BACKGROUND: Effects of Pericarpium aurantiin water extract on sleeping and spontaneous activity in Kunming mice has been investigated in three experiments, that is, the experiments of sleeping time due to supraliminal and subliminal dosage of sodium pentobarbital, and shaking cage for energy transducer, respectively.OBJECTIVE: To observe the influence of Pericarpium aurantiin water extract on sleeping and spontaneous activity of Kunming mice.DESIGN:Randomized controlled trial.SETTING: The Staff Rooms of Pathology, Physiology and Pharmacology in Gannan Medical College.MATERIALS: The experiment was conducted in the Central Laboratory of Science Research, Gannan Medical College from January through February 2005. Ninety adult Kunming mice, grade of clearness, were enrolled and randomly divided into three groups with 30 mice for each experiment:10 in control group, 10 in large-dosage group and 10 in small-dosage group.METHODS: ① In energy transforming trial, spontaneous activity of mice was recorded, small wave with wave amplitude ＜ 5 mm was considered as the waveform of gentle activity in mice, such as fur licking and tickling; middle wave with wave amplitude of 5-10 mm as the waveform of moderate activity, such as walking, standing, climbing up and down;large wave with wave amplitude ＞ 10 mm as the waveform of intensive activity, such as running, jumping up and down. Mice in the small-dosage group, large-dosage group and control group were respectively given intraperitoneal injection of 5 mg/g, 10 mg/g Pericarpium aurantiin water extract and equal volume of normal saline, and placed into the cage after 15 minutes. Then, 3 minutes later, waveforms of mice within 15 s was recorded. ② In the experiments on duration of sleep in mice due to supraliminal or subliminal dosage sodium pentobarbital, 0.02 mL/g or 0.01 mL/g sodium pentobarbital (2.5 g/L) were administrated intraperitoneally into mice in the small-dosage, large-dosage and control group 15 minutes after intraperitoneal injection of 5 mg/g, 10 mg/g Pericarpium aurantiin water extract and equal volume of normal saline were given to the three groups, respectively. Subsequently, comparisons of the number of sleeping mice, time to falling asleep, and sleeping time were carried out among the three groups.MAIN OUTCOME MEASURFS: ① Times of spontaneous activity, ②Sleeping time due to supraliminal dosage of sodium pentobarbital, ③ Duration of sleep due to subliminal dosage of sodium pentobarbital.RESULTS: All the mice were involved in the result analysis without loss.① There were fewer middle and large waves in the small-dosage and largedosage groups as compared with that in the control group (90.5±14.7,58.3±12.5, 45.6±10.1, t=2.341-3.215; P ＜0.05 or P ＜0.01), indicating that Pericarpium aurantiin water extract can evidently inhibit spontaneous activities in mice. ② In comparison with the control group, the time to falling asleep and the duration of sleep due to supraliminal dosage of sodium pentobarbital were shortened and prolonged significantly, respectively,in the small-dosage and large-dosage groups. ③ In comparison with the control group, the time to falling asleep and duration of sleep due to subliminal dosage of sodium pentobarbital were shortened and prolonged significantly, respectively, in the small-dosage and large-dosage groups.CONCLUSION: Pericarpium aurantiin water extract can prolong the duration of sleep and inhibit the spontaneous activities of Kunming mice,which is in cooperation with sodium pentobarbital in central inhibition.

ObjectiveTo investigate the changes in serum soluble complement receptor type 1 (sCR1) at 3 and 6 months after antiviral therapy with telbivudine in patients with chronic hepatitis B (CHB) or liver cirrhosis and the influence of telbivudine on serum sCR1 level. MethodsA total of 57 patients with HBeAg-positive CHB or liver cirrhosis were enrolled and given the antiviral therapy with telbivudine. Venous blood was collected before treatment and at 3 and 6 months after treatment, and enzyme-linked immunosorbent assay was used to measure serum sCR1 level. The paired t-test was used for comparison within one group. ResultsAt 3 and 6 months after the antiviral treatment with telbivudine, the serum sCR1 level changed significantly compared with corresponding baseline values (t=4.864 and 6.238, both P＜0.05). The patients with CHB or liver cirrhosis showed significant changes in the serum sCR1 level at 3 and 6 months after treatment compared with corresponding baseline values (t=3425,5468,4047,7378 all P＜0.05). The patients with CHB had a lower serum sCR1 level at baseline and at 3 and 6 months after treatment than those with liver cirrhosis, but the serum sCR1 level at each time point showed no significant differences between the two groups (all P＞005). ConclusionIn patients with HBeAg-positive CHB or liver cirrhosis, serum sCR1 level is reduced significantly after antiviral therapy with telbivudine.

