Changes in serum bone turnover markers and bone mineral density Z-score in children with osteogenesis imperfecta after zoledronic acid treatment.

The study aimed to investigate the changes in the levels of serum bone turnover markers (BTMs) and bone mineral density (BMD) Z-score in pediatric patients with osteogenesis imperfecta (OI) after intravenous bisphosphonate therapy and their association with age and estimated glomerular filtration rate (eGFR). This retrospective study analyzed data from 10 pediatric OI patients treated with intravenous zoledronic acid for over 1 year. Patients' clinical data were collected. The levels of BTMs and BMD Z-score before and after zoledronic acid treatment were analyzed. Significant improvement in BMD Z-score was observed after 6 and 12 months of treatment compared to baseline (all p < 0.05). The N-terminal propeptide of type I procollagen (PINP) levels decreased over time (all p < 0.05), indicating that zoledronic acid treatment decreased bone turnover. The levels of beta-C-terminal telopeptide of type I collagen remained stable after treatment. No correlation was found between PINP level and age, eGFR, or BMD (all p > 0.05). Bisphosphonate treatment can improve BMD and decrease bone turnover (indicated by decreased levels of PINP) in pediatric OI patients. PINP may serve as an independent indicator for monitoring the efficacy of bisphosphonate treatment in pediatric OI patients, particularly in those under the age of 6, where standardized BMD Z-score criteria are lacking.

Effect of Buzhong Yiqitang on Th17/Treg Immune Imbalance and Notch1 Signaling Pathway in AIT Mice / 中国实验方剂学杂志

ObjectiveTo explore the effect of Buzhong Yiqitang on the immune imbalance of helper T cell 17 （Th17）/regulatory T cell （Treg） and Notch1 signaling pathway in mice with autoimmune thyroiditis （AIT）. MethodA total of 60 8-week-old NOD.H-2h4 mice were randomly divided into the normal group， model group， western medicine group （selenium yeast tablet， 32.5 mg·kg-1）， and low-dose （4.78 g·kg-1·d-1）， middle-dose （9.56 g·kg-1·d-1）， and high-dose （19 g·kg-1·d-1） Buzhong Yiqitang groups， with 10 mice in each group. The normal group was fed with distilled water， and the other groups were fed with water containing 0.05% sodium iodide for eight weeks. After the animal model of AIT was formed spontaneously， the mice were killed under anesthesia after intragastric administration for eight weeks. Serum anti-thyroglobulin antibodies （TGAb）， thyroid-stimulating hormone （TSH）， free triiodothyronine （FT3）， and free thyroid hormone （FT4） were detected by enzyme-linked immunosorbent assay （ELISA）， and thyroid tissue changes were observed by hematoxylin-eosin （HE） staining. The mRNA and protein expressions of retinoid-related orphan receptor-γt （RORγt）， interleukin （IL）-17， forkhead box P3 （FoxP3）， IL-10， Notch1， and hair division-related enhancer 1 （Hes1） in thyroid tissue were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot， respectively. ResultCompared with the normal group， the thyroid structure of the model group was severely damaged， and lymphocytes were infiltrated obviously. The levels of serum TGAb， FT3， and FT4 contents were significantly increased， and TSH content was significantly decreased （P<0.01）. The mRNA and protein expression levels of RORγt， IL-17， Notch1， and Hes1 were significantly increased， while those of FoxP3 and IL10 were significantly decreased in the model group （P<0.01）. Compared with the model group， thyroid structural damage and lymphocyte infiltration were improved in the treatment groups， and serum TGAb， FT3， and FT4 contents were significantly decreased. TSH content was increased， and mRNA and protein expression levels of RORγt， IL-17， Notch1， and Hes1 were decreased. mRNA and protein expression levels of FoxP3 and IL-10 were increased to different degrees （P<0.05， P<0.01）， and the middle-dose Buzhong Yiqitang group had the most significant intervention effect. ConclusionBuzhong Yiqitang can alleviate the thyroid structural damage in AIT mice， and its mechanism may be related to improving the abnormal differentiation of Th17/Treg immune cells and inhibiting the activation of the Notch1 signaling pathway.

