Objective In this study,we analyzed the transcriptome sequences of Kupffer cells exposed to simulated microgravity for 3 d and conducted biological experiments to determine how microgravity initiates apoptosis in Kupffer cells. Methods Rotary cell culture system was used to construct a simulated microgravity model.GO and KEGG analyses were conducted using the DAVID database.GSEA was performed using the R language.The STRING database was used to conduct PPI analysis.qPCR was used to measure the IL1B,TNFA,CASP3,CASP9,and BCL2L11 mRNA expressions.Western Blotting was performed to detect the level of proteins CASP3 and CASP 9.Flow cytometry was used to detect apoptosis and mitochondrial membrane cells.Transmission electron microscopy was used to detect changes in the ultrastructure of Kupffer cells. Results Transcriptome Sequencing indicated that simulated microgravity affected apoptosis and the inflammatory state of Kupffer cells.Simulated microgravity improved the CASP3,CASP9,and BCL2L11 expressions in Kupffer cells.Annexin-V/PI and JC-1 assays showed that simulated microgravity promoted apoptosis in Kupffer cells.Simulated microgravity causes M1 polarization in Kupffer cells. Conclusion Our study found that simulated microgravity facilitated the apoptosis of Kupffer cells through the mitochondrial pathway and activated Kupffer cells into M1 polarization,which can secrete TNFA to promote apoptosis.

OBJECTIVES: To provide reference basis for reducing the mortality for children under 5 years old and promote the healthy development, the mortality for children under 5 years old and the main causes for death in Liuyang City from 2013 to 2020 are analyzed. METHODS: The data of 725 cases of death for children under 5 years old in Liuyang City from 2013 to 2020 were collected.The causes and difference of death among the children were analyzed retrospectively by descriptive statistic methods. RESULTS: There were a total of 144 516 live births in Liuyang City from 2013 to 2020. The mortality for children under 5 years old was 5.01‰, for infants was 3.39‰, and for newborns was 1.63‰. The male child mortality was 5.28‰, and the female child mortality rate was 4.72‰, with significant difference (P>0.05). The mortality for children under 5 years old was seasonal fluctuation, without significant difference among seasons (P>0.05). For the past 5 years, the top 3 causes for death among children under 5 years old were preterm birth and low birth weight, congenital heart disease, and pneumonia. Before death, 341 cases (47.04%) were treated in provincial hospitals, 198 cases (27.31%) in county-level hospitals, 56 cases (7.72%) in village-level hospitals, and 130 cases (17.93%) were not treated. CONCLUSIONS The mortality for children under 5 years old in Liuyang City is gradually reduced in the past 5 years. The main causes for death are premature birth and low birth weight, congenital heart disease and pneumonia. We should develop healthy education, improve the rate of prenatal diagnosis, promote the construction of obstetrics and paediatrics, and fundamentally reduce the mortality for children under 5 years old.
Cause of Death ; Child ; Child Mortality ; Child, Preschool ; Female ; Heart Defects, Congenital ; Humans ; Infant ; Infant Mortality ; Infant, Newborn ; Male ; Pneumonia/epidemiology* ; Pregnancy ; Premature Birth ; Retrospective Studies

ObjectiveTo obtain HSC-T6 cells with stable expression of Cas9 protein and HSC-T6-COX-2-/- cells with COX-2 gene defect by transfecting HSC-T6 cells with CRISPR/Cas9 lentiviral vector, and to provide a good method for further functional research and new strategies for the clinical treatment of liver fibrosis. MethodsThe COX-2 gene-specific sgRNAs (COX-2-sgRNA-1, COX-2-sgRNA-2, COX-2-sgRNA-3) were designed, synthesized, and connected to the GV371 vector, and the recombinant plasmid and the packaging plasmid were transfected into 293T cells to form lentivirus particles; the fluorescence method was used to measure virus titer. The most appropriate amount of the virus was calculated based on MOI. Lenti-Cas9-puro was transfected into HSC-T6 cells, and HSC-T6-Cas9 cells were screened out by puromycin; Lenti-COX-2-sgRNA-EGFP was transfected into HSC-T6-Cas9 cells to obtain HSC-T6-COX-2-/- cells. Cruiser enzyme digestion and Western blot were used to verify gene knockout at the gene and protein levels. An analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t-test was used for further comparison between two groups. ResultsSequencing verified that the COX-2-sgRNA expression vector was constructed successfully. Recombinant expression plasmids and packaging plasmids were transfected into 293T cells to form lentivirus particles, and the fluorescence method showed a virus titer of ＞1×108. HSC-T6 cells with stable expression of Cas9 protein and HSC-T6-COX-2-/- cells with COX-2 gene defect were successfully constructed. The HSC-T6-Cas9 group had significantly higher relative mRNA expression of LV-Cas9-Puro than the CON group (541.93±105.76 vs 1.00±0.02, t=12.995, P＜0.01). Cruiser enzyme digestion and Western blot showed that the CRISPR/Cas9 lentivirus expression vectors played a role in the target, among which COX-2-sgRNA-2 knockout had the most significant effect, and this group had a significant reduction in the protein expression level of COX-2 compared with the CON group and the NC group (both P＜0.05), suggesting that COX-2-sgRNA was active. ConclusionA CRISPR/Cas9 lentivirus vector is successfully constructed for COX-2 target gene, and HSC-T6-COX-2-/- cells with stable COX-2 gene knockout are obtained.

