This study aimed to investigate the effect and molecular mechanism of sinomenine on proliferation, apoptosis, metastasis, and combination with inhibitors in human hepatocellular carcinoma HepG2 cells and SK-HEP-1 cells. The effect of sinomenine on the growth ability of HepG2 and SK-HEP-1 cells were investigated by CCK-8 assay, colony formation assay, and BeyoClick~(TM) EdU-488 staining. The effect of sinomenine on DNA damage was detected by immunofluorescence assay, and the effect of sinomenine on apoptosis of human hepatocellular carcinoma cells was clarified by Hoechst 33258 staining and CellEvent~(TM) Cystein-3/7Green ReadyProbes~(TM) reagent assay. Cell invasion assay and 3D tumor cell spheroid invasion assay were performed to investigate the effect of sinomenine on the invasion ability of human hepatocellular carcinoma cells in vitro. The effect of sinomenine on the regulation of protein expression related to the protein kinase B(Akt)/mammalian target of rapamycin(mTOR)/signal transducer and activator of transcription 3(STAT3) signaling pathway in HepG2 and SK-HEP-1 cells was examined by Western blot. Molecular docking was used to evaluate the strength of affinity of sinomenine to the target cysteinyl aspartate specific proteinase-3(caspase-3) and STAT3, and combined with CCK-8 assay to detect the changes in cell viability after combination with STAT3 inhibitor JSI-124 in combination with CCK-8 assay. The results showed that sinomenine could significantly reduce the cell viability of human hepatocellular carcinoma cells in a concentration-and time-dependent manner, significantly inhibit the clonogenic ability of human hepatocellular carcinoma cells, and weaken the invasive ability of human hepatocellular carcinoma cells in vitro. In addition, sinomenine could up-regulate the cleaved level of poly ADP-ribose polymerase(PARP), a marker of apoptosis, and down-regulate the protein levels of p-Akt, p-mTOR, and p-STAT3 in human hepatocellular carcinoma cells. Molecular docking results showed that sinomenine had good affinity with the targets caspase-3 and STAT3, and the sensitivity of sinomenine to hepatocellular carcinoma cells was diminished after STAT3 was inhibited. Therefore, sinomenine can inhibit the proliferation and invasion of human hepatocellular carcinoma cells and induce apoptosis, and the mechanism may be attributed to the activation of caspase-3 signaling and inhibition of the Akt/mTOR/STAT3 pathway. This study can provide a new reference for the in-depth research and clinical application of sinomenine and is of great significance to further promote the scientific development and utilization of sinomenine.
Humans ; Carcinoma, Hepatocellular/genetics* ; Proto-Oncogene Proteins c-akt/metabolism* ; Caspase 3/metabolism* ; Liver Neoplasms/genetics* ; Molecular Docking Simulation ; Sincalide/pharmacology* ; Cell Line, Tumor ; Cell Proliferation ; Hep G2 Cells ; TOR Serine-Threonine Kinases/metabolism* ; Apoptosis

This study investigated the effect and mechanism of morin in inducing autophagy and apoptosis in hepatocellular carcinoma cells through the protein kinase B(Akt)/mammalian target of rapamycin(mTOR)/signal transducer and activator of transcription protein 3(STAT3) pathway. Human hepatocellular carcinoma SK-HEP-1 cells were stimulated with different concentrations of morin(0, 50, 100, 125, 200, and 250 μmol·L~(-1)). The effect of morin on the viability of SK-HEP-1 cells was detected by Cell Counting Kit-8(CCK-8). The effect of morin on the proliferation and apoptosis of SK-HEP-1 cells was investigated using colony formation assay, flow cytometry, and BeyoClick~(TM) EdU-488 with different concentrations of morin(0, 125, and 250 μmol·L~(-1)). The changes in the autophagy level of cells treated with morin were examined by transmission electron microscopy and autophagy inhibitors. The impact of morin on the expression levels of proteins related to the Akt/mTOR/STAT3 pathway was verified by Western blot. Compared with the control group, the morin groups showed decreased viability of SK-HEP-1 cells in a time-and concentration-dependent manner, increased number of apoptotic cells, up-regulated expression level of apoptosis marker PARP, up-regulated phosphorylation level of apoptosis-regulating protein H2AX, decreased number of positive cells and the colony formation rate, an upward trend of expression levels of autophagy-related proteins LC3-Ⅱ, Atg5, and Atg7, and decreased phosphorylation levels of Akt, mTOR, and STAT3. These results suggest that morin can promote apoptosis, inhibit proliferation, and induce autophagy in hepatocellular carcinoma cells, and its mechanism of action may be related to the Akt/mTOR/STAT3 pathway.
Humans ; Proto-Oncogene Proteins c-akt/metabolism* ; Carcinoma, Hepatocellular/pathology* ; Liver Neoplasms/pathology* ; TOR Serine-Threonine Kinases/metabolism* ; Apoptosis ; Autophagy ; Cell Proliferation ; Cell Line, Tumor ; STAT3 Transcription Factor/metabolism*