Clinical progress in the diagnosis and treatment of end-stage liver disease complicated with acute kidney injury / 中华全科医师杂志

Acute kidney injury (AKI) is a devastating complications of end-stage of liver disease (ESLD), seriously affecting the prognosis of patients. With the deepening understanding of the pathogenesis, the definition, staging, diagnosis and treatment of ESLD with AKI have been gradually optimized. This article reviews the evolution of definition, pathogenesis, diagnosis and treatment of ESLD with AKI, to provide reference for early recognition, precise diagnosis and standardized treatment of this condition.

Doping Copper Selenide for Tuning the Crystal Structure and Thermoelectric Performance of Germanium Telluride-Based Materials.

Germanium telluride (GeTe) compounds exhibit excellent thermoelectric performance. In this study, copper selenide (Cu2Se) was used to tune the crystal structure and carrier concentration (nH) of GeTe materials. The zT of the 1% Cu2Se-doped GeTe sample reaches 1.32, which is 52% higher than that of the pure phase. The results show that Cu2Se tunes the GeTe crystal structure and carrier concentration to achieve promising enhancements to the thermoelectric performance. Meanwhile, a herringbone-like crystal structure that reduces the lattice thermal conductivity was observed. However, because the directional movement of Cu ions at high temperatures leads to an increase in electrical conductivity, the electronic thermal conductivity also increased. This study focuses on crystal engineering strategies for the study of nontoxic thermoelectric materials.

Key Elements of Enhanced Recovery after Total Joint Arthroplasty: A Reanalysis of the Enhanced Recovery after Surgery Guidelines.

Recent guidelines have produced a consensus statement for perioperative care in hip and knee replacement. However, there is still a need for reanalysis of the evidence and recommendations. Therefore, we retrieved and reanalyzed the evidence of each recommended components of enhanced recovery after surgery (ERAS) based on the guidelines of total joint arthroplasty. For each one, we included for the highest levels of evidence and those systematic reviews and meta-analyses were preferred. The full texts were analyzed and the evidence of all components were summarized. We found that most of the recommended components of ERAS are supported by evidence, however, the implementation details of each recommended components need to be further optimized. Therefore, implementation of a full ERAS program may maximize the benefits of our clinical practice but this combined effect still needs to be further determined.

Advances in the protective mechanism and clinical implications of autophagy in liver failure / 临床肝胆病杂志

‍Liver failure is a serious clinical syndrome of liver disease with critical condition and high mortality， and besides liver transplantation， there is still a lack of satisfactory radical treatment methods. The pathogenesis of liver failure is complex and remains unclear， involving a variety of factors that affect the balance of hepatocyte necrosis and regeneration. This article summarizes autophagy as the key pathway for maintaining cell homeostasis and points out that autophagy plays an important protective role in the pathogenesis of liver failure by regulating NLRP3 inflammasome activation， reducing oxidative stress， and inhibiting cell apoptosis. Meanwhile， it is believed that the molecular signaling pathways targeting autophagy， such as exosomes and peroxisome proliferator－activated receptor α， participate in antagonizing the development and progression of liver failure and will become important ideas and directions for molecular targeted therapies for liver failure.

Research progress of lipoprotein a in cardiovascular diseases / 重庆医学

Lipoprotein a[Lp(a)]is a risk factor for cardiovascular disease.This paper summarizes the structure,anabolism and mechanism of action of Lp(a),and the relationship between Lp(a)and cardiovascular diseases,liver and kidney diseases,diabetes mellitus and other diseases.It focuses on the traditional lipid-low-ering regimen to reduce plasma Lp(a)level and the current popular novel therapies,such as mipomersen,pel-acarsen,olpasiran,and the interfering effect of related drugs on Lp(a)level and the degree of benefit on cardi-ovascular events.How to reduce plasma Lp(a)level and improve patient prognosis will be the key to future Lp(a)related research.

Type Ⅰ sialidosis:four case reports / 中国神经精神疾病杂志

Sialidosis is a rare autosomal recessive disorder which is classified into type Ⅰ and Ⅱ according to the age of onset and severity of the disease.Type I,also known as mucolipidosis,is often manifested as myoclonus,ataxia,general tonic clonic seizure,and bilateral cherry-red spots in eye examination.Four cases of type Ⅰ sialidosis were reported in order to improve the diagnosis and treatment of this disease.Patients were mainly manifested as myoclonus and ataxia.Whole exon gene sequencing of the four patients all showed the mutation c.544A＞G in the NEU1 gene.Meanwhile,the new mutation c.596G＞A carried was found in case 2 and its pathogenicity was also analyzed.Finally,we summarized the clinical characteristics of type Ⅰ patients in China,noting that most patients develop all the core symptoms as the disease progresses.

