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 analyze the reported characteristics of pulmonary tuberculosis (TB) in children aged 0-14 years in four provinces (municipalities), Beijing, Hubei, Chongqing and Sichuan, in China, and provide evidence for the prevention and control of pulmonary TB in children. Methods: The incidence data of childhood pulmonary TB were collected from notifiable disease and tuberculosis management information system of Chinese information system for disease control and prevention,and descriptive epidemiological methods were used to analyze the medical care seeking flow, characteristics and management inclusion of pulmonary TB cases in children. Statistical analysis and data visualization were conducted with softwares Excel 2015, R 4.1.2 and Echart 4.7.0. Results: A total of 6 811 pulmonary TB cases in children were reported in the four provinces during 2019-2021, in which 4 741 (69.6%) were clinically diagnosed and 2 070 (30.4%) were laboratory confirmed. A total of 526 medical institutions reported TB cases in children, including 356 general hospitals (67.7%, 356/526) reporting 4 706 cases, 11 infectious disease hospitals (2.1%, 11/526) reporting 836 cases and 5 children's hospitals (1.0%, 5/526) reporting 542 cases. A total of 6 249 (91.7%) local cases and 562 (8.3%) non-local cases were reported. The reported local incidence rates of TB from 2019-2021 were 6.20/100 000, 7.10/100 000 and 7.20/100 000, respectively, showing an increase trend year by year. The sex ratio of the cases were 0.98∶1(3 373∶3 438). The cases were mainly distributed in age group 10-14 years (4 887 cases, 71.8%). The cases were mainly students (5 167 cases, 75.9%). The management inclusion rates of the local cases and non-local cases were 20.60% and 2.67%, respectively. Conclusions: The main medical institutions reporting pulmonary TB cases in children were children's hospitals, infectious disease hospitals and TB special hospitals, the incidence of pulmonary TB in children in Sichuan was higher. In 2020, the inter-provincial medical seeking behavior of the pulmonary TB cases decreased significantly. The incidence rate in boys was lower than that in girls, and children aged 10-14 years were the population with high incidence of pulmonary TB. The management inclusion rate in non-local cases was lower than that in local cases.
Male ; Child ; Female ; Humans ; Tuberculosis, Pulmonary/epidemiology* ; Hospitals, Chronic Disease ; China/epidemiology* ; Asian People ; Hospitals, General

OBJECTIVES: To explore the development process of the postmortem interval (PMI) research in China from January 1990 to August 2020, research hotspots in different periods, authors and cooperation between institutions, and to provide a reference for the better development of PMI inference research through the visualization of the literature information of the PMI estimation research indexed in China National Knowledge Infrastructure (CNKI). METHODS: The information visualization analysis software CiteSpace 5.7.R1 was used to carry out big data analysis on hotspots, high-frequency keywords, authors, institutions and other information in the research literature on PMI inference from January 1990 to August 2020 indexed in CNKI. RESULTS: The peak time of publication of PMI was from 2006 to 2010 with 114 articles. In keyword co-occurrence network, the effective hot words were forensic entomology, DNA content analysis and some emerging words such as artificial intelligence and big data. In the cooperation network of institutions, the high-frequency institutions were mainly the scientific research institutions. The author cooperation network showed a trend of co-aggregation and multi-cooperation. CONCLUSIONS With the development of science and technology, the research on PMI estimation based on traditional methods is mature and novel strategies are emerging. Big data and artificial intelligence combined with forensic science provide new research directions on PMI estimation.
Artificial Intelligence ; Autopsy ; China ; Forensic Sciences ; Software