OBJECTIVE: To explore the amino acid metabolomics characteristics of myeloid-derived suppressor cells (MDSCs) in mice with sepsis induced by the cecal ligation and puncture (CLP). METHODS: The sepsis mouse model was prepared by CLP, and the mice were randomly divided into a sham operation group (sham group, n = 10) and a CLP model group (n = 10). On the 7th day after the operation, 5 mice were randomly selected from the surviving mice in each group, and the bone marrow MDSCs of the mice were isolated. Bone marrow MDSCs were separated to measure the oxygen consumption rate (OCR) by using Agilent Seahorse XF technology and to detect the contents of intracellular amino acids and oligopeptides through ultra-performance liquid chromatography/tandem mass spectrometry (UPLC-MS/MS) technology. Different metabolites and potential biomarkers were analyzed by univariate statistical analysis and multivariate statistical analysis. The major metabolic pathways were enriched using the small molecular pathway database (SMPDB). RESULTS: The proportion of MDSCs in the bone marrow of CLP group mice (75.53% ± 6.02%) was significantly greater than that of the sham group (43.15%± 7.42%, t = 7.582, P < 0.001), and the basal respiratory rate [(50.03±1.20) pmol/min], maximum respiration rate [(78.07±2.57) pmol/min] and adenosine triphosphate (ATP) production [(25.30±1.21) pmol/min] of MDSCs in the bone marrow of CLP group mice were significantly greater than the basal respiration rate [(34.53±0.96) pmol/min, (t = 17.41, P < 0.001)], maximum respiration rate [(42.57±1.87) pmol/min, (t = 19.33, P < 0.001)], and ATP production [(12.63±0.96) pmol/min, (t = 14.18, P < 0.001)] of sham group. Leucine, threonine, glycine, etc. were potential biomarkers of septic MDSCs (all P < 0.05). The increased amino acids were mainly enriched in metabolic pathways, such as malate-aspartate shuttle, ammonia recovery, alanine metabolism, glutathione metabolism, phenylalanine and tyrosine metabolism, urea cycle, glycine and serine metabolism, β-alanine metabolism, glutamate metabolism, arginine and proline metabolism. CONCLUSION The enhanced mitochondrial oxidative phosphorylation, malate-aspartate shuttle and alanine metabolism in MDSCs of CLP mice may provide raw materials for mitochondrial aerobic respiration, thereby promoting the immunosuppressive function of MDSCs. Blocking the above metabolic pathways may reduce the risk of secondary infection in sepsis and improve the prognosis.
Adenosine Triphosphate/metabolism* ; Alanine/metabolism* ; Animals ; Aspartic Acid/metabolism* ; Biomarkers/metabolism* ; Chromatography, Liquid ; Glycine/metabolism* ; Malates/metabolism* ; Mice ; Myeloid-Derived Suppressor Cells/metabolism* ; Sepsis/complications* ; Tandem Mass Spectrometry

OBJECTIVE: To observe and compare the clinical outcomes between arthroscopic modified Mason-Allen repair and suture-bridge repair for medium-size rotator cuff tears. METHODS: From January 2017 to January 2018, 22 patients with medium-size rotator cuff tears underwent arthroscopic modified Mason-Allen repair. There were 9 males and 13 females with an average age of (57.14±10.26) years. From February 2018 to January 2019, 20 patients with medium-size rotator cuff tears underwent arthroscopic suture-bridge repair. There were 6 males and 14 females with an average age of (57.75±7.57) years. The preoperative and postoperative clinical function was assessed by American Shoulder and Elbow Surgeons (ASES) and Constant score system. The healing status of repaired rotator cuff was assessed using MRI. RESULTS: All patients were followed up, and the duration ranged from 24 to 33 months, with a mean of (26.38±2.29) months. In modified Mason-Allen group, AS###ES score and Constant score increased from (45.22±7.58) and (58.72±9.26) preoperatively to (96.89±3.49) and (93.18± 3.20) postoperatively. In suture-bridge group, ASES score and Constant score increased from(47.33±7.50) and (60.05±11.76) scores to (97.58±3.43) and (93.85±3.15). There were no significant differences in ASES score and Constant score between the two groups before and after operation. There were no significant differences in rotator cuff healing between the two groups. CONCLUSION Both arthroscopic modified Mason-Allen and suture-bridge repair for treatment of medium-size rotator cuff tears could obtain good clinical outcomes, and there were no significant differences in clinical outcomes between the two techniques.
Aged ; Arthroscopy ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Rotator Cuff/surgery* ; Rotator Cuff Injuries/surgery* ; Suture Techniques ; Sutures ; Treatment Outcome

