Objective:To observe the effects of sacubitril/valsartan (LCZ696) on viral replication and cardiomyocyte apoptosis in mice with coxsackievirus B3 (CVB3)-induced viral myocarditis (VMC) and to analyze the underlying mechanisms.Methods:Forty BALB/c mice were randomly divided into four groups with 10 in each group: Sham, Sham+ LCZ696, VMC, and VMC+ LCZ696 groups. VMC model was established by intraperitoneal injection of 0.1 ml of CVB3 with a concentration of 10 6 TCID 50/ml into BALB/c mice, while the sham intervention was an equal volume of saline. The day of virus injection was defined as day 0. LCZ696 was administered by gavage at a dose of 60 mg/kg every day for seven consecutive days starting from day 1. Mouse survival rates were calculated. Echocardiography was used to evaluate the cardiac function of mice. The level of creatine kinase-MB (CK-MB) was detected by ELISA. Western blot was used to detect the levels of inflammatory cytokines (IL-6, TNF-α), apoptosis-related proteins (caspase-3, cleaved-caspase-3, Bax, Bcl-2), CVB3 surface protein (VP-1) and p-AKT/AKT in the hearts of mice. CVB3 mRNA in mouse hearts was measured by PCR. Inflammatory cell infiltration and cell apoptosis in mouse hearts were observed by HE staining and TUNEL staining, respectively. Results:Compared with the Sham group, the mice in the VMC group had a decreased survival rate and impaired cardiac function ( P<0.05). The levels of CK-MB, IL-6, TNF-α, cleaved-caspase-3/caspase-3, Bax/Bcl-2, VP-1, and CVB3 mRNA in the hearts of VMC mice increased significantly ( P<0.05), accompanied by increased expression of AKT, decreased phosphorylation of AKT ( P<0.05) and increased cell apoptosis. LCZ696 reversed the above changes. It could increase the survival rate, improve the cardiac function ( P<0.05), decrease cardiac inflammation, cell apoptosis and viral replication ( P<0.05), and increase the phosphorylation of AKT ( P<0.05). LCZ696 had no significant effects on the survival rate, cardiac function, myocardial injury, cardiac inflammation, cell apoptosis, viral replication or the expression of PI3K/AKT signaling pathway-related proteins in normal mice. Conclusions:LCZ696 could significantly inhibit cardiomyocyte apoptosis and reduce CVB3 replication in the hearts of VMC mice by regulating the PI3K/AKT pathway, thereby improving mouse cardiac function and survival rate.

BACKGROUND: Dysfunction of the gap junction channel protein connexin 43 (Cx43) contributes to myocardial ischemia/reperfusion (I/R)-induced ventricular arrhythmias. Cx43 can be regulated by small ubiquitin-like modifier (SUMO) modification. Protein inhibitor of activated STAT Y (PIASy) is an E3 SUMO ligase for its target proteins. However, whether Cx43 is a target protein of PIASy and whether Cx43 SUMOylation plays a role in I/R-induced arrhythmias are largely unknown. METHODS: Male Sprague-Dawley rats were infected with PIASy short hairpin ribonucleic acid (shRNA) using recombinant adeno-associated virus subtype 9 (rAAV9). Two weeks later, the rats were subjected to 45 min of left coronary artery occlusion followed by 2 h reperfusion. Electrocardiogram was recorded to assess arrhythmias. Rat ventricular tissues were collected for molecular biological measurements. RESULTS: Following 45 min of ischemia, QRS duration and QTc intervals statistically significantly increased, but these values decreased after transfecting PIASy shRNA. PIASy downregulation ameliorated ventricular arrhythmias induced by myocardial I/R, as evidenced by the decreased incidence of ventricular tachycardia and ventricular fibrillation, and reduced arrythmia score. In addition, myocardial I/R statistically significantly induced PIASy expression and Cx43 SUMOylation, accompanied by reduced Cx43 phosphorylation and plakophilin 2 (PKP2) expression. Moreover, PIASy downregulation remarkably reduced Cx43 SUMOylation, accompanied by increased Cx43 phosphorylation and PKP2 expression after I/R. CONCLUSION PIASy downregulation inhibited Cx43 SUMOylation and increased PKP2 expression, thereby improving ventricular arrhythmias in ischemic/reperfused rats heart.
Rats ; Male ; Animals ; Myocardial Reperfusion Injury/metabolism* ; Connexin 43/genetics* ; Sumoylation ; Down-Regulation ; Rats, Sprague-Dawley ; Arrhythmias, Cardiac/drug therapy* ; Myocardial Ischemia/metabolism* ; RNA, Small Interfering/metabolism*

