium for a total of 6 weeks.After successful modeling,blood samples were taken for biochemical and cardiac ultrasound examinations.After the experiment,blood,heart,and kidney samples were taken for Masson,immuno-histochemistry,and Western blot.Results 6-gingerol 20 μmol/L can reduce NRK-49F collagen synthesis and inhibit NRK-52E protein synthesis.Biochemical results showed that the serum creatinine,urea nitrogen,and brain natriuretic peptide(BNP)levels of rats in the low and high dose 6-gingerol groups and the losartan group were all reduced,with high dose 6-gingerol groups and losartan group showing the most significant decrease(P<0.05).Echocardiographic parameters showed that the 6-gingerol group and losartan potassium group improved cardiac contractile function and ventricular remodeling in rats(P<0.05).Masson staining and Western Blot showed renal collagen deposition,with reduced expression of collagen I and α-SMA(P<0.05).Immunofluorescence showed a decrease in the expression of renal collagen deposition I,α-SMA,and TGF-β1(P<0.05).Conclusion 6-Gin-gerol may improve the cardiac and renal function and renal fibrosis in rats with cardiorenal syndrome.

Objective:To investigate the effects of chronic persistent hypoxia on hepatic function, histological morphology, and ultrastructure in aged mice, and to evaluate the protective role of pyrroloquinoline quinone(PQQ).Methods:Thirty-two 2-month-old (young group)and thirty-two 18-month-old(aged group)male C57BL6/J mice were each randomly divided into four groups (n=8 per group): normoxia+ normal saline (NS)group, normoxia+ PQQ group, hypoxia+ NS group, and hypoxia+ PQQ group.The normoxia+ NS and normoxia+ PQQ groups were housed under normoxic conditions[fraction of inspired oxygen(FiO 2)=21%], while the hypoxia+ NS and hypoxia+ PQQ groups were continuously exposed to a hypoxic environment[FiO 2=(10±0.5)%]simulated by a custom-made hypoxic chamber, maintaining a constant oxygen concentration for 24 hours per day.The normoxia+ NS and hypoxia+ NS groups received daily intragastric administration of NS, whereas the normoxia+ PQQ and hypoxia+ PQQ groups received daily intragastric administration of PQQ disodium salt(8 mg·kg -1·d -1).After 8 weeks of continuous intervention, blood samples were collected to measure red blood cell count, hemoglobin levels, and liver function-related biochemical indicators.Lung tissues were processed for HE staining, and liver tissues were processed for both HE staining and electron microscopy.The histological and ultrastructural features of each group were observed under light and electron microscopy, respectively, and the differences between the groups were compared and analyzed. Results:Compared with the normoxia+ NS groups, both young and aged hypoxia+ NS groups exhibited significant pulmonary arteriole narrowing( P<0.001), with markedly elevated red blood cell count and hemoglobin levels (all P<0.001), which were not alleviated by PQQ.Compared with the young normoxia+ NS group, the young hypoxia+ NS group showed significantly higher alanine aminotransferase (ALT)and lactate dehydrogenase (LDH)levels( Z=2.72, 2.53, P=0.007, 0.011), whereas the young hypoxia+ PQQ group exhibited LDH levels similar to those of the young normoxia+ NS group.The aged hypoxia+ NS group exhibited significant ALT elevation( t=2.66, P=0.013)compared with the aged normoxia+ NS group.Light microscopy revealed hepatocyte ballooning degeneration, mild fatty accumulation, and focal necrosis around central veins in the young hypoxia+ NS group, while the young hypoxia+ PQQ group exhibited no significant pathological damage but displayed numerous deeply stained binucleated hepatocytes.The aged normoxia+ NS group demonstrated hepatocyte ballooning degeneration and inflammatory cell infiltration around central veins, whereas the aged normoxia+ PQQ group exhibited no obvious pathological damage with scattered deeply stained binucleated hepatocytes.The aged hypoxia+ NS group exhibited significant necrosis following physiological oxygen concentration gradient distribution, while the aged hypoxia+ PQQ group displayed no obvious pathological damage with scattered deeply stained binucleated hepatocytes.Electron microscopy revealed that the aged normoxia+ NS group had reduced mitochondrial electron density ( P<0.001)and less developed rough