Enhancing bone regeneration: advances and innovations in exosome engineering / 中华骨科杂志

Effective bone repair and regeneration is crucial for treating skeletal tissue defects, including osteonecrosis, nonunion fractures, osteoporosis, and various other bone deficiencies. Exosomes, as cellular secretory vesicles, are pivotal in mediating intercellular communication through their cargo of proteins, lipids, and nucleic acids. Mesenchymal stem cell-derived exosomes, in particular, have emerged as promising agents in bone repair and regeneration, showing potential for practical application and clinical translation. Nonetheless, their functional capacity and therapeutic efficacy require enhancement. This review delineates exosome optimization strategies aimed at augmenting secretion and functionality, alongside the incorporation of exosome-functionalized biomaterials for bone healing. Evidence indicates that physical stimulation, molecular interventions, and small-molecule or biomaterial stimuli are effective in increasing exosome output. Moreover, engineering exosomes and their parental cells can further potentiate their therapeutic function. The amalgamation of exosomes with biomaterials represents a burgeoning approach in bone tissue engineering, offering novel therapeutic avenues. This comprehensive analysis aims to guide future applications and the clinical adoption of exosomes in bone tissue restoration.

The effects of Connexin-43 on osteoblasts cells proliferation and osteogenic differentiation / 中华骨科杂志

Objective:To investigate the effects of Connexin-43 (Cx43) on osteoblasts proliferation and osteogenic differentiation and its regulatory mechanism.Methods:Osteoblasts were isolated and cultured in vitro. The osteogenic activity of osteoblasts was detected by alizarin red staining and alkaline phosphatase (ALP) staining after dexamethasone treatment. The expression of Cx43, Runt-related transcription factor 2 (Runx2), ALP, collagen I type (COL-I) and proliferation-related proteins PCNA and CDK4 in osteoblasts were detected by Western-blot. The expressions of osteoblast proteins were detected by immunofluorescence staining. The proliferation of osteoblasts was detected by CCK8 assay. The lentivirus-mediated Cx43 gene overexpression plasmid (Lv-Cx43) was constructed and transfected into osteoblasts. The osteogenic activity and proliferation ability of osteoblasts were further detected by the above methods. Cx43 in osteoblasts was overexpressed by pretreating PD98059. The osteogenic activity and proliferation of Cx43 in overexpressed osteoblasts was detected by CCK8 and alizarin red staining.Results:The isolated osteoblasts have osteogenic differentiation ability. Compared with the control group, 1×10 -6 mol/L dexamethasone treatment could reduce the formation of calcium nodules in osteoblasts. With the increase of dexamethasone treatment duration, the protein expression of Cx43, Runx2, ALP and COL-I in osteoblasts decreased gradually, while the expression of PCNA, CDK4 and p-ERK1/2 decreased. The OD values of normal osteoblasts at 0, 1, 2, 3 and 4 d were 0.316±0.043, 0.891±0.623, 1.683±0.154, 2.315±0.721 and 2.891±0.323, respectively. However, The OD values of osteoblasts treated with dexamethasone were 0.376±0.021, 0.657±0.121, 1.124±0.285, 1.521±0.272, 1.987±0.584, respectively. OD values of dexamethasone treated osteoblasts were lower than those of normal group at 2, 3 and 4 days ( P<0.05). The relative expression levels of Cx43 mRNA in control group, Lv-NC group and Lv-Cx43 group were 0.541±0.086, 0.598±0.018 and 1.000±0.082, respectively. The mRNA expression level of Cx43 in Lv-Cx43 group was higher than that in control group and Lv-NC group ( P<0.05). The ratio of Cx43 protein band to the gray value of GAPDH band in control group, Lv-NC group and Lv-Cx43 group were 0.816±0.737, 0.738±0.643 and 1.145±1.101, respectively. The expression level of Lv-Cx43 was higher than that in control group and Lv-NC group ( P<0.05). The expressions of Runx2, ALP, COL-I mRNA and related marker proteins in Lv-Cx43 group were higher than those in control group and Lv-NC group ( P<0.05). The number of calcium nodules in the Lv-Cx43 group was significantly higher than that in the control group and Lv-NC group. The OD value of osteoblasts and the number of calcium nodules in Lv-Cx43+PD98059 group were significantly lower than those in Lv-Cx43 group ( P<0.05). Conclusion:The proliferation and differentiation ability of osteoblasts is significantly decreased after the treatment of dexamethasone with decreased expression of Cx43. Overexpression of Cx43 can promote the proliferation and osteogenic differentiation of osteoblasts, which may be regulated through the ERK1/2 pathway.