OBJECTIVES: To study the effect of glucose metabolism disorders on the short-term prognosis in neonates with asphyxia. METHODS: A retrospective analysis was performed on the medical data of the neonates with asphyxia who were admitted to 52 hospitals in Hubei Province of China from January to December, 2018 and had blood glucose data within 12 hours after birth. Their blood glucose data at 1, 2, 6, and 12 hours after birth (with an allowable time error of 0.5 hour) were recorded. According to the presence or absence of brain injury and/or death during hospitalization, the neonates were divided into a poor prognosis group with 693 neonates and a good prognosis group with 779 neonates. The two groups were compared in the incidence of glucose metabolism disorders within 12 hours after birth and short-term prognosis. RESULTS: Compared with the good prognosis group, the poor prognosis group had a significantly higher proportion of neonates from secondary hospitals (48.5% vs 42.6%, CONCLUSIONS Recurrent hyperglycemia in neonates with asphyxia may suggest poor short-term prognosis, and it is necessary to strengthen the early monitoring and management of the nervous system in such neonates.
Asphyxia ; Asphyxia Neonatorum/epidemiology* ; Humans ; Hyperglycemia ; Infant, Newborn ; Prognosis ; Retrospective Studies

Background: In China, secondary cytoreductive surgery (SCR) has been widely used in ovarian cancer (OC) over the past two decades. Although Gynecologic Oncology Group-0213 trial did not show its overall survival benefit in first relapsed patients, the questions on patient selection and effect of subsequent targeting therapy are still open. The preliminary data from our pre-SOC1 phase II study showed that selected patients with second relapse who never received SCR at recurrence may still benefit from surgery. Moreover, poly(ADP-ribose) polymerase inhibitors (PARPi) maintenance now has been a standard care for platinum sensitive relapsed OC. To our knowledge, no published or ongoing trial is trying to answer the question if patient can benefit from a potentially complete resection combined with PARPi maintenance in OC patients with secondary recurrence. Methods SOC-3 is a multi-center, open, randomized, controlled, phase II trial of SCR followed by chemotherapy and niraparib maintenance vs chemotherapy and niraparib maintenance in patients with platinum-sensitive second relapsed OC who never received SCR at recurrence. To guarantee surgical quality, if the sites had no experience of participating in any OC-related surgical trials, the number of recurrent lesions evaluated by central-reviewed positron emission tomography–computed tomography image shouldn't be more than 3. Eligible patients are randomly assigned in a 1:1 ratio to receive either SCR followed by 6 cyclesof platinum-based chemotherapy and niraparib maintenance or 6 cycles of platinum-based chemotherapy and niraparib maintenance alone. Patients who undergo at least 4 cycles of chemotherapy and must be, in the opinion of the investigator, without disease progression, will be assigned niraparib maintenance. Major inclusion criteria are secondary relapsed OC with a platinum-free interval of no less than 6 months and a possibly complete resection. Major exclusion criteria are borderline tumors and non-epithelial ovarian malignancies, received debulking surgery at recurrence and impossible to complete resection. The sample size is 96 patients. Primary endpoint is 12-month non-progression rate.

Objective To investigate the genetic variation and epidemiological characteristics of influenza B virus in Xinxiang to provide policy basis for local influenza vaccination. Methods The influenza surveillance data in Xinxiang from January 2012 to February 2019 was analyzed. 23 isolated influenza B virus were randomly selected for hemagglutinin (HA) and neuraminidase (NA) gene sequencing. Sequence alignment was conducted by using DNAman software and phylogenetic tree analysis was conducted using Neighbor-Joining method. Results Yamagata (BY) and Victoria (BV) strains of influenza B virus circulated alternately every other year in Xinxiang, mainly among people aged 0-15 years (91.4%). The dominant influenza B lineages from 2015 to 2016 and from 2017 to 2018 did not match the corresponding trivalent vaccine strain of the year. The HA phylogenetic tree revealed that 87.5% (7/8) of BV strains coexisted with the vaccine strain in one branch, while 88.98% (8/9) of BY strains from 2013 to 2015 were not in the same branch as the corresponding vaccine strain, with 5 epitope site mutions N116K, S150L, N165Y, D196N and N202S. No drug-resistant site mutation was identified in the NA gene. A total of 6 intra-lineage reassortants were identified. Conclusions The influenza B lineage in the trivalent vaccine recommended by WHO did not match the dominant circulating B lineage of Xinxiang in some epidemic year. Therefore, quadrivalent vaccines are recommended to use in susceptible population, especially under the age of 15. In addition, there are large variations in HA gene of the epidemic BY strain compared with the corresponding vaccine. Then, a vaccine more sutable for the epidemic strains in China is expected to be developed.