endoplasmic reticulum compared with the young normoxia+ NS group.The young hypoxia+ NS group exhibited a smaller mitochondrial area( P<0.001), decreased mitochondrial matrix electron density( P<0.001), blurred or absent mitochondrial cristae, inactive rough endoplasmic reticulum, and increased accumulation of glycogen and lipid droplets compared with the young normoxia+ NS group, while the young hypoxia+ PQQ group maintained mitochondrial matrix electron density comparable to the young normoxia+ NS group.The aged hypoxia+ NS group exhibited larger mitochondrial area( P=0.001), decreased mitochondrial matrix electron density( P<0.001), blurred or absent mitochondrial cristae, mitochondrial edema, increased lysosomes, and elevated cytoplasmic electron density compared with the aged normoxia+ NS group.The aged hypoxia+ PQQ group exhibited reduced mitochondrial area( P<0.001)and restored mitochondrial matrix electron density to levels comparable with the aged normoxia+ NS group.The aged normoxia+ PQQ group demonstrated increased mitochondrial matrix electron density compared with the aged normoxia+ NS group( P<0.001). Conclusions:Chronic persistent hypoxia induces hepatic functional, histological and ultrastructural damage in mice, with more pronounced effects in aged animals.PQQ provides a certain degree of protection against these injuries.

BACKGROUND:In recent years,the application of exosomes of periodontal ligament stem cells in periodontal tissue regeneration engineering has been widely studied,but the effect of exosomes on periodontal ligament stem cells derived from inflammatory environment is still unclear.OBJECTIVE:To investigate the effects of exosomes secreted by periodontal ligament stem cells from healthy and inflammatory environments on the proliferation and differentiation of periodontal ligament stem cells from inflammatory environments.METHODS:Human periodontal ligament stem cells from healthy and inflammatory tissues were isolated and cultured by enzyme digestion method.Exosomes were extracted from two kinds of periodontal ligament stem cells using ultracentrifugation.Passage 3 periodontal ligament stem cells derived from inflammatory tissue were selected and cultured in three groups.Cells in the blank group were cultured routinely.The healthy exosome group was added with exosomes secreted by peripheral ligament stem cells derived from healthy tissue.The inflammatory exosome group was added with exosomes secreted by human periodontal ligament stem cells derived from inflammatory tissue.Cell proliferation and cloning were detected.The expression of alkaline phosphatase,the formation of mineralized nodules,and the expression of mRNA and protein of genes related to osteogenesis were detected under osteogenic differentiation.RESULTS AND CONCLUSION:(1) CCK-8 assay and clonal formation test showed that compared with the blank group,two kinds of exosomes could promote the proliferation and colony formation of periodontal ligament stem cells from inflammatory tissue (P<0.05),and the effect of the healthy exosome group was stronger than that of the inflammatory exosome group (P<0.05).(2) Alkaline phosphatase and alizarin red staining showed that compared with the blank group,the two kinds of exosomes could promote the expression of alkaline phosphatase and the formation of mineralized nodules in periodontal ligament stem cells from inflammatory tissue,and the promoting effect of the healthy exosome group was stronger than that of the inflammatory exosome group.RT-PCR and western blot assay showed that compared with the blank group,the two kinds of exosomes could promote the expression of alkaline phosphatase,RUNX2,and type Ⅰ collagen mRNA and protein in periodontal ligament stem cells from inflammatory tissue (P<0.05).The promoting effect of the healthy exosome group was stronger than that of the inflammatory exosome group (P<0.05).(3) The results showed that exosomes secreted by human periodontal ligament stem cells could promote the proliferation and osteogenic differentiation of periodontal ligament stem cells derived from inflammatory environments,and the promoting effect of exosomes secreted by human periodontal ligament stem cells derived from healthy tissues was better than that from human periodontal ligament stem cells derived from inflammatory tissues.