Objective To observe the clinical effect of monosialotetrahexosyl ganglioside sodium injection (GM1) on spastic cerebral palsy. Methods 98 children with spastic cerebral palsy were randomly divided into control group (n=50) and treatment group (n=48). Both groups received Bobath approach, and the treatment group received GM1 in addition. They were assessed with Functional Independence Measure for Children (WeeFIM), Gross Motor Function Measure (GMFM) and Gesell Development Schedule (GDS) before and after 90 days of treatment. Results The scores of WeeFIM, all the dimensions of GMFM and the gross motor, fine motor, personal-social and adaption of the GDS improved in both groups after treatment (P<0.05), and improved more in the treatment group than in the control group (P< 0.05). Conclusion GM1 may further improve the recovery of function for children with spastic cerebral palsy.

Objective:To observe the occurrence of attention deficit hyperactivity disorder(ADHD) in children aged 3-12 and analyze the related factors of family environment leading to ADHD.Methods:A total of 580 children from June 2020 to December 2021 were selected from the pediatric outpatient clinic of the Children's Hospital of Kunming.According to the diagnostic criteria for children with ADHD in DSM-5, all the subjects were divided into ADHD group ( n=37) and non-ADHD group( n=543). The general data questionnaire and family assessment device (FAD) were used to investigate the relevant data of children.SPSS 25.0 software was used for statistical analysis, and t test, Mann-Whitney U test were used for inter group comparison, and binary Logistic regression model was used to analyze the related factors of family environment of ADHD in children aged 3-12 years. Results:Among 580 children aged 3-12 in pediatric clinic, 37 cases were diagnosed ADHD, accounting for 6.38%.Compared with the non-ADHD group, the breast-feeding time of children in the ADHD group was shorter, the education level of primary caregivers was lower(5.00(2.50, 6.00)months, 9.00(7.00, 11.00)months)( U=7.751), the proportions of families with single parent/reconstituted and parents with criticism and reprimand as the main educational method were higher, and the FAD score((172.35±4.27), (116.29±5.58), t=6.478) was higher (all P<0.05). The results of Logistic regression analysis showed that longer breast-feeding time ( β=-0.561, P<0.001, OR=0.571, 95% CI=0.480~0.678) was the protective factor of ADHD in children aged 3-12 years.Primary and junior high schools as the education level of main caregivers, criticism and reprimand as the main education methods of parents, and poor family function (high FA score) were the related risk factors of family environment of ADHD in children aged 3-12 years. Conclusion:Family environment factors, such as short breast-feeding time, low education level of main caregivers, criticism and reprimand of parents, poor family function, can increase the incidence risk of ADHD in children aged 3-12 years.

OBJECTIVETo investigate the feasibility of reduction in tumor antigen peptide dose by dendritic cell (DC)-presenting so as to elucidate the characteristics of modifying DC by heat shock protein (Hsp70) and antigen peptide.

METHODSAntigen peptide bound to Hsp70 was used to modify DC in vitro. The metabolism of the modified DC and the cytokine secreted thereby was determined. Then the activation of lymphocytes by the modified DC and Hsp70-H22 peptide was tested. The cytotoxicity of the activated lymphocytes to H22 tumor cells and the inhibition of tumor in mice by DC injection and Hsp70-H22 peptide was tested.

RESULTS0.15 micro g of H22 peptide bound to Hsp70 could mature 2 x 10(5) DC. 4 x 10(3) matured DC could activate 2 x 10(6) lymphocytes. The same amount of lymphocyte could be activated to produce similar cytotoxicity to tumor cells by either DC modified by 0.003 micro g of peptides bound with Hsp70 or by direct stimulation with 0.15 micro g of peptides bound to Hsp70. The dose of peptide could be reduced to 1/50 if the modified DC injection was used instead of direct Hsp70-peptide injection. Peptide from the normal hepatocytes, if bound to Hsp70, could not mature DC, nor could it activate lymphocytes through DC.

CONCLUSIONThe dose of Hsp70-H22 peptides can be reduced significantly by DC-presenting to activate lymphocytes. Peptides from normal cells, being unable to activate the lymphocytes by either Hsp70-presenting or DC-presenting, have little to offer in the induction of autoimmunity.