ObjectiveTo investigate the effect and significance of cyclooxygenase-2 (COX-2) inhibitors on the expression of the Acsl gene family in the ileum of rats with nonalcoholic fatty liver disease (NAFLD). MethodsA total of 45 Sprague-Dawley rats were randomly divided into normal control group (15 rats given normal diet), NAFLD model group (15 rats given high-fat diet), and nimesulide group (15 rats given high-fat diet and nimesulide). All rats were sacrificed after 12 weeks of feeding, and then blood samples were collected from the inferior vena cava to measure total cholesterol (TC) and triglyceride (TG). HE staining and oil red O staining were performed for the liver to evaluate the degree of hepatic steatosis in each group, and quantitative real-time PCR was used to measure the mRNA expression of the Acsl family genes in the ileum. An analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the normal control group, the NAFLD model group had significant increases in serum TC and TG and marked hepatic steatosis (all P＜0.05); compared with the NAFLD model group, the nimesulide group had significant reductions in serum TC and TG and degree of hepatic steatosis (all P＜0.05). Compared with the normal control group, the NAFLD model group had a significant increase in the expression of COX-2 in the ileum (P＜0.05), and compared with the NAFLD model group, the nimesulide group had a significant reduction in the expression of COX-2 in the ileum (P＜005). Compared with the normal control group, the NAFLD model group had significant increases in the mRNA expression of Acsl3 and Acsl5 in the ileum (both P＜0.05), and compared with the NAFLD model group, the nimesulide group had significant reductions in the mRNA expression of Acsl3 and Acsl5 (both P＜0.05). ConclusionThe COX-2 inhibitor nimesulide can regulate the expression of the Acsl gene family in the ileum of rats with NAFLD, suggesting that COX-2 inhibitors may inhibit the progression of NAFLD through the Acsl gene.

Objective:To analyze the malignant probability of thyroid nodules with the diagnosis of atypia of undetermined significance or follicular lesion of undetermined significance (AUS/FLUS) determined by fine-needle aspiration (FNA) and to explore the value of the combined application of BRAFV600E gene detection for the diagnosis of benign and malignant thyroid nodules. Methods:A total of 114 patients including 20 males and 94 females, aged 16-76 years old with thyroid nodules underwent FNA examination and surgical treatment in the Affiliated Cancer Hospital of Zhengzhou University from October 2018 to November 2019 were retrospectively analyzed. Postoperative histopathological results were used as the gold standard for the diagnosis of malignant thyroid nodules. The malignant rate of thyroid nodules with the diagnosis of AUS/FLUS was evaluated. Differential diagnostic efficacy of preoperative FNA combined with BRAFV600E gene detection for papillary thyroid carcinoma (PTC) was analyzed by McNemer test and diagnostic test evaluation method. Results:The mutation rate of BRAFV600E gene was 84.76% (89/105) in PTC. PTC accounted for 57.14% (12/21) of the patients with the diagnoses of AUS/FLUS determined by FNA. The specificity, sensitivity, positive predictive value and negative predictive value of BRAFV600E mutation examination for the diagnosis of malignant thyroid nodules determined preoperatively as AUS/FLUS were 9/9, 5/12, 5/5 and 9/16, respectively. BRAFV600E mutation examination could improve the detection rate of PTC in patients with AUS/FLUS ( OR=0.438, 95% CI=0.251-0.763, P=0.016). Conclusion:FNA combined with BRAFV600E mutation examination can significantly improve the detection rate of malignant thyroid nodules diagnosed preoperatively as AUS/FLUS.

Objective: To investigate the histological and biomechanical changes of rabbit ear cartilage induced by long pulse width 1064 nm Nd: YAGlaser. Methods: Seven New Zealand rabbits weighing 2.0-2.5 kg, male, 4-6 months old were selected. For a self-control, the right ear was irradiated by laser and the left ear was the normal control group. Each rabbit ear was divided into 3 cm × 1 cm, three 2 cm × 1 cm and 1 cm× 1 cm experimental area. After the experimental area of the laser irradiation group was irradiated by long pulse width 1064 nm Nd: YAG laser, Three pieces of 2 cm × 1 cm cartilage in each ear of the experimental rabbits were immediately cut and stained with HE, Sirius red and Masson to observe the histological changes of chondrocytes. The 3 cm × 1 cm and 1 cm× 1 cm cartilage of each rabbit ear was cut for biomechanical examination, and the changes of biomechanical properties such as tension, fracture, elastic modulus and compression of rabbit ear cartilage were observed immediately after the operation. Results: Histological observation showed that the chondrocytes became small, the matrix was stained deeply, the refraction was obvious/evident and the collagen was relatively increased. After being irradiated by long pulse width 1064 nm Nd: YAG laser, the peak tensile stress, tensile elastic modulus, tensile fracture load, compressive stress peak, compressive elastic modulus and compressive failure load of rabbit ear cartilage in laser irradiation group were (5.22 ±0.80) MPa, (42.40 ±9.78) MPa, (22.86 ±4.85) N, (16.04 ±5.57) MPa, (28.71 ±13.97) MPa, (1 211.63 ±427.86) N. All of them were smaller than those of the control group[(6.07±0.64) MPa, (48.44±6.30) MPa, (26.94±4.19) N, (25.12±9.10) MPa, (45.30±19.24) MPa, (1 962.83±896.71) N], and the difference was statistically significant (P<0.05). Conclusions After the operation of long pulse width 1064 nm Nd: YAG laser, the stress of rabbit ear cartilage resisting elastic deformation is reduced, the shape is easier to change, and the remodeling effect is more stable.

