AIM:To investigate the changes of autophagy of macrophages after phagocytizing dust particles and explore the mechanisms of dust particle-induced autophagy of the cells.METHODS:The bronchopulmonary lymph nodes were collected from patients with lung operation.The paraffin sections were prepared and then stained with Wilder's method.The tissue structures were viewed.Peritoneal macrophages were harvested from rats and then treated with carbon particles.Influences of carbon particles in autophagic activities of macrophages were examined.The ultrathin sections of the lymph nodes and the cells phagocytized carbon particles were prepared.The structures and distribution of phagosomes,autophagosomes and lysosomes were viewed.The apoptotic cells in the dust cells of the lymph nodes and the cells having phagocytized carbon particles were examined using transmission electron microscope and TUNEL staining.RESULTS:In adult lymph nodes,dust particles were deposited significantly in macrophages,collagen fibres and density of microvessels increased.There were autophagosome precursors,autophagosomes,autophagolysosomes as well as phagosomes in the dust cells and the cells phagocytized carbon particles.In autophagosomes,mitochondrion,dust particle or carbon particle were usually observed.There were positive cells by TUNEL staining in the dust cells and the cells phagocytized carbon particles.Nuclear condensation or apoptotic body in the apoptotic cells were observed under transmission electron microscope.CONCLUSION:Deposition of dust particles induces enhancement of autophagic activities and apoptosis of macrophages.Autophagy plays an important role in cleaning dust particles and the injured mitochondria.

Objective To investigate the effects of antisense recombinant euraryotic expression vector of HCCR-2 on the proliferation and apoptosis of HepG2. Methods The antisense recombinant eukaryotic expression vector of HCCR-2 was constructed. The vector was stably transfected to the HepG2 cells, and positive clones were selected by G418 (antiseuse vector group), pIRES2-EGFP vector was transfected into the HepG2 cells in the same way (pIRES2-EGFP group). The conditions of the nontransfected HepG2 cells were used as control (HepG2 group). Changes in cell growth curve, cell cycle, cell apoptosis and morphology of HepG2 cells after the transfec-tion were detected by MTT method, flow cytometry and transmission electron microscopy, respectively. All the data were analyzed by one-way ANOVA and chi-square test. Results The expression level of HCCR-2 mRNA was down-regulated to 0.39±0.04 in antisense vector group, and the expression level of HCCR-2 mRNA in pIRES2-EGFP group and HepG2 group were 0.62±0.06 and 0.72±0.03, respectively, with significant difference among the 3 groups (F=43.701, P<0.05). The apoptotic rate of HepG2 cells in antisense vector group, pIRES2-EGFP grop and HepG2 group were 13.30%, 2.51% and 2.07%, respectively, with significant difference among the 3 group (χ2=6.793, 8.721, P<0.05). The growth of HepG2 cells in antisense vector group was retarded, and was blocked in G0/G1 stage. Conclusions The HCCR-2 antisense recombinant eukaryotic expression vector can inhibit the mRNA expression of HCCR-2 and promote the apoptosis of cells. HCCR-2 may be involved in cell regulation and the proliferation of hepatocellular carcinoma cells.

OBJECTIVE:To establish a method for the simultaneous determination of 5 unsaturated fatty acids in Perilla oil cap-sule. METHODS:With the reference material of α-linolenic acid methyl ester,GC was used to determine and calculate the relative correction factors of α-linolenic acid methyl ester with methyl palmitate,methyl stearate,methyl oleate and linoleic acid methyl es-ter,and the correction factors were used to calculate the contents of 5 unsaturated fatty acids;the column was Agilent Innowax cap-illary column,the detector was FID,the inlet temperature was 230 ℃,the detector temperature was 250 ℃,the gas flow rate was 20 ml/min(nitrogen),40 ml/min(hydrogen)and 350 ml/min(air),split ratio was 30 to 1,the column temperature was 190 ℃, and injection volume was 1 μl. RESULTS:The linear range was 0.018-0.792 μg(r=0.9994)for methyl palmitate,0.0016-0.0176μg(r=0.9993)for methyl stearate,0.0056-0.2464 μg(r=0.9999)for methyl oleate,0.003-0.132 μg(r=0.9990)for linoleic acid methyl ester and 0.018-0.792 μg(r=0.9998) for α-linolenic acid methyl ester;RSDs of precision,stability and reproducibility tests were lower than 5%;recoveries were 98.990%-101.70%(RSD=0.720%,n=6) for methyl palmitate,99.599%-100.699%(RSD=0.368%,n=6) for methyl stearate,98.996%-101.680%(RSD=1.240%,n=6) for methyl oleate,99.813%-100.963%(RSD=0.434%,n=6)for linoleic acid methyl ester and 97.185%-99.602%(RSD=0.874%,n=6)for α-linolenic acid methyl es-ter. CONCLUSIONS:The method is simple and stable with good reproducibility,and can be used for the simultaneous determina-tion of methyl palmitate,methyl stearate,methyl oleate,linoleic acid methyl ester,α-linolenic acid methyl ester in Perilla oil cap-sule.