BACKGROUND/OBJECTIVES: Sanghuangporus sanghuang (SS) has various medicinal effects, including anti-inflammation and anticancer activities. Despite the extensive research on SS, its molecular mechanisms of action on lung cancer are unclear. This study examined the impact of an SS alcohol extract (SAE) on lung cancer using in vitro and in vivo models.MATERIALS/METHODS: Different concentrations of SAE were used to culture lung cancer cells (A549 and H1650). A cell counting kit-8 assay was used to detect the survival ability of A549 and H1650 cells. A scratch assay and transwell cell invasion assay were used to detect the migration rate and invasive ability of SAE. Western blot analysis was used to detect the expression of B-cell lymphoma-2 (Bcl-2), Bcl2-associated X (Bax), cyclin D1, cyclin-dependent kinases 4 (CDK4), signal transducer and activator of transcription 3 (STAT3), and phosphorylated STAT3 (p-STAT3). Lung cancer xenograft mice were used to detect the inhibiting ability of SAE in vivo.Hematoxylin and eosin staining and immunohistochemistry were used to detect the effect of SAE on the structural changes to the tumor and the expression of Bcl-2, Bax, cyclin D1, CDK4, STAT3, and p-STAT3 in lung cancer xenograft mice. RESULTS: SAE could inhibit lung cancer proliferation significantly in vitro and in vivo without cytotoxicity. SAE suppressed the viability, migration, and invasion of lung cancer cells in a dose and time-dependent manner. The SAE treatment significantly decreased the proapoptotic Bcl-2/Bax ratio and the expression of pro-proliferative proteins Cyclin D1 and CDK4 in vitro and in vivo. Furthermore, SAE also inhibited STAT3 expression. CONCLUSIONS SAE reduced the cell viability and suppressed cell migration and invasion in human lung cancer cells. Moreover, SAE also exhibited anti-proliferation effects in vivo. Therefore, SAE may have benefits in cancer therapy.

Objective:To investigate the clinical, skeletal muscle pathological, and genetic characteristics of fatal infantile hypertonic myofibrillar myopathy (FIHMM).Methods:The clinical manifestations, laboratory assessments data and gene sequencing results of 10 patients diagnosed with FIHMM in Shenzhen Children′s Hospital from February 2017 to April 2021 were retrospectively analyzed.Magnetic resonance imaging (MRI) of both musculoskeletal system and the brain, and electromyogram (EMG) were performed in 3 cases, while muscle biopsy was performed in 2 cases.Results:Among these 10 cases, 1 case was from Northeast China and 1 case from East China, while the rest 8 cases were from South China.Eight of the 10 patients were male, and the other 2 cases were female.They were all born normal and not related to each other.The age of onset varied from 2 to 12 months.The main clinical manifestations for all the patients were progressive rigidity of the rectus abdominis (8 cases), neck muscles (7 cases), rectus abdominis (2 cases) and intercostal muscles (1 case), resulting in respiratory failure.Mildly to moderately elevated serum creatine kinase level was detected (436-5 804 IU/L) (reference range: 24-229 IU/L). Complex repetitive discharges can be seen in the EMG, without any myotonic potential.Muscle fiber degeneration, necrosis, and vacuolar degeneration were noted in the histopathological examination of the vastus lateralis and rectus abdominis.An abnormal red granular deposit was observed in a portion of the field of the modified Gomory Trichrome staining.Immunohistochemistry showed substantial deposition of desmin.Under the electron microscopy, the sarcomere structure of the muscle fibers was seriously disordered, with the destruction of Z-bands and the presence of granular deposits.The whole-exome sequencing identified the same homozygous variation c. 3G>A, p.Met1? of CRYAB gene in all the patients, but heterozygous variation in their parents. Conclusions:Axial muscles involvement, such as rectus abdominis rigidity, is the main clinical characteristic of FIHMM.c.3G>A, p.Met1? mutation in the CRYAB gene is a hotspot mutation in Chinese children.