BACKGROUND:Deep muscle stimulation has the effects of releasing muscle adhesion,relieving muscle spasm,improving and restoring muscle compliance and elasticity.Electromyographic biofeedback therapy can promote nerve recovery and improve lower limb motor function and gait. OBJECTIVE:To observe the effect of the effect of deep muscle stimulation combined with electromyographic biofeedback therapy on the spasm of the triceps surae and gait changes after stroke by using a digital muscle detector and three-dimensional gait analysis system. METHODS:A total of 72 patients who met the inclusion criteria were selected from the Rehabilitation Department of the First Affiliated Hospital of Guangdong Pharmaceutical University from October 2020 to October 2023.And they were enrolled and randomly divided into two groups(n=36 per group):a control group and a combined group.The control group received routine rehabilitation therapies,electromyographic biofeedback and pseudo deep muscle stimulation,while the combined group received true deep muscle stimulation treatment on the basis of the control group,five times per week,for 4 consecutive weeks.The oscillation frequency and dynamic stiffness of the affected gastrocnemius muscle,active range of motion of the ankle dorsiflexion muscle,electromyographic signal of the tibialis anterior muscle,Fugl-Meyer assessment of the lower limbs,and three-dimensional gait analysis parameters were statistically analyzed before and after treatment in two groups. RESULTS AND CONCLUSION:After treatment,oscillation frequency and dynamic stiffness values of the inner and outer sides of the affected gastrocnemius muscle in both groups of patients were significantly reduced compared with before treatment(P<0.05),and the combined group showed a more significant decrease compared with the control group(P<0.05).The active range of motion of the ankle dorsiflexion muscle,electromyographic signal of the tibialis anterior muscle,and Fugl-Meyer scores after treatment were significantly increased or improved compared with before treatment(P<0.05),while the combined group showed a more significant increase or improvement compared with the control group(P<0.05).In terms of gait parameters,the walking speed,frequency,and stride in both groups of patients were significantly increased compared with before treatment(P<0.05),while the combined group showed a more significant increase compared with the control group(P<0.05).The percentage time of support phase on the healthy side was shortened compared with before treatment(P<0.05),while the combined group showed a more significant decrease compared with the control group(P<0.05).In addition,there was no significant difference between the two groups except for the percentage of healthy side support(P>0.05).To conclude,the combination of deep muscle stimulation and electromyographic biofeedback can effectively alleviate triceps spasm in the short term after stroke,improve ankle dorsiflexion function,enhance lower limb motor function,and improve gait.The treatment effect is significant and worthy of clinical promotion and application.

As new evidence emerges, treatment strategies toward the functional cure of chronic hepatitis B are evolving. In 2019, a panel of national hepatologists published a Consensus Statement on the functional cure of chronic hepatitis B. Currently, an international group of hepatologists has been assembled to evaluate research since the publication of the original consensus, and to collaboratively develop the updated statements. The 2.0 Consensus was aimed to update the original consensus with the latest available studies, and provide a comprehensive overview of the current relevant scientific literatures regarding functional cure of hepatitis B, with a particular focus on issues that are not yet fully clarified. These cover the definition of functional cure of hepatitis B, its mechanisms and barriers, the effective strategies and treatment roadmap to achieve this endpoint, in particular new surrogate biomarkers used to measure efficacy or to predict response, and the appropriate approach to pursuing a functional cure in special populations, the development of emerging antivirals and immunomodulators with potential for curing hepatitis B. The statements are primarily intended to offer international guidance for clinicians in their practice to enhance the functional cure rate of chronic hepatitis B.