AIM:To analyze the imageological changes of acute and chronic central serous chorioretinopathy (CSC) by 2 types of optical coherence tomography (OCT). METHODS:A retrospective analysis was performed, inclu-ding data of 60 eyes from 56 patients with CSC diagnosed by conventional eye examination, fundus fluorescein angiography (FFA) and indocyanine green angiography (ICGA), which were divided into acute group (28 eyes of 28 patients) and chronic group (32 eyes of 28 patients) according to imageological examinations and duration (6 months). Optical coher-ence tomography angiography (OCTA) and spectral domain optical coherence tomography ( SD-OCT) were performed to study the vessel density of the chorioretinal leyers and the integrity of the outer retinal structure. RESULTS:In the pa-tients with chronic CSC, OCTA in 4 eyes ( 12.50% ) revealed the presence of a distinct choroidal neovascularization (CNV), while no evidence of CNV in ICGA was observed. However, no sign of CNV in acute CSC group both on OCTA and ICGA was found. The occurrence of 'dark areas' in chronic CSC was much higher than that in acute CSC ( P <0.01). In addition, the integrity of the outer retinal structure (defined as tissue between external limiting membrane and retinal pigment epithelium) in acute group was significantly better than that in chronic group ( P <0.01). CONCLU-SION:Our study demonstrates the existing secondary CNV that is not demonstrated by ICGA in the chronic CSC patients, and the different characteristics of retinochoroid structures between acute and chronic CSC in OCTA and SD-OCT are ob- served. Chronic CSC has more severe structural changes.

Objective To develop a BP26 recombinant BCG (rBCG-BP26) vaccine,and to observe the effects of rBCG-BP26 on CD4+,CD8+ T cells in immunized mice.Methods The recombinant shuttle vector pMV261-Ag85B-BP26 was constructed by using traditional molecular biological technology.The recombinant strains were obtained by kanamycin resistance screening and PCR identification after electroporation.Western blotting was used to detect the expression of recombinant BP26 vaccine in immunized mice.Safety experiment was carried out in three different groups:the target experiment(rBCG-BP26) group,the positive control(BCG) group and the negative control(PBS) group,15 BALB/c mice in each group.Intradermal inoculations of 100 μl rBCG-BP26 [containing 106 colony forming units(CFU)],BCG,and PBS were carried out,respectively.Signs of mice in each group were observed.After immunization for 10,20,30,and 40 days,body weight was weighed,and tail blood was collected to observe the change of peripheral blood CD4+ and CD8+ T cells by flow cytometry.Results The rBCG-BP26 was successfully constructed.The expression of BP26 protein was detected in the liquid medium and the bacteria cells.The results of safety test analysis showed that there were no significant differences in signs and body weights(F=2.468,0.331,1.520,0.739,all P＞ 0.05),between PBS group[ (19.24 ± 0.54),(21.37 ± 0.66),(22.83 ± 0.62),(25.06 ± 0.37)g],BCG group[ (19.90 ± 0.02),(21.53 ± 1.57),(21.95 ± 0.55),(24.70 ± 0.39)g]and rBCG-BP26 group[ (19.16 ± 0.55 ),(20.89 ± 0.20),(22.15 ± 0.76),(24.60 ± 0.64)g].The results of flow cytometry showed that the percentages of CD4+ T cell level were lower in BCG group(26.70％,33.07％) and rBCG-BP26 group( 13.40％,26.70％) than that of the PBS group(33.85％,29.33％) and the values of CD4+/CD8+ T cells increased in rBCG-BP26 group (0.69％,1.27％,1.57％,1.70％ ) 10,20 and 30 days after immunization.Conclusions Recombinant BCG-BP26 vaccine strain can express brucella BP26 protein efficiently.Furthermore,its virulence is mild,and it can activate CD4+,CD8+ T cells in the body.It can be used as one of candidate vaccine strain against brucellosis.

Acute Disease ; Case-Control Studies ; Child, Preschool ; Coronavirus NL63, Human ; isolation & purification ; Female ; Humans ; Infant ; Male ; Respiratory Tract Infections ; virology