Animals ; Antigens, Neoplasm ; immunology ; Dendritic Cells ; immunology ; Disease Models, Animal ; HSP70 Heat-Shock Proteins ; chemistry ; immunology ; therapeutic use ; Immunity ; Lymphocyte Activation ; Mice ; Mice, Inbred BALB C ; Neoplasm Transplantation ; Neoplasms, Experimental ; prevention & control ; Peptides ; chemistry ; immunology ; therapeutic use ; Tumor Cells, Cultured

By using semi-quantitative RT-PCR method, it was found that PD-L1 mRNA but not PD-L2 mRNA was expressed in H22 hepatoma cells and both PD-L1 and PD-L2 mRNAs were expressed in tumor tissues of tumor-bearing mice and upregulated as compared with muscle tissues in normal mice and H22 hepatoma cells. PD-L1 and PD-L2 were also expressed on the surface of the activated T cells. The soluble recombinant sPD-1 expressed from the constructed eukaryotic expression vector could enhance the lysis of tumor cells by lymphocytes stimulated specifically with antigen. The expresssion of sPD-1 by local gene therapy on the inoculation site of H22 hepatoma cells could inhibit the growth of tumor. The results of this study indicate that expression of soluble receptor of negative costimulatory molecules could reduce the inhibitory effect on T cells in tumor microenvironment and enhance the cytotoxicity of T cells on tumor cells. This possibly provides a new method of improving efficacy of tumor gene therapy.
Animals ; B7-1 Antigen ; biosynthesis ; genetics ; B7-H1 Antigen ; Carcinoma, Hepatocellular ; immunology ; pathology ; Genetic Therapy ; Liver Neoplasms ; immunology ; pathology ; Male ; Membrane Glycoproteins ; biosynthesis ; genetics ; Mice ; Mice, Inbred BALB C ; Peptides ; genetics ; metabolism ; Programmed Cell Death 1 Ligand 2 Protein ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Spleen ; cytology ; T-Lymphocytes ; immunology ; T-Lymphocytes, Cytotoxic ; immunology ; Transfection ; Tumor Cells, Cultured

To investigate the inducement of cytotoxic T lymphocytes (CTLs) by antigen peptides mixture from different leukemia cells and the cross-reaction of the mixtures from different cell lines, antigen peptides mixtures were prepared from different leukemia cell lines respectively and then bound with Hsp70 in vitro. Activation and proliferation of PBMC were observed after stimulation with different Hsp70-peptide complexes. The ratio of CD8+ in proliferative cells was analyzed by flow cytometry. The cytotoxicity of the activated PBMC to different target cells was assayed. The results showed that the antigen peptides from different leukemia cell lines, bound with Hsp70, could activate PBMC effectively, and stimulate the activated PBMC to proliferate. The proliferative PBMC had specific cytotoxicity to corresponding leukemia cells. CD8+ cells, accounting for a high proportion in proliferative cells, had a specific cytotoxicity to leukemia cells from which antigen peptides were prepared, suggesting that these CD8+ cells were CTLs specific to leukemia cells. CTLs activated by Hut78-peptides or Molt4-peptides had a significantly stronger cytotoxicity to Hut78 cells, Molt-1 cells and Jurkat cells than that of CTLs activated by HL-60-peptides (P < 0.05). And the cytotoxicity of CTLs activated by Hut78/Molt4-peptides to Jurkat cells was significantly stronger than that of CTLs activated by either Hut78-peptides or Molt4-peptides alone (P < 0.05). It is concluded that antigen peptides mixtures from leukemia cells can induce specific antitumor CTLs. There exists cross-reactivity among antigen peptides mixtures from different cell lines of the same type leukemia and more cross-reactive antigen peptides could be obtained from more cell lines, suggesting that antigen peptides mixture with broad antigenic spectrum could be prepared by using multiple leukemia cell lines.
Antigens, Neoplasm ; metabolism ; pharmacology ; Cell Division ; Cells, Cultured ; Cross Reactions ; Cytotoxicity, Immunologic ; immunology ; HL-60 Cells ; HSP70 Heat-Shock Proteins ; metabolism ; Humans ; K562 Cells ; Leukocytes, Mononuclear ; cytology ; immunology ; Neoplasm Proteins ; immunology ; Peptides ; pharmacology ; T-Lymphocytes, Cytotoxic ; immunology