ObjectiveTo analyze the incidence and trend of stroke in Beilun District, so as to provide evidence for identifying influencing factors and reducing stroke incidence. MethodsStroke cases from 2012 to 2023 were extracted from the Ningbo Chronic Disease Collaborative Management System. Population information of Beilun District during the same period was also collected. The annual incidence and trends of stroke were analyzed. ResultsFrom 2012 to 2023, the age-standardized incidence rate of stroke in Beilun District, Ningbo City was 317.68/100 000, showing an increasing trend with an average annual percentage change (AAPC) of 2.267% (P=0.034). Among all subdistricts in Beilun District, two showed a downward trend in incidence, while the rest showed an upward trend. The crude incidence rate of stroke was significantly higher in males than that in females (P<0.001). The age-standardized incidence rate in males was 406.08/100 000, showing an increasing trend (AAPC=3.956%, P<0.001). The incidence of stroke also showed an increasing trend in the following age groups: 30‒<45 years (AAPC=6.340%, P=0.004), 45‒<60 years (AAPC=4.997%, P<0.001), and 60‒<75 years (AAPC=3.282%, P=0.042). Across all years, males had higher crude incidence rates in both ischemic and hemorrhagic stroke than females (P<0.05). The age-standardized incidence rate of ischemic stroke showed a rising trend in both males and the general population (male AAPC=4.905%, P<0.001; overall population AAPC=3.065%, P=0.001). ConclusionThe age-standardized incidence of stroke in Beilun District is on the rise, with higher crude incidence rate in males than that in females. The onset age of stroke is gradually declining. The age-standardized incidence rate of male ischemic stroke shows a clear upward trend.

OBJECTIVE: To investigate the effect of relaxing needling at the contracted sites of meridian-muscle regions in the patients with post-stroke shoulder-hand syndrome (SHS) at acute stage. METHODS: Eighty patients with post-stroke SHS at acute stage were randomized into an observation group (40 cases, 1 case dropped out) and a control group (40 cases, 1 case was eliminated). In the control group, the routine medication, basic rehabilitation training, and hyperbaric oxygen therapy were administered. In the observation group, besides the treatment as the control group, relaxing needling was delivered at the contracted sites of meridian-muscle regions. These contracted sites were distributed along three yin meridians of hand and three yang meridians of hand on the affected upper limbs. The intervention was given once daily, 5 times a week and for 4 weeks. Before and after treatment, the scores of visual analogue scale (VAS) for pain, edema degree, modified Barthel index (MBI), and Fugl-Meyer assessment (FMA) for motor function, and the integrated electromyography (iEMG) of surface electromyogram (sEMG) were observed in the two groups. The curative effect was evaluated after treatment and in follow-up of 2 months after treatment in the two groups. RESULTS: After treatment, VAS scores and the scores of edema degree were reduced when compared with those before treatment in the two groups (P<0.05), and the scores in the observation group were lower than those in the control group (P<0.05). MBI and FMA scores increased after treatment compared with those before treatment in the two groups (P<0.05), and the scores in the observation group were higher than those in the control group (P<0.05) after treatment. The iEMG values of the biceps brachii, triceps brachii, and wrist extensors were elevated after treatment in comparison with those before treatment (P<0.05) in the two groups, and the values in the observation group were larger than those in the control group after treatment (P<0.05). The total clinical effective rate in the observation group was 92.3% (36/39), which was better than that of the control group (74.4%, 29/39, P<0.05) after treatment; and that of the observation group was 97.4% (38/39), which was better than 82.1% (32/39) in the control group (P<0.05) in follow-up. CONCLUSION Relaxing needling at the contracted sites of meridian-muscle regions in treatment of post-stroke SHS at acute stage can attenuate the symptoms such as upper limb pain, swelling and spasm, improve motor function and the activity of daily living of patients.