Research Progress on the Role of Mitochondrial DNA in the Pathogenesis of Chronic Liver Disease / 中华肝脏病杂志

Mitochondrial DNA is the mitochondria's own genetic material located within the mitochondrial matrix and is involved in cellular metabolism and energy supply. Mitochondrial DNA damage exacerbates oxidative stress by increasing the release of reactive oxygen species, while mitochondrial DNA release also triggers apoptosis and activates immune inflammatory responses through damage-related molecular patterns. Mitochondrial autophagy regulates mitochondrial DNA damage and release through a negative feedback mechanism to maintain intracellular homeostasis. Recent studies have shown that the occurrence and development of chronic liver disease are closely related to mitochondrial DNA-mediated immune inflammatory responses and oxidative stress.

Characteristics and trends of adolescents health risk behaviors in Shanghai from 2004 to 2019 / 中国学校卫生

Objective@#To analyze the trend of adolescent health risk behaviors in Shanghai, and to provide reference for the comprehensive intervention of middle school students health risk behaviors.@*Methods@#Based on the health risk behavior questionnaire of Chinese Center for Disease Control and Prevention, the questionnaire was adapted according to the actual monitoring needs. It was divided into junior high school version and senior high school version. From 2004 to 2019, using the method of multistage stratified cluster random sampling, 59 209 middle school students who completed the questionnaire in 6 surveys were selected for analysis. @*Results@#From 2004 to 2019, in the 7 days prior to the survey, 9.2%-50.6% of middle school students drank a glass of soda more than or equal to once a day and 54.2%-76.1% of middle school students reported eating dessert twice or more. Within 7 days, 48.3%-60.7% of middle school students had≥60 min of exercise per day for less than or equal to 3 days, and 16.1%- 35.2% of middle school students reported too much extracurricular activities, and the reporting rate increased year by year with the annual percent change ( APC ) of 5.15%( t =9.28, P <0.01). The reporting rate of long time online learning was 6.0%-13.6%, which showed an upward trend among high school students, with the APC of 5.35%( t =3.14, P <0.05). The reporting rate of middle school students pedestrian safety problems decreased from 69.1% in 2004 to 27.6% in 2019, with the APC of -6.28%( t =-8.18, P <0.01), but the reporting rate of cycling safety problems have increased in recent years. The reporting rate of intentional injury behaviors decreased by year such as fighting, bullying, etc. The reporting rate of initial smoking age ≤13 years old decreased, but attempted drinking behavior increased in junior middle school students, and the APC was 1.61%( t =3.48, P <0.05). A total of 1.6%-3.4% of middle school students had an Internet addiction behavior tendency. The detection rate of Internet addiction tendency was increasing in high school students and girls, and APC was 6.59% and 10.29% respectively( t =6.37, 8.62, P <0.01).@*Conclusion@#From 2004 to 2019, unintentional injury behavior, intentional injury behavior and substance addiction behavior of middle school students in Shanghai have improved. However, unhealthy diet and physical inactivity are still high. In the follow up. It need to focus on adverse diet and lack of physical activity behavior and take comprehensive intervention measures to control them.

Tranexamic acid attenuates early post-operative systemic inflammatory response and nutritional loss and avoids reduction of fibrinogen in total hip arthroplasty within an enhanced recovery after surgery pathway.

