Objective To investigate the effect and its mechanism of ziyuglycoside Ⅱ on proliferation, migration, invasion and apoptosis of colon cancer cells HT-29. Methods The effect of ziyuglycoside Ⅱ on cell proliferation of colon cancer cells HT-29 was determined by CCK-8 method; the effect of ziyuglycoside Ⅱ on cell migrative capacity of colon cancer cells HT-29 was determined by scratch assay; the effect of ziyuglycoside Ⅱ on cell invasive capacity of colon cancer cells HT-29 was determined by transwell assay; the effects of ziyuglycoside Ⅱ on cell apoptosis of colon cancer cells HT-29 was determined by flow cytometry; the effects of ziyuglycoside Ⅱ on mRNA and protein expression of protein kinase B (AKT)/phosphatidylinositol-3-kinase (PI3K) signal pathway were determined by real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western-blot, respectively. Results Ziyuglycoside Ⅱ (0, 1, 5, 10, 20, 40, 60 and 80 μmol/mL) inhibited proliferation of colon cancer cells HT-29 in a dose-dependent manner. Ziyuglycoside Ⅱ (5, 10 and 20 μmol/mL) inhibited migration of colon cancer cells HT-29 in a dose-dependent manner. Ziyuglycoside Ⅱ (5, 10 and 20 μmol/mL) inhibited invasion of colon cancer cells HT-29 in a dose-dependent manner. Ziyuglycoside Ⅱ (5, 10 and 20 μmol/mL) promoted apoptosis of colon cancer cells HT-29 in a dose-dependent manner. Ziyuglycoside Ⅱ (5, 10 and 20 μmol/mL) increased mRNA expression of AKT and PI3K, decreased mRNA expression of Caspase-3 and Caspase-9. Ziyuglycoside Ⅱ (5, 10 and 20 μmol/mL) increased protein expression of phosphorylated protein kinase B(p-AKT) and phosphorylphosphatidylinositol-3-kinase (p-PI3K), increased the expression of Cleaved-Caspase-3 and Cleaved-Caspase-9 protein. Conclusion Ziyuglycoside Ⅱ can inhibit proliferation, migration and invasion of colon cancer cells HT-29, and promote the apoptosis of colon cancer cells HT-29. Its mechanism may be related to regulating the signal pathway of AKT/PI3K via promoting phosphorylation of AKT and PI3K protein, activation of Caspase-3 and Caspase-9 protein.

The dental operative microscope has been widely employed in the field of dentistry, particularly in endodontics and operative dentistry, resulting in significant advancements in the effectiveness of root canal therapy, endodontic surgery, and dental restoration. However, the improper use of this microscope continues to be common in clinical settings, primarily due to operators' insufficient understanding and proficiency in both the features and established operating procedures of this equipment. In October 2019, Professor Jingping Liang, Vice Chairman of the Society of Cariology and Endodontology, Chinese Stomatological Association, organized a consensus meeting with Chinese experts in endodontics and operative dentistry. The objective of this meeting was to establish a standard operation procedure for the dental operative microscope. Subsequently, a consensus was reached and officially issued. Over the span of about four years, the content of this consensus has been further developed and improved through practical experience.
Humans ; Dentistry, Operative ; Consensus ; Endodontics ; Root Canal Therapy ; Dental Care