Humans ; Male ; Female ; Middle Aged ; Acupuncture Therapy ; Aged ; Meridians ; Stroke/complications* ; Reflex Sympathetic Dystrophy/etiology* ; Adult ; Acupuncture Points

Objective To investigate the clinical efficacy of Tongfu Xiefei Formula in treating patients with acute exacerbation of chronic obstructive pulmonary disease(AECOPD)based on the theory of simultaneous treatment of lung and intestine,and to observe its effects on serum inflammatory cytokines.Methods A retrospective study was conducted on 134 AECOPD patients treated at the Department of Pulmonary Diseases of Traditional Chinese Medicine,Tangshan Hospital of Traditional Chinese Medicine from December 2020 to December 2022.The patients were divided into an observation group and a western medicine group based on the treatment plans,with 67 cases in each group.The western medicine group received conventional western medical treatment,while the observation group was given Tongfu Xiefei Formula orally in addition to the western medical treatment.The course of treatment covered 2 weeks.Before and after treatment,the two groups were observed in the changes of the modified Medical Research Council(mMRC)dyspnea scale scores,COPD Assessment Test(CAT)scores,lung function indicators,arterial blood gas analysis indicators,and serum inflammatory cytokine levels.The clinical efficacy,total incidence of adverse reactions,and hospitalization time were compared between the two groups.Results(1)After 2 weeks of treatment,the total effective rate in the observation group was 95.52%(64/67),compared to 79.10%(53/67)in the western medicine group.The intergroup comparison(tested by the chi-square test)showed that the efficacy of the observation group was significantly superior to that of the western medicine group(P＜0.01).(2)After treatment,the mMRC scores and CAT scores in both groups were significantly decreased(P＜0.05),and the decrease in the observation group was significantly superior to that in the western medicine group(P＜0.01).(3)After treatment,lung function indicators of the forced expiratory volume in one second(FEV1),forced vital capacity(FVC),and their ratio(FEV1/FVC)in both groups were significantly increased(P＜0.05),and the increase in the observation group was significantly superior to that in the western medicine group(P＜0.01).(4)After treatment,the oxygen saturation(SaO2)and arterial oxygen partial pressure(PaO2)levels in both groups significantly increased(P＜0.05),while the arterial carbon dioxide partial pressure(PaCO2)level was significantly decreased(P＜0.05).The increase in SaO2 and PaO2 levels and the decrease in PaCO2 level in the observation group were significantly superior to those in the western medicine group(P＜0.01).(5)After treatment,the levels of serum inflammatory cytokines of tumor necrosis factor α(TNF-α),C-reactive protein(CRP),and interleukin-6(IL-6)in both groups were significantly decreased(P＜0.05),and the decrease in the observation group was significantly superior to that in the western medicine group(P＜0.01).(6)The total incidence of adverse reactions in the observation group was 2.99%(2/67),compared to 5.97%(4/67)in the western medicine group,with no statistically significant difference between the two groups(P＞0.05).(7)The observation group had a significantly shorter hospitalization time than the western medicine group,and the difference was statistically significant(P＜0.05).Conclusion Tongfu Xiefei Formula,formulated based on theory of simultaneous treatment of lung and intestine,is effective and safe on relieving symptoms such as dyspnea in AECOPD patients,improving lung function,correcting arterial blood gas disorders,inhibiting the release of inflammatory factors,shortening treatment time,while causing no serious adverse reactions.

Abstract The incidence of fragility fractures and related mortality caused by male osteoporosis (OP) are both higher than in females. Male bone health was regulated by a sophisticated network involving both androgens and estrogens. Androgens can directly promote bone formation and inhibit bone resorption through androgen receptors on osteoblasts and osteocytes. They can also be converted into estrogens via the action of aromatase and subsequently regulate bone metabolism through estrogen receptors. Declining sex hormone levels, an imbalance in the estrogen-to-androgen ratio, and the resulting disruption in bone metabolic pathways collectively contribute to the development and progression of male OP. Moreover, androgens cannot fully compensate for the bone metabolic imbalance induced by estrogen deficiency. This article reviewed research on the role and mechanism of sex hormones in male OP, as well as studies related to the risks of sex hormone therapy for OP, so as to provide the references for improving control and treatment strategies of male OP.