Traumatic spinal cord injury (SCI) has devastating effects on patients′ physical and mental health. Autophagy is widely involved in various physiological and pathological processes of the body and plays a key role in spinal cord injury. The mammalian target of rapamycin (mTOR) is the main regulator of autophagy. mTOR regulation of autophagy is closely related to the pathological process of spinal cord injury, and can effectively promote the recovery of spinal cord injury. Therefore, mTOR is a promising target for treatment of spinal cord injury. This article reviews the role of mTOR signal transduction pathway in the regulation of autophagy in spinal cord injury, in order to provide reference for follow-up research.

Objective:To discuss the surgical efficacy of neuroendoscopic transsphenoidal approach for the removal of pituitary cystic lesion.Methods:Clinical data and efficacy of 32 patients with neuroendoscopic transsphenoidal surgery and pathological diagnosis of pituitary cystic lesion in the Affiliated Hospital of Jining Medical University from March 2013 to May 2019 were retrospectively analyzed.Results:Of the 32 patients, 29 patients were pathologically diagnosed with Rathke cysts and 3 patients with pituitary arachnoid cysts. The content of cyst could be completely removed and the relationship between cyst and sellarseptum and subarachnoid space could be clearly observed by using endoscopy. After followed-up for 0.5-1.0 year, headic, dizziness and visual impairment were improved. One patient relapsed, without serious complications or death.Conclusions:Transsphenoidal neuroendoscopic surgery is a safe and effective treatment for pituitary cystic lesion.

OBJECTIVE：To establish the method for determining protein binding rate of dipeptidyl peptidase- 4 inhibitor LGT- 6 in different species of plasma ，and to compare their difference. METHODS ：By equilibrium dialysis ，LGT-6（3，30，300，3 000 nmol/L）was equilibrated in rat ，monkey and human plasma （i. e. internal dialysis solution ）for 48 h，using phosphate buffer as the external dialysis solution. The concentration of LGT- 6 in internal and external dialysis solution was determined by UPLC-MS/MS using tolbutamide as internal standard ，and the plasma protein binding rate was calculated. The determination was performed on ACQUITY UPLC HSS T 3 column with water （containing 0.01％ formic acid ）-acetonitrile（containing 0.01％ formic acid ）as mobile phase at the flow rate of 0.6 mL/min. The column temperature was 40 ℃，and the sample size was 2 μL. The ion source was electrospray ion source ，and the multiple ion monitoring mode was used to carry out positive ionization scanning. The ion pairs for quantitative analysis were m/z 487.0→434.3（LGT-6），m/z 271.1→172.0（internal standard ），respectively. RESULTS ：At the concentrations of 3，30，300，and 3 000 nmol/L，the protein binding rates of LGT- 6 in rat plasma were （96.25±0.97）％，（84.16± 1.24）％，（78.25±0.61）％，（66.63±0.95）％；the protein protein binding rates in monkey plasma were （98.54±0.58）％，（87.27± 1.01）％，（79.35±0.86）％，（66.69±0.54）％；the protein binding rates in human plasma were （99.40±1.03）％，（84.48± 1.15）％，（77.62±0.77）％，（66.93±0.48）％. At the same concentration ，the protein binding rates of LGT- 6 in rat ，monkey and human plasma had no significant difference （P＞0.05）. In the same species of plasma ，there were significant differences in the plasma protein binding rates of different concentration of LGT-6 among those groups （P＜0.05），and it decreased with 才〔2016〕4015） the increase of drug concentration. CONCLUSIONS ： The method for the determination of plasma protein binding rate of LGT-6 is successfully established. The data revealed that the protein binding rate of LGT- 6 is concentration-dependent ， there was no obvious spec ies difference on protein binding rates of LGT- 6 in rat ，monkey and human plasma under the same concentration.