Objective:To investigate the value of the level of heparin-binding protein(HBP)in bronchoalveolar lavage fluid(BALF)on the evaluation of severe pneumonia in children.Methods:A total of 94 children with severe pneumonia who underwent bronchoscopy and bronchoalveolar lavage were admitted at Hunan Children′s Hospital, and HBP levels in BALF were detected.According to the etiological results, the patients were divided into non-bacterial infection group(19 cases) and bacterial infection group(75 cases). According to the existence and severity of acute respiratory distress syndrome (ARDS), the cases were divided into non-ARDS group(65 cases), mild ARDS group(23 cases) and moderate to severe ARDS group(6 cases).Results:The HBP level of BALF in the bacterial infection group was higher than that in the non-bacterial infection group, and the difference was statistically significant[ 20.77(5.90, 73.50)ng/mL vs.5.9(5.90, 7.64)ng/mL, Z=12.500, P<0.001]. The HBP level of BALF in the moderate to severe ARDS group[300.00(169.29, 300.00)ng/mL] was significantly higher than those in the non-ARDS group[11.90(5.90, 36.95)ng/mL] and the mild ARDS group[15.13(7.41, 46.44)ng/mL], and the difference was statistically significant( H=14.718, P=0.001). In predicting the presence of bacterial infection in severe pneumonia, the area under the receiver operating characteristic curves of BALF HBP, serum procalcitonin (PCT) and serum C-reactive protein(CRP) were 0.758, 0.737, and 0.732, respectively.When the optimal truncation values of BALF HBP, serum PCT and serum CRP were 8.40 ng/mL, 0.16 ng/mL, and 8.39 mg/L, the predicted sensitivities were 70.7%, 69.3%, 46.7%, and the predicted specificity were 79.0%, 79.0%, 94.7%, respectively. Conclusion:The level of HBP in BALF in children with severe pneumonia increases with the severity of ARDS, and significantly increases in the positive group of bacterial infection, which can be used as one of the auxiliary indicators to evaluate the severity of severe pneumonia and bacterial infection in children.

Objective:To investigate the etiology, prognosis and clinical characteristics of abnormal serum amylase and lipase in children.Methods:This study was a retrospective study.A total of 7 813 children older than 28 days who had their serum amylase and lipase detected in Hunan Children′s Hospital from August 2017 to August 2020 were included as the study subjects.Children with acute and chronic pancreatitis were excluded.The age, gender, impatient department, imaging exams, discharge outcomes, main diagnosis, diagnostic ICD10 code, and the highest values of serum amylase and lipase during hospitalization were collected through the medical record system.According to the levels of serum amylase and lipase, the children were divided into 3 groups.Patients in group A had normal serum amylase and serum lipase levels.The serum amylase or lipase levels of patients in group B was 1 to 3 times higher than that of group A. The serum amylase or lipase levels in group C was 3 times higher than that of group A. Group B and group C had abnormal pancreatic enzyme levels.According to the prognosis, patients were divided into the survival group and the death group.The relationship of the occurrence of abnormal serum amylase and lipase levels with the age, sex, disease type and prognosis of children was analyzed.Results:The ratio of abnormal trypsin in male and female was 11.5% and 12.9%, respectively.The number of children with abnormal pancreatic enzyme levels in the 28 day -1 year old group, >1-3 years old group, >3-6 years old group, >6 -12 years old group and > 12 year old group were 37 cases (4.6%), 185 cases (15.4%), 199 cases (10.5%), 431 cases (13.9%), and 94 cases (11.7%), respectively.The mortality rate was 1.6% (112/6 867 cases) in group A, 5.2% (32/617 cases) in group B, and 7.6% (25/329 cases) in group C. The mortality risk of group B and C was both higher than that of group A. Compared with group A, the OR (95% CI) of group B and group C was 3.30 (2.21-4.93) and 4.96 (3.17-7.77), respectively.In group C, the top five diseases were parotitis (26.4%), cholangiectasis (11.6%), choledochal cysts (8.5%), gastroenteritis (4.5%) and sepsis (3.3%). Conclusions:Pancreatic enzyme abnormalities in children are associated with adverse prognosis.Pancreatic enzyme abnormalities are more prone to occur in children aged >1-3 with mumps, digestive diseases and congenital digestive system structural deformities.In addition, children with sepsis are also easy to present pancreatic enzyme abnormalities.Clinical attention should be paid to the possibility of secondary pancreatic damage in children with sepsis.