PURPOSE: Total hip arthroplasty (THA) is an intervention with significant inflammatory response. The impact of additional doses of tranexamic acid (TXA) on inflammatory response, trauma and nutrition parameters, and coagulation and fibrinolysis changes has rarely been reported. METHODS: A prospective double-blind randomized trial was performed on elective primary THA. Ninety-nine adult patients were recruited consecutively from 2019 to 2020. They were randomized to receive single-dose of TXA before incision, another dose of TXA at three hours post-operatively, or another two doses of TXA at three and six hours  post-operatively. The primary outcomes included changes in white blood cell (WBC) counts, creatine kinase (CK), haemoglobin(Hb), and albumin(Alb); the secondary outcomes included coagulation and fibrinolysis parameters. RESULTS: Compared with single-dose TXA, patients received three dose TXA had significantly reduced WBC counts and fibrinogen/fibrin degradation product (FDP) levels, increased albumin and fibrinogen levels, and prolonged PT on post-operative day (POD) three. Though patients received three dose TXA had a tendency that increased Hb, decreased CK, reduced D-D, and prolonged APTT on POD3, it is not statistically significant. And the other measured outcomes on POD1 and POD2W shared a similar statistical result, except PT. The PT is significantly prolonged on POD2W in three dose group compared with single dose. CONCLUSION: Three-dose TXA contribute to attenuate early post-operative systemic inflammatory response and nutritional loss, increase fibrinogen, reduce FDP levels, and prolong PT in THA patients within an ERAS pathway, which may associate with reduced early post-operative haemorrhagic tendency, thrombosis risks, and hypercoagulability.

Research Progress on the Relationship between Obesity-Inflammation-Aromatase Axis and Male Infertility.

Aromatase is a key enzyme in the transformation of androgen into estrogen. Its high expression will destroy the hormonal balance in the male body, and the excessive transformation of androgen into estrogen in the body will further damage the spermatogenic function of the testis, affect the normal development of the sperm, and cause spermatogenic disturbance. Adipose tissue has a high expression of aromatase and shows high enzymatic activity and ability to convert estrogen. Adipose tissue is the most estrogen-producing nongonadal tissue in the body because of its large size, accounting for about 20% of the body mass in healthy adults. PPARγ is recognized as the key adipose differentiation in the transcriptional regulation of the transcription factor. In the process of adipocyte differentiation, PPARγ regulate the expression of aromatase. The increase of aromatase is associated with the inflammatory response in adipose tissue caused by obesity. After obesity, the increase of proinflammatory factors in adipocytes will lead to enhanced transcription of the CYP19 gene encoding aromatase in adipocytes, which in turn will lead to increased expression of aromatase in adipocytes. This article reviews the regulation of male sterility from the angle of the "obesity-inflammation-aromatase" axis.

Survivorship and clinical outcomes of 'cup-cage' reconstruction in revision of hip arthroplasty for chronic pelvic discontinuity: A systematic review.

BACKGROUND: This study was conducted to assess the survivorship and clinical outcomes of cup-cage reconstruction technique in the revision of THA. METHODS: PubMed, OVID, EMBASE, and Cochrane Central Register of Controlled Trials (CENTRAL) up to February 2020 were searched. Studies that reported the clinical and radiological follow-up were identified. RESULTS: A total of 151 hips (145 patients) in six studies were included. The all-cause revision-free survivorship of cup-cage implant at the end of follow-up was 90.1% (136/151), with a mean follow-up of 64.4 months(range 12-135). The overall complication rate was 23.8% (36 of 151 hips), of which component problem, dislocation, infection and sciatic nerve palsy/injury were relatively common. All included studies reported improved clinical outcomes at the end of follow-up. CONCLUSION: Results suggested that revision of THA with a cup-cage has a favourable implant survivorship and clinical outcomes for the treatment of pelvic discontinuity, despite the high complications occurrence rates.

Study on the expression of connexin-43 in steroid-induced osteonecrosis of femoral head and osteoblasts in rats and its regulation mechanism / 中华骨科杂志