Objective To evaluate the imaging features of MRI in autoimmune pancreatitis (AIP) and analyze the diagnostic value before and after treatment.Methods MRI data of 20 AIP patients (14 males,6 females;average age:(54.0±10.3) years) from January 2013 to December 2015 were retrospectively analyzed.The sequences of MRI included T1 weighted imaging (WI),T2WI,MR cholangiopancreatography (MRCP),diffusion weighted imaging (DWI) and dynamic enhancement images.Fifteen of the patients received DWI again after hormone therapy.The location and extent of lesions,signal,patterns of dynamic enhancement,"pseudocapsule" sign and other accompanying signs,the apparent diffusion coefficient (ADC) of the lesion and the DWI manifestations before and after treatment were observed,calculated and compared.x2 test and paired t test were used to analyze the data.Results Five patients were confirmed by pathology and 15 by clinical follow-up.In MRI,16 patients appeared diffusive swollen pancreases,and 4 patients were with focal enlargement.Lesions of pancreas showed low signal on T1WI and high signal on T2WI.Twelve patients presented "pseudocapsule" around the lesions and progressive enhance was shown in the delayed phase on dynamic contrast enhanced MRI.Ten patients showed stenotic choledoch in the head of pancreas and segmented stenotic pancreatic duct in MRCP.All pancreatic lesions in the 10 patients presented high signals on DWI.The pancreatic morphology,the signal of the lesion,the "pseudocapsule" sign and the enhancement degree on MRI were significantly improved compared with those before treatment in 15 patients (x2 values:5.000-22.941,all P＜0.05).Mter hormone therapy,the signals on DWI were markedly weakened in pancreatic parenchyma,and ADC was significantly higher than that before treatment ((1.27± 0.14)×10-3 vs (1.05±0.16)×10-3 mm2/s;t=4.15,P＜0.01).Conclusion DWI could reflect the pathological and biological characteristics of AIP and be used to evaluate the therapeutic efficacy of hormone treatment in AIP.

Objective To establish an evaluation index system of electricity saving at general hospitals.Methods Based on civil building energy saving studies and in accordance with national regulations on hospital energy saving, the authors build an electricity saving index system for general hospitals.The indexes were reduced by the rough set theory, and their weight was determined by analytical hierarchy process and expert analysis.Results An electricity saving evaluation index system for general hospitals is so developed, consisting of six level-2 indexes and 27 level-3 indexes.Conclusions Such an evaluation index system can guide hospital electricity consumption and saving.

Objective:To observe the clinical effect of marzulene-S granules combined with phloroglucinol injections in the treat-ment of pregnant women with acute gastritis. Methods:Totally 102 cases of pregnant women with acute gastritis were randomly as-signed into two groups with 51 cases in each. The control group was treated with marzulene-S granules 30min before meals with warm water,0. 67g,tid,and the observation group was additionally given phloroglucinol injection 80 mg in 0. 9% sodium chloride injections 20ml,ivd,qd. The treatment course was 2 weeks. The clinical curative effect of the two groups was observed before and after the treat-ment,and the symptom score changes and symptom remission time were compared as well. Results:The effective rate of the observa-tion group was significantly higher than that of the control group(94. 12% vs 78. 43%,P<0. 05). After the treatment,the integral of epigastric pain,belching and dry mouth in the observation group was decreased significantly than that before the treatment(P<0. 05), and significantly lower than that in the control group(P<0. 05). The time of abdominal pain was(1. 8+0. 4)d and(3. 9+1. 1)d in the observation group and the control group,respectively,and the difference was statistically significant(P<0. 05). üo serious ad-verse reactions were found in the two groups. Conclusion:Marzulene-S granules combined with phloroglucinol injections in the treat-ment of pregnant women with acute gastritis is effective.