Objective:To investigate the expression of Macrophage migration-inhibitory factors (MIF) in hepatocellular carcinoma (HCC) tissues and its interaction with ERK1/2 signaling pathway, so as to establish a theoretical basis for further studying the molecular mechanism of MIF promoting HCC.Methods:From February 2020 to August 2021, 52 cases of hepatocellular carcinoma (HCC) tissues based on hepatitis B cirrhosis (HBV-LC) and 52 cases of adjacent tissues in Tianjin Medical University Cancer Hospital and 940th Hospital of Joint Logistic Support Force of PLA were collected as the experimental group, including 39 males and 13 females, aged 35-65 years. And 20 cases of normal liver tissue were selected as the control group. Immunohistochemistry was used to detect the expression of MIF, ERK1/2 and p-ERK1/2 proteins in liver tissues of the two groups, and in situ hybridization was used to detect the expression of ERK1/2 nucleic acid in liver tissues of the two groups.HepG2 HCC cells and L-02 normal hepatocytes were co-cultured with different concentrations of rMIF, the expression and phosphorylation levels of ERK1/2 and JNK1 proteins in the two kinds of liver cells were detected by Western-blot, and the expression levels of ERK1/2 nucleic acids in the two kinds of liver cells were detected by RT-PCR. One-way ANOVA was used for measurement data and χ 2 test was used for counting data. Results:The expressions of MIF, ERK1/2, p-ERK1/2 and ERK1/2 mRNA were significantly increased in HCC and para-cancer tissues (the expression of MIF in HCC group was 78.8%, and that in adjacent group was 75.0%; ERK1/2 80.8% in HCC group and ERK1/2 71.8% in paracancerous group. The expression of p-ERK1/2 75.0 % in HCC group and 46.2% in paracancerous group were respectively detected. ERK1/2 mRNA was expressed in HCC group 76.9%, ERK1/2 mRNA expression in paracancerous group 78.8%), and the differences were statistically significant compared with normal liver tissues ( P<0.05), but there was no significant difference between HCC and para-cancer tissues ( P>0.05). The expressions of ERK1/2, p-ERK1/2 and ERK1/2 mRNA in HepG2 HCC cells were significantly increased with the increase of rMIF concentration, and the increase was most obvious when rMIF concentration was 200 ng/ml, and the difference was statistically significant compared with L-02 normal hepatocytes ( P<0.05). Conclusion:MIF, ERK1/2 and p-ERK1/2 are highly expressed in HCC tissues and HepG2 HCC cells, suggesting that MIF promotes the occurrence and development of hepatocellular carcinoma through ERK1/2 signaling pathway.

Objective:To evaluate the effect of serum cytokines and immune typing on the severity and prognosis of children with sepsis.Methods:One hundred and sixty children with sepsis admitted to No.2 PICU from May 2018 to May 2019 at Hunan Children′s Hospital were enrolled.Vital signs, blood routine and blood biochemical indexes of the children were collected within 24 hours after admission.Serum cytokine concentrations of interleukin(IL)-2, IL-4, IL-6, IL-10, tumor necrosis factor(TNF)-α, and interferon (IFN)-γ were tested at the same time.According to the worst condition during the first 24 h after admission, and the consensus diagnostic criteria of experts in the diagnosis and treatment of septic shock in children in 2015, the patients were divided into sepsis group(76 cases) and severe sepsis group(84 cases). According to the prognosis of 28 days, the patients were divided into survival group(134 cases)and death group(26 cases). The differences of cytokine concentration and the composition of immune state among groups were compared.Results:The levels of IL-2[2.40(2.40, 2.55)pg/mL vs. 2.40(2.40, 3.76)pg/mL], IL-6[60.54(23.22, 250.38)pg/mL vs. 21.47(8.83, 57.94)pg/mL], IL-10[50.85(21.74, 217.12)pg/mL vs. 14.99(9.23, 28.79)pg/mL] , TNF-α[2.75(2.40, 5.15)pg/mL vs. 2.40(2.40, 3.23)pg/mL] and IFN-γ[8.01(4.34, 37.28)pg/mL vs.2.62(2.62, 28.56)pg/mL] in the severe sepsis group were significantly higher than those in the sepsis group( P<0.05). The levels of IL-10[99.57(10.51, 646.96)pg/mL vs. 23.17(11.74, 57.30)pg/mL] and TNF-α[3.08(2.40, 5.13)pg/mL vs. 2.45(2.40, 3.80)pg/mL] in the death group were significantly higher than those in the survival group( P<0.05). The risk of death in children with mixed antagonistic response syndrome (MARS) was significantly higher than those without MARS( OR 3.75, 95% CI 1.302-10.804). Conclusion:The increased concentrations of IL-2, IL-6, IL-10, TNF-α, and IFN-γ are significantly correlated with the severity of sepsis in children.Increased IL-10 concentration and the occurrence of MARS are significantly correlated with poor prognosis.The occurrence of MARS in children with sepsis should be highly vigilant for the poor prognosis.