OBJECTIVE: This study was designed to explore the risk factors associated with severity of juvenile onset recurrent respiratory papillomatosis. METHOD: A retrospective study was conducted to study determinants of severe forms of juvenile recurrent onset respiratory papillomatosis. The patients were separated into different groups based on the onset age, the first recurrence of age, the first recurrence of period, gender and incision of tracheal respectively. The relationship among the lesion severity score,the involvement of the subregion, operation period and the next operation period were also explored. RESULT: It was observed that some children who recurred before 4 years old required more surgery, shorter operation period(the average, longest or shortest operation period) than those elder children, the differences were statistically (P=0. 029, 0. 003, 0. 010, 0. 039, respectively). The severity score of lesion was correlated positively with the involvement of the subregion and negatively with operation period (r=0. 914, -0. 451, respectively). Some children who diagnosed before 4 years old had to endure more severity score and shorter operation period than those older children, the differences were statistically (P= 0. 036, 0. 000, respectively). 8 cases accepted incision of tracheal, they accepted more surgery too. But the differences in the onset age, the first recurrence of age, and the operation period were not statistically. CONCLUSION The results showed that the clinical course of juvenile onset recurrent respiratory papillomatosis was closely related to the first recurrence age and period, while the severity of disease was associated to the onset age and the involvement of the subregion.
Adolescent ; Age of Onset ; Child ; Child, Preschool ; Humans ; Papilloma ; Papillomavirus Infections ; classification ; epidemiology ; surgery ; Respiratory Tract Infections ; classification ; epidemiology ; surgery ; Retrospective Studies ; Risk Factors ; Severity of Illness Index ; Trachea

OBJECTIVE: To explore the relationship between serum IL-4, IFN-gamma, IL-10 levels and the aetiology of juvenile-onset recurrent respiratory papillomatosis. METHOD: Serum IL-4, IFN-gamma, IL-32 levels of 15 JORRP children were detected by use of enzyme-linked immunosorbent assay (ELISA) and compared with those of healthy control group. RESULT: Serum IL-4 levels were significantly higher in the JORRP children (P<0.01): (524.65 +/- 147.77)pg/ml in the JORRP children and (213.27 +/- 87.48) pg/ml in the healthy control group. Serum IFN-gamma levels were significantly lower in the JORRP children (P<0.01): (2.87 +/- 0.84) pg/ml in the JORRP children and (10.63 +/- 5.09) pg/ml in the healthy control group. Serum IL-32 levels were significantly lower in the JORRP children (P< 0.01): (2.47 +/- 1.60) pg/ml in the JORRP children and (9.08 +/- 2.66) pg/ml in the healthy control group. CONCLUSION 1) While the concentration of Th2 like cytokine IL-4 in children with JORRP was higher than that in control group, the concentration of Th1 like cytokine IFN-gamma in children with JORRP was lower than that in controls, indicating that the polarization of Th1 /Th2 T cell in children with JORRP; 2) The polarization of Th1/Th2 T cell may cause the reduction of the serum IL-32 as a proinflammatory role in host immunity system that could not eradicate HPVs because of lacking enough inflammatory stimulation.
Case-Control Studies ; Child ; Female ; Humans ; Infant ; Interferon-gamma ; blood ; Interleukin-4 ; blood ; Interleukins ; blood ; Male ; Papillomavirus Infections ; blood ; Respiratory Tract Infections ; blood

BACKGROUND/AIMS: Gastrointestinal (GI) symptoms may develop when we fail to adapt to various stressors of our daily life. Central oxytocin (OXT) can counteract the biological actions of corticotropin-releasing factor (CRF), and in turn attenuates stress responses. Administration (intracerebroventricular) of OXT significantly antagonized the inhibitory effects of chronic complicated stress (CCS) on GI dysmotility in rats. However, intracerebroventricular administration is an invasive pathway. Intranasal administration can rapidly deliver peptides to the brain avoiding stress response. The effects of intranasal OXT on hypothalamus-pituitary-adrenal axis and GI motility in CCS conditions have not been investigated. METHODS: A CCS rat model was set up, OXT 5, 10, or 20 μg were intranasal administered, 30 minutes prior to stress loading. Central CRF and OXT expression levels were analyzed, serum corticosterone and OXT concentrations were measured, and gastric and colonic motor functions were evaluated by gastric emptying, fecal pellet output, and motility recording system. RESULTS: Rats in CCS condition showed significantly increased CRF expression and corticosterone concentration, which resulted in delayed gastric emptying and increased fecal pellet output, attenuated gastric motility and enhanced colonic motility were also recorded. OXT 10 μg or 20 μg significantly reduced CRF mRNA expression and the corticosterone concentration, OXT 20 μg also helped to restore GI motor dysfunction induced by CCS. CONCLUSION: Intranasal administration of OXT has an anxiolytic effect and attenuates the hypothalamus-pituitary-adrenal axis in response to CCS, and gave effects which helped to restore GI dysmotility, and might be a new approach for the treatment of stress-induced GI motility disorders.
Administration, Intranasal ; Animals ; Anti-Anxiety Agents ; Brain ; Colon ; Corticosterone ; Corticotropin-Releasing Hormone ; Gastric Emptying ; Gastrointestinal Motility ; Models, Animal ; Oxytocin ; Peptides ; Rats ; RNA, Messenger