Objective:To investigate the expression of connexin-43 (Cx43) in steroid-induced osteonecrosis of femoral head and osteoblasts in rats and its regulation mechanism.Methods:The model of steroid-induced osteonecrosis of femoral head (SIONFH) of rat was established. Micro-CT and HE staining were used to observe the degree of bone trabecular destruction and the incidence of empty lacunae. The expression levels of Cx43 and PI3K/Akt signaling pathway related molecules and osteoblast-related proteins in model group and control group were detected by RT-PCT and Western blot. The osteoblast (OB) of rats was further isolated and cultured in vitro. Under treatment of dexamethasone (Dex), Cx43 expression in OB cells was detected by Western blot and immunofluorescence. Western blot was used to detect the effect of glucocorticoid (GC) on the expression of related molecules of PI3K/Akt/β-catenin signaling pathway. Akt activator (SC79) and PI3K inhibitor (LY294002) were used to study the molecular mechanism of Dex regulation on Cx43 expression in OB cells. The regulatory relationship between β-catenin and Cx43 was investigated by immunoprecipitation and small interfere RNA (siRNA) technology.Results:The model of SIONFH in rats was successfully established, which proved that Cx43 expression level in the SIONFH model group was significantly lower than that in the control group, and the expression level of Cx43 was positively correlated with the expression of PI3K/Akt signaling pathway related molecules and osteoblast-related proteins Runx2, ALP and Collagen I Type (COL). In addition, in vitro culture of isolated rat OB cells, the expression of Cx43, p-PI3K, P-Akt and β-catenin in OB cells decreased gradually as the Dex action time went on. Moreover, SC79 pretreatment could significantly reverse the inhibitory effect of GCs on Cx43 expression, while LY294002 could significantly enhance the inhibitory effect of GCs on Cx43. In addition, the immunoprecipitation results showed that β-catenin expression was closely related to Cx43 expression, and further studies showed that β-catenin-siRNA could significantly down-regulate the expression of Cx43.Conclusion:Under the action of GC, the expression level of Cx43 in bone tissue and OB cells decreased significantly, and the possible mechanism was that GCs inhibited the expression of Cx43 by inhibiting the PI3K/Akt/β-catenin signaling pathway, which laid a new theoretical foundation for the further study of the role of Cx43 in the pathogenesis of steroid-induced femoral head necrosis.

The patent situation analysis and countermeasure research of medical research institutions in Sichuan / 中华医学科研管理杂志

Objective:By analyzing the patent literature of medical research institutions in Sichuan Province, the current status of patent development in each institution were analyzed to identify existing problems, according to which countermeasures were put forward.Methods:Using IncoPat as the retrieval tool, the patent documents of Sichuan medical scientific research institutions were searched, data about the overall application trend, applicant, patent type, legal events were analyzed, and relevant countermeasures were proposed based on the current patent development situation.Results:At present, the overall development of patents in Sichuan medical scientific research institutions is good, however, there are still some problems and challenges, such as weak awareness of patent protection, low patent quality, insufficient transformation of patent achievements and so on.Conclusions:By analyzing the present situation of patent development of medical scientific research institutions in Sichuan Province, this study put forward some countermeasures, including strengthening publicity on intellectual property protection, improving the driving mechanism, reforming intellectual property management and achievement transformation mechanism, to provide reference for promoting the benign development of patents at medical scientific research institutions, as well as promote the development of medical science and technology innovation.

Genetic analysis of a juvenile with maturity-onset diabetes of the young type 12 / 中华医学遗传学杂志

OBJECTIVE@#To explore the genetic basis for a juvenile with maturity-onset diabetes of the young type 12(MODY12).@*METHODS@#High-throughput sequencing was carried out to screen for the variants. Candidate variant was verified by Sanger sequencing. Pathogenity of the variant was predicted by searching the genetic databases and analysis by using bioinformatic software.@*RESULTS@#Genetic testing indicated that the patient and his mother have both carried a heterozygous c.3976G>A variant (p.Glu1326Lys) in exon 32 of the ABCC8 gene. Prediction of the protein structure suggested the variant to be deleterious. Based on the guidelines of the American College of Medical Genetics and Genomics, the variant was predicted to be uncertain significance.@*CONCLUSION@#Whether the c.3976G>A variant of the ABCC8 gene is the cause of the disease in this patient or not depends on the functional studies and more case data. Above finding has enriched the spectrum of ABCC8 gene variants.