Objective To investigate the operative effect and value of window partial laryngectomy for the treatment of stage T2-3 glottic laryngeal carcinoma.Methods From Oct.2000 to Dec.2006,the stage T2-3 glottie laryngeal carcinoma of 48 patients by properly selected were performed with window partial laryngectomy and laryngeal reconstruction.Twenty nine males and 19 females were included.Their ages ranged from 43 to 78 years(median 57.0 years).Before operation and in the 6 months after operation,these targets including auditory mental evaluation of hoarse degree,active degree and symmetry of vocal cord,glottie width in the time of quiet breathing,glottie closing degree in the time of phonating,respiratory function and swallowing function were surveyed.In addition,the things of recurrence and metastasis of tumors together with survival time of patients were following investigated. Results All patients were decannulated successfully and incisions were healed smoothly.No operative complication occurred.Except active degree of vocal cord(P=0.343),there were respectively significant difference between two group targets of auditory mental evaluation of hoarse degree(all P＜0.01),symmetry of vocal cord(P=0.000),glottic width in the time of quiet breathing(P=0.001),glottie closing degree in the time of phonating(P=0.001)and respiratory function(P=0.001)those were investigated before operation and after operation.The swallowing function wasn't influeneed(P=0.310).There were laryngostenosis in 1 case,recurrence in 1 case(2.1%),cervical lymph node metastasis in 1 ease and hepatic metastasis in 1 case.Two cases died.3-year and 5-year overall survival rate were respectively 96.9% and 88.9%.Conclusions The study showed that window partial laryngectomy wag successful for treating stage 32-3 glottic laryngeal carcinoma by properly selected.This operation was effective for reducing surgical invasion and beneficial to resume respiratory and vocal function.

OBJECTIVETo investigate the diagnosis and treatment of serious pediatric airway obstruction.

METHODSSeventy three pediatric cases with serious airway obstruction, admitted to Shantou hospital from 1995 to 2005, were analyzed retrospectively.

RESULTSAmong the 73 cases, 28 cases (38.4%) with inflammatory disease and 33 cases (45.2%) with foreign body. In addition, 8 cases of laryngeal papilloma, 3 cases of laryngotracheobronchial spasm and 1 case of pharyngeal dysembryoma were also included in this study. The patients who had endotracheal intubation or tracheotomy were 39 cases (53.4%) and 27 cases (36.9%) respectively. Ten cases received non-surgery treatment. Seventy two cases diagnosis was confirmed and the related serious airway obstruction condition got stable within 12 hours. While serious complications occurred in 23 cases (31.5%) and operative complications occurred in 3 cases (4.1%). Four cases ( 5.5%) died and 68 cases (93.2%) were cured. One case refused to be treated.

CONCLUSIONSThe condition of serious pediatric airway obstruction patients is usually critical, the related causes could be complicated, quick diagnosis and timely removal of the obstruction factors are imperative.

Adolescent ; Airway Obstruction ; diagnosis ; therapy ; Child ; Child, Preschool ; Critical Illness ; Female ; Humans ; Infant ; Male ; Retrospective Studies ; Treatment Outcome

OBJECTIVESTo understand the role cellular immunology plays in the pathogenesis of chronic hepatitis B (CHB) through analysis of T cell receptor (TCR) beta chain variable region gene (BV) family dominant usage and beta chain complementarity determining region3 (CDR3) sequences of peripheral blood mononuclear cells of the patients.

METHODSTCR BV families were amplified by inverse polymerase chain reaction (RT-PCR), and the dominant usage of BV families and CDR3 repertoire were analyzed by immunoscope technology for 8 CHB patients during their acute exacerbations and for 4 healthy blood donors who served as controls. The clonality of the T cells suspected by immunoscope was further confirmed by CDR3 sequencing.

RESULTSThe TCR BV CDR3 repertoire of the 4 healthy blood donors showed a Gaussian distribution. In the 8 CHB patients, however, the clonal expansion of T cells showed different TCR BV families with each patient. The T cells of the clonal expansion shared different CDR3 sequences.

CONCLUSIONThe peripheral blood T cells of CHB patients during their acute exacerbation showed significantly a clonal expansion and their T cell clonal expansion may be stimulated by several HBV epitopes. These results indicate that cellular immunology is involved in the pathogenesis of the liver inflammation process of CHB.

Adult ; Amino Acid Sequence ; Base Sequence ; Cloning, Molecular ; Complementarity Determining Regions ; genetics ; Female ; Gene Rearrangement, beta-Chain T-Cell Antigen Receptor ; genetics ; Hepatitis B, Chronic ; genetics ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Receptors, Antigen, T-Cell ; genetics