Objective:To investigate the changes of inflammatory factors, serum amylase and lipase and pancreatic damage at different time points after establishment of cecal ligation and perforation in a rat model of sepsis, which provides a stable and reliable animal model for the study of pancreatic injury following sepsis in children.Methods:Sixty male SD rats were randomly divided into 6 h group, 12 h group, 24 h group and 48 h group in control group, sham group and cecal ligation and puncture (CLP) group.Rat model of sepsis was established by cecal ligation and perforation in CLP group, the sham group was treated with sham operation, and rats in control group were not treated.The levels of interleukin (IL)-1β, IL-10, amylase and lipase in each group were detected, and pancreatic tissue was taken for HE staining.Results:In this model, the 48 h survival rate of rats in the CLP group was 70%.With the prolongation of postoperative time in the CLP group, pancreatic tissue congestion, cell edema, and inflammatory cell infiltration gradually worsened.The pathological changes of pancreatic tissue in the Sham group were mild.The expression levels of IL-1β in CLP group and sham group increased first and then decreased, and reached the peak at 12 h. The expression levels of IL-1β in CLP group were significantly higher than those in control group and sham group at all time points( P<0.05). The expression level of IL-10 showed a gradually increasing trend, reaching a peak at 48 h. The expression level of IL-10 in CLP group was significantly higher than that in control group and sham group at all time points( P<0.05), which in sham group was significantly higher than that in control group( P<0.05). Serum amylase and lipase in CLP group showed an upward and then downward trend, reaching the peak at 24 h. The serum amylase levels at 24 h and 48 h were significantly higher than those in control group and sham group( P<0.05). Serum amylase levels in sham group were significantly different from those in control group at 12 h, 24 h and 48 h( P<0.05). Serum lipase level at 12 h, 24 h and 48 h was significantly higher than that in control group and sham group( P<0.05). There was no significant difference in serum lipase between sham group and control group( P>0.05). Conclusion:After CLP modeling, IL-1β, IL-10 elevation and pancreatic pathological changes appeared at the early stage, and the pathological changes gradually increased with time.Significant differences in serum amylase and lipase began to appear at 24 h and 12 h respectively.

Objective:To summary the mixed infection as well as clinical characteristics and analyze the risk factors for mixed infection of severe adenovirus pneumonia(SAP) in children.Methods:The clinical data of 114 children with SAP were retrospectively analyzed.Multivariate Logistic regression analysis was performed to assess the risk factors for mixed infection.Results:The incidence age was from 6 months to 2 years(62.5%). High fever(94.7%), cough(98.2%), dyspnea(86.8%) and lethargy(95.6%) were the main symptoms.Laboratory examination showed that children with SAP were prone to increased white blood cell count, C-reactive protein, procalcitonin, aspartate aminotransferase, alanine aminotransferase and CK-MB, as well as decreased proportion of CD3 + , CD4 + , CD8 + , CD4 + /CD8 + and NK cells.The main complications intrapulmonary organ were respiratory failure(80.7%). The main complications extrapulmonary organ were circulatory complications (55.3%). SAP was easily combined with other pathogenic infections.Streptococcus pneumoniae(22.9%)was the most common bacterial pathogen.Respiratory syncytial virus(10.0%)were the most common virus, in addition, mycoplasma pneumoniae(17.1%) was also common.Multivariable Logistic regression analysis showed that the decreasing ratio of CD4 + /CD8 + and NK cells, congenital heart disease and congenital airway dysplasia were the independent risk factors for mixed infection of SAP in children( P<0.05). Conclusion:The SAP patients could easily suffer from mixed infection and high fatality rate.Immune dysregulation is the important risk factors for mixed infection of SAP in children.So immunoregulatory treatment is very important.