Effect of total saponins from Panax japonicus on non-alcoholic steatohepatitis by regulating autophagy / 中国中药杂志

Non-alcoholic steatohepatitis(NASH) was induced by high-sugar and high-fat diet in mice to investigate the intervention effect of total saponins from Panax japonicus(TSPJ) and explore its possible mechanism. Mice were fed with high-sugar and high-fat diet to establish NASH model, and intervened with different doses of TSPJ(15, 45 mg·kg~(-1)). The animals were fed for 26 weeks. The histomorphology and pathological changes of liver tissues were observed by HE staining. The transcriptional expression levels of miR-199 a-5 p, autophagy related gene 5(ATG5) and inflammatory cytokines interleukin-6(IL-6), interleukin-1β(IL-1β) and tumor necrosis factor α(TNF-α) in mouse liver were measured by quantitative Real-time polymerase chain reaction(qRT-PCR). Western blot was used to detect the expression of autophagy-related proteins ATG5, P62/SQSTM1(P62), and microtubule-associated protein light chain 3(LC3)-I/Ⅱ proteins in mouse liver. The expression of P62 protein was detected by immunofluorescence staining. In order to verify the targeting regulation relationship between miR-199 a-5 p and ATG5, miR mimic/inhibitor NC and miR-199 a-5 p mimic/inhibitor were transfected into Hepa 1-6 cells, and the expression of ATG5 mRNA and protein was detected. pMIR-reportor ATG5-3'UTR luciferase reporter gene plasmid was constructed and co-transfected with miR mimic/inhibitor NC and miR-199 a-5 p mimic/inhibitor into Hepa 1-6 cells to detect luciferase activity. In vivo, HE staining in the model group showed typical fatty degeneration and inflammatory infiltration, with increased expression of miR-199 a-5 p and decreased expression of ATG5 mRNA and protein. The expression of autophagy-associated protein P62 increased significantly, the ratio of LC3Ⅱ/Ⅰ decreased, and the transcriptional expression of inflammatory factors increased significantly. After the intervention by TSPJ, the pathological performance of liver tissue was significantly improved, the expression of miR-199 a-5 p decreased and the expression of ATG5 mRNA and protein increased, the expression of autophagy-associated protein P62 decreased significantly, the ratio of LC3Ⅱ/Ⅰ increased, and the transcriptional expression of inflammatory cytokines IL-6, IL-1β and TNF-α decreased significantly. In vitro, it was found that the expression of ATG5 mRNA and protein and luciferase activity decreased significantly in miR-199 a-5 p overexpression cells, while after inhibition of miR-199 a-5 p expression, the expression level of ATG5 mRNA and protein and luciferase activity increased. The results showed that TSPJ can improve NASH in mice fed with high-sugar and high-fat diet, and its mechanism may be related to the regulation of miR-199 a-5 p/ATG5 signal pathway, the regulation of autophagy activity and the improvement of inflammatory response of NASH.

Chikusetsu saponin Ⅳa ameliorates myocardial hypertrophy of rats through regulating expression of miR199a-5p/Atg5 / 中国中药杂志

The present study investigated the effects of chikusetsu saponin Ⅳa(CHS Ⅳa) on isoproterenol(ISO)-induced myocardial hypertrophy in rats and explored the underlying molecular mechanism. ISO was applied to establish a rat model of myocardial hypertrophy, and CHS Ⅳa(5 and 15 mg·kg~(-1)·d~(-1)) was used for intervention. The tail artery blood pressure was measured. Cardiac ultrasound examination was performed. The ratio of heart weight to body weight(HW/BW) was calculated. Morphological changes in the myocardial tissue were observed by HE staining. Collagen deposition in the myocardial tissue was observed by Masson staining. The mRNA expression of myocardial hypertrophy indicators(ANP and BNP), autophagy-related genes(Atg5, P62 and beclin1), and miR199 a-5 p was detected by qRT-PCR. Atg5 protein expression was detected by Western blot. The results showed that the model group exhibited increased tail artery blood pressure and HW/BW ratio, thickened left ventricular myocardium, enlarged myocardial cells, disordered myocardial fibers with widened interstitium, and a large amount of collagen aggregating around the extracellular matrix and blood vessels. ANP and BNP were largely expressed. Moreover, P62 expression was up-regulated, while beclin1 expression was down-regulated. After intervention by CHS Ⅳa at different doses, myocardial hypertrophy was ameliorated and autophagy activity in the myocardial tissue was enhanced. Meanwhile, miR199 a-5 p expression declined and Atg5 expression increased. As predicted by bioinformatics, Atg5 was a target gene of miR199 a-5 p. CHS Ⅳa was capable of preventing myocardial hypertrophy by regulating autophagy of myocardial cells through the miR-199 a-5 p/Atg5 signaling pathway.