OBJECTIVE: To explore the clinical and genetic characteristics of a patient with 17-hydroxylase/17,20-lyase deficiency. METHODS: The patient was infertile without contraception. Laboratory examination showed her chromosomal karyotype to be 46, XX. DNA sequencing was performed to detect variants of CYP17A1 gene in the patient and her family members. RESULTS: Sanger sequencing revealed that the patient has carried homozygous variant c.1486C>T in the exon 8 of the CYP17A1 gene, which resulted in substitution of arginine by cysteine (p.Arg496Cys). Her family members were all heterozygotes for the same variant. CONCLUSION Homozygous variant of the CYP17A1 gene c.1486C>T probably underlay the 17-hydroxylase deficiency in this patient. Above finding has enabled accurate genetic counseling and prenatal diagnosis for her family.

Objective:To investigate the effect of different stent oversize in thoracic endovascular aortic repair (TEVAR) on lumen remodeling of type B aortic dissection (TBAD).Methods:The clinical and follow-up data of 89 TBAD patients receiving TEVAR from Nov 2010 to Jun 2020 at Yantai Yuhuangding Hospital were retrospectively analyzed. According to the difference of proximal stent oversize, 89 patients were divided into: low oversize group (<10%, 47 cases) and high oversize group (≥10%, 42 cases). The changes of the normal vessel diameter and area at the proximal end of the stent and the long diameter, short diameter and area of the true/false lumen at the distal end of the stent at 3, 6, and 12 months after surgery and postoperative complications were analyzed.Results:The change of proximal vessel diameter with time in the low oversize group is smaller than that in the high oversize group ( P<0.05),and the change of the distal false lumen area of the stent in the low oversize group was greater than that in the high oversize group ( P<0.05). The high oversize group was prone to retrograde type A aortic dissection (RTAD) ( P<0.05). Conclusion:Low oversize stents are more conducive to the remodeling of the aortic lumen in the early and mid-term after TEVAR in TBAD patients.

OBJECTIVETo investigate the correlation between nonsyndromic deafness and mitochondrial 12s rRNA A839G mutation.

METHODSAccording to the clinical manifestations of mitochondrial DNA sequencing and analysis to find and determine family containing mitochondrial 12s rRNA A839G mutation. Harvested its family members blood and transferred their lymphocytes into lymphoblastoid cell lines, followed by cells cultured, cell doubling experiment, susceptibility testing, cellular oxygen consumption rate experiment, ROS and mitochondrial membrane potential experimental tests were progressed to explore the correlation between the A839G mutation and nonsyndromic deafness.

RESULTSThe mitochondrial 12s rRNA A839G mutation pedigrees were determined through the full sequence detections of the Mitochondrial DNA, further phylogenetic analysis showed that 839 point conservative index (CI) up to 78.6%; in RPMI-galactose medium containing A839G gene mutant cell line, the doubling time was significantly longer than the control group, and the difference was significant (P = 0.033). The effect to cell lines containing the A839G mutation of aminoglycoside drugs was not obvious. When compared with the control group, cell lines containing the A839G mutation significantly reduced cellular oxygen consumption rate(P = 0.033); compared with the control group, the ROS levels of cell lines containing the A839G mutation appeared more substantial elevated with significan difference (P < 0.01). The mitochondrial membrane potential of cells of experimental group was significantly reduced than the control group.

CONCLUSIONThe present study proved that the mitochondria 12s rRNA A839G mutations affect the function of the mitochondrial respiratory chain at the cell level, which might reduce the growth rate of the mutant cell lines, result in hearing.

Aminoglycosides ; Cell Line ; DNA, Mitochondrial ; Deafness ; genetics ; Galactose ; Hearing Tests ; Mitochondria ; Mutation ; Pedigree ; Phylogeny ; RNA, Ribosomal ; genetics

Spindle assembly checkpoint (SAC) is a protective mechanism for cells to undergo accurate mitosis. SAC prevented chromosome segregation when kinetochores were not, or incorrectly attached to microtubules in the anaphase of mitosis, thus avoiding aneuploid chromosomes in daughter cells. Aneuploidy and altered expression of SAC component proteins are common in different cancers, including lung cancer. Therefore, SAC is a potential new target for lung cancer therapy. Five small molecule inhibitors of monopolar spindle 1 (MPS1), an upstream component protein of SAC, have entered clinical trials. This article introduces the biological functions of SAC, summarizes the abnormal expression of SAC component proteins in various cancers and the research progress of MPS1 inhibitors, and expects to provide a reference for the future development of lung cancer therapeutic strategies targeting SAC components.
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Humans ; Cell Cycle Proteins/metabolism* ; Spindle Apparatus/metabolism* ; Protein Serine-Threonine Kinases/metabolism* ; M Phase Cell Cycle Checkpoints/genetics* ; Lung Neoplasms/metabolism*