Objective:To analyze the epidemiological characteristics, clinical characteristics and blood purification effect of mushroom poisoning in children.Methods:A retrospective study was conducted on 51 children with acute mushroom poisoning admitted to Hunan Children′s Hospital from 2002 to 2020.The epidemiological and clinical characteristics were analyzed, and the prognosis of children with different incubation periods was analyzed and compared.Among them, 36 critically ill children were treated with blood purification.Results:The age distribution was 66(43, 115)months.Mushroom poisoning had obvious seasonal and spatial aggregation.The first symptom was mainly manifested by digestive tract, which was manifested as vomiting, abdominal pain and diarrhea, accounting for 94.1% patients(48/51). The gastrointestinal tract type accounted for 45.1%(23/51) of the clinical types and the multi-organ damage type accounted for 51.0% patients(26/51). The length of hospital stay was 6(3, 11)days.Among them, early onset accounted for 45.1% patients(23/51), late onset accounted for 54.9% patients(28/51). Early onset hospitalization was shorter, about 4(2, 7)days, and fewer organs 1(0, 3) were damaged.The length of hospital stay of late onset was 8(3, 12)days, and the number of damaged organs was 4(2, 4). There was a statistically significant difference( P<0.05). Late onset patients had more severe organ function damage, including liver function, coagulation function, renal function, myocardial enzyme, and there was no significant difference in the final outcome between two groups.In 36 children who were given blood purification treatment, alanine aminotransferase, aspartate aminotransferase, prothrombin time, international normalized ratio, blood urea nitrogen, and lactate dehydrogenase were significantly improved, with statistically significant differences( P<0.05). Conclusion:Mushroom poisoning has obvious seasonal and geographical distribution characteristics; the first manifestation is mainly gastrointestinal symptoms; the clinical types are more common in gastrointestinal and multiple organ damage types.Children with early onset have shorter hospital stays and fewer complications than later onset.Blood purification treatment can significantly improve liver function, kidney function and blood coagulation function in children with toadstool poisoning.

Objective:To discuss the role of continuous blood purification (CBP) therapy in children with severe adenovirus pneumonia.Methods:A total of 114 children with severe adenovirus pneumonia admitted to the Department of PICU at Children′s Hospital of Hunan Province from June 2018 to July 2019 were selected as the research objects.According to whether treated with CBP, they were divided into CBP group and control group.The following indicators during the process of treatment were compared between two groups, including respiratory mechanics indicators[respiratory index(PaO 2/FiO 2), dynamic lung compliance(Cdyn)]; hemodynamic indicators(heart rate and mean arterial pressure); changes in levels of inflammatory factors interleukin(IL)-6, IL-10, tumor necrosis factor(TNF)-α and the prognosis 28 days after admission. Results:The respiratory mechanics index, serum IL-6 and TNF-α levels of two groups after treatment were significantly lower than those before treatment, and the serum IL-10 level was significantly higher than that of this group before treatment.There were statistical differences in the CBP group before and after treatment, while there was no statistical difference in control group.In the CBP group, the serum IL-6 and TNF-α levels after treatment were significantly lower than those of the control group( P<0.05), and the serum IL-10 level was significantly higher than that of the control group( P<0.05). The 28-day mortality rate of patients in CBP group was 8.6%(3/35), which was significantly lower than 13.9%(11/79) of control group ( P<0.05). Conclusion:CBP could improve the main respiratory mechanical indexes of adenovirus pneumonia and decrease the level of inflammatory cytokines.