The disrupted caregiver infant relationships are not unpopular (infant abuse and non-traumatic caregiver-infant relationship).Animal and human studies have demonstrated this disruptive relationship exerts the fundamental and enduring impacts on stress system, limbic system and relevant cortex.Individual carrying such biological susceptibility might develop psychopathology under stresses at later life.The assessment needs to get involved the measures with more objectivity,real time and moment-to-moment components.The study on caregiverinfant relationship also should focus on the historical and cultural aspects of China.

BackgroundSeveral studies have investigated the association between smoking and anal abscess and anal fistula (AA/F) diseases. However, the relationship between cigarette smoking and AA/F remains unclear. This study sought to assess the role of smoking in anorectal male patients in a Chinese population.

MethodsIn this retrospective study, a questionnaire, including smoking history, was completed over a 3-month period by male inpatients in the Proctology Department of China-Japan Friendship Hospital. "Cases" were patients who had AA/F, and "controls" were patients with other anorectal complaints. Mann-Whitney U-test and Chi-square test were carried out to examine differences in baseline characteristics between groups. Subsequently, multivariate logistic regression was used to explore any related factors.

ResultsA total of 977 patients aged from 18 to 80 years were included, excluding those diagnosed with inflammatory bowel disease or diabetes mellitus. Out of this total, 805 patients (82.4%) completed the entire questionnaire. Among the 805 patients, 334 (41.5%) were cases and 471 (58.5%) were controls. Results showed significant differences between cases and controls (χ = 205.2, P < 0.001), with smoking found to be associated with the development of AA/F diseases (odds ratio: 12.331, 95% confidence interval: 8.364-18.179, P < 0.001).

ConclusionsThis study suggested smoking to be a potential risk factor for the development of AA/F diseases in a Chinese population. Consequently, current smoking patients should be informed of this relationship, and further research should be conducted to explore and investigate this further.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Anus Diseases ; epidemiology ; Female ; Humans ; Logistic Models ; Male ; Middle Aged ; Odds Ratio ; Rectal Fistula ; epidemiology ; Retrospective Studies ; Risk Factors ; Smoking ; adverse effects ; Surveys and Questionnaires ; Young Adult

Objective:The current study investigated the effects of nesfatin-1 in the hypothalamic paraventricular nucleus (PVN) on gastric motility and the regulation of the lateral hypothalamic area (LHA).Methods:The projection of nerve ?ber and expression of nesfatin-1 were observed by retrograde tracing and fluo-immunohistochemistry staining;The nuclei microinjection and nuclei electrical stimulation,extracellular discharges of single unit neuron were used to observe the effects of nesfatin-1 on the GD neurons;Gastric motility recording in vivo were used to monitor the effects of nesfatin-1 on the amplitude of constriction and frequency of gastric motility in conscious rats.Results:Nesfatin-1 inhibited the majority of the GD-E neurons(1.97± 0.12 Hz vs.1.15± 0.07 Hz) and excited GD-I neurons (1.74± 0.10 Hz vs.3.04± 0.18 Hz) in the PVN,which were weakened by oxytocin receptor antagonist H4928 (GD-E:1.38± 0.08 Hz,P＜0.05 vs.nesfatin-1;GD-I:2.49± 0.15 Hz,P＜0.05 vs.nesfatin-1).Gastric motility experiments showed that administration ofnesfatin-1 in the PVN decreased gastric motility.Retrograde tracing and immunofluorescent staining showed that nucleobindin-2/nesfatin-1 and fluorogold double-labeled neurons were observed in the LHA.Electrical LHA stimulation excited the firing rate of GD-responsive neurons (GD-E:2.06± 0.12 Hz vs.4.23± 0.21 Hz,GD-I:1.61± 0.09 Hz vs.4.83± 0.25 Hz) in the PVN.Pre-administration of an antinucleobindin-2/nesfatin-1 antibody in the PVN strengthened gastric motility,decreased GD-E neurons (1.74± 0.10 Hz vs.3.04± 0.18 Hz) and excited the discharging of the GD-I neurons(4.15± 0.18 Hz vs.4.83± 0.25) induced by electrical stimulation of the LHA.Conclusion:Nesfatin-1 in the PVN could serve as an inhibitory factor to inhibit gastric motility,which might be regulated by the LHA.