Objective:To investigate the value of low-energy virtual monoenergetic image (VMI) at 45 keV in visualizing the primary tumor and T staging of hypopharyngeal squamous cell carcinoma.Methods:The clinical and imaging data of 58 patients with hypopharyngeal squamous cell carcinoma from April 2018 to January 2020 at Eye & ENT Hospital, Fudan University were analyzed retrospectively. All the patients underwent a venous phase contrast-enhanced dual-source dual-energy CT scan before treatment. The VMI at 45 keV and standard linearly blended image (30% 80 kV+70% 140 kV) were acquired from dual-energy post-processing software. One senior radiologist and one junior radiologist independently assessed the visibility of the tumor on the 45 keV VMI and standard linearly blended image using a 5-point Likert rating scale. Furthermore, the senior radiologist assessed the visibility of the tumor at each subsite (piriform fossa, posterior pharyngeal wall, postcricoid region) and determined the invasion depth of the tumor (extension to esophagus, invasion to strip muscles and prevertebral muscles) and performed the T staging of the primary tumor using the two sets of images blindly. The accuracy of T staging was calculated, using pathological T staging (surgical cases) or clinical T staging (non-surgical cases) as the gold standard. The image scores of the two sets of images were compared using Wilcoxon rank sum test. McNemar-Bowker test was used to compare the accuracy of T staging using the two sets of images.Results:The overall image scores of the 45 keV VMI and standard linearly blended image from the senior radiologist were 3.5 (3, 4) and 3 (2, 3) respectively ( Z=-7.03, P<0.001), and the scores from the junior radiologist were 3 (3, 4) and 2 (2, 3) ( Z=-6.93, P<0.001). The scores of the 45 keV VMI were significantly higher than those of the standard linearly blended image in visualizing tumors in the piriform fossa, posterior pharyngeal wall, and postcricoid region, as well as in detecting invasion to the strip muscles ( P<0.05). There was no significant difference in the scores of the two sets of images in determining whether the tumor extended to esophagus or invaded prevertebral muscles ( P>0.05). Referring to pathological and clinical T stage, the accuracy of T staging determined by the 45 keV VMI and standard linearly blended image was 87.9% (51/58) and 81.0% (47/58) respectively, and the difference was not significant (χ 2=3.33, P=0.189). Conclusions:The 45 keV VMI is superior to the standard linearly blended image in visualizing tumors and detecting invasion to the strip muscles of hypopharynx squamous cell carcinoma. However, the accuracy of determining T staging using 45 keV VMI is slightly improved than that of standard linearly blended image, and the difference is not statistically significant. In determining whether the tumor extends to esophagus or invades prevertebral muscles, 45 keV VMI shows no significant advantage over standard linearly blended image.

【Objective】 To analyze the contributing factors of attention deficit hyperactivity disorder (ADHD) in children and the correlation between bone mineral density and physical growth, in order to provide new clues for the prevention and early intervention of ADHD. 【Methods】 A total of 116 children with ADHD were included into the observation group from June 2020 to June 2022, while another 80 healthy children in the same period were included as the control group.Clinical data of the two groups were compared, and the factors influencing ADHD in children were analyzed using multivariate Logistic regression. Children with ADHD were further divided into boys group and girls group based on gender. Bone mineral density (BMD) and physical growth (height and body weight) of the two groups were measured, and the correlation between the two was analyzed. 【Results】 In the observation group, the proportion of boys, maternal bad behaviors during pregnancy, their educational level below high school, critical parenting, and parental stress index were significantly higher compared to the control group (χ²=14.430, 5.689, 5.630, 6.738, t=6.936, P<0.05). Additionally, family environment score was significantly lower than that in the control group (t=6.328, P<0.05). Logistic regression analysis revealed that factors including boys (OR=3.298, 95%CI: 1.759 - 6.184), maternal bad behaviors during pregnancy (OR=2.730, 95%CI: 1.169 - 6.375), maternal education level of senior high school or below (OR=2.032, 95%CI: 1.127 - 3.663), critical parenting (OR=2.349, 95%CI: 1.223 - 4.513), and parental stress index (OR=1.089, 95%CI: 1.055 - 1.124) were positively correlated with ADHD in children (P<0.05), while family environment score was negatively related to ADHD (OR=0.868, 95%CI: 0.820 - 0.919, P<0.05). There were no significant differences in BMD, height and body weight between boys and girls in ADHD group (P>0.05). Pearson correlation analysis indicated a positive correlation of BMD with height and body weight (r=0.409, 0.317, P<0.05). 【Conclusions】 The development of ADHD in children is associated with gender, maternal bad behavior during pregnancy, family parental style and so on. Bone mineral density is closely related to physical growth in children with ADHD, clinical interventions can be implemented to prevent or early intervene ADHD.