Objective To investigate the relationship between serum uric acid level or hyperuricemia of healthy adults and gender , age, ethnicity and altitude in plateau area .Methods 17 358 healthy adults were randomly selected from Qinghai People′s Hospital Physical Examination Center from January 2016 to December 2016.We detected serum uric acid of these healthy controls and analyzed its relationship between gender , age and ethnicity.Besides, serum uric acid data of 7 596 healthy adults were collected to investigate its relationship between altitude .Those adults who were from different altitude areas were localized in Tibet.The serum uric acid was detected by Roche Modular DPP automatic biochemical analyzer.Results The serum uric acid of healthy adults who lived in Qinghai at 2 300 m above sea level generally formed a normal distribution.Serum uric acid of male and female was (371.14 ±69.33)μmol/L and (271.38 ±54.49) μmol/L respectively.Multivariate linear regression analysis showed that gender and ethnicity were the main factors which influence serum uric acid level at the same altitude (2 300 m).The serum uric acid levels and high uric acid detection rates between different genders and ethnicity at the same altitude were statistically significant .The serum uric acid levels and high uric acid detection rates of hyperuricemia among Tibetan healthy adult males at different altitudes were statistically significant .The simulated equation based on the elevation of serum uric acid and altitude difference was Y＝0.041X+4.474 (Y:the rise of serum uric acid, X:Bad altitude,R2＝0.716).Conclusions Gender, ethnicity and altitude are the main factors influencing serum uric acid level and the high rate of hyperuricemia of healthy adult in Qinghai Plateau, among which, that of Tibetan are significantly higher than other ethnic groups .And serum uric acid levels increase along with altitude.

Objective To investigate the application of noninvasive correction of congenital auricle defor-mity,and to analyze the cure rate and the causes of incidence of complications. Methods From January 2015 to December 2016,33 patients with unilateral or bilateral congenital auricle deformities were treated with non-surgi-cal correction technique in the Plastic Surgery Department of Guangzhou General Hospital. When the treatment was completed,the VAS score was used to assess the satisfaction of the patient's parents. Results From January 2015 to December 2016,33 patients(53 ears)received noninvasive treatment of the ear deformity correction system. The average age of the patients was(63.5 ± 41.3)days,the average treatment time was(41.6 ± 15.4)days,and the average parental satisfaction was 3.76 ± 0.93(total score:5 points). Significant difference was observedin auri-cle morphology data between pre- and post- treatment ,except for cranioauricular angle(P < 0.05). Conclusions In this study,the cure rate for non-surgical correction was good,however,the satisfaction of treatment decreased with age.The most common complication of treatment was auricle skin damage. The reasons of causing complica-tions maybe the age-induced reduction of ear cartilage elasticity which brought great pressure on the ear ,and the increase of the treatment time.

Objective To prepare a kind of Gelsolin-targeted ultrasound contrast agent (GSN-PLGA) and to explore its targeting and imaging effection in vitro.Methods The high molecular PLGA-COOH ultrasound contrast agents were prepared by a modified double emulsion technique and then conjugated with Gelsolin monoclonal antibody by carbodiimide technique.The physical property of contrast agent was determined.And the connectivity condition of ultrasound contrast agent with Gelsolin monoclonal antibody was estimated.The targeting ability and the effect of enhancing ultrasound imaging in vitro were explored.Results The average diameter of GSN-PLGA was (575.67 ± 4.71) nm.The potential was (-11.46±1.19) mV.And the binding rate of Gelsolin monoclonal antibody was 96.93％.In vitro experimentshowed more GSN-PLGA could be intaked by Hca-F cells and the ultrasound imaging cloud be enhanced greatly.Conclusion The GSN-PLGA nanoparticle can bind to Hca-F cells specifically and can enhance the ultrasound imaging greatly.

Objective To apply quality control circle (QCC) to reducing non-timely maintenance rate of medical equipment.Methods QCC was established,and some measures were taken to observe its effect in decreasing non-timely maintenance rate of medical equipment.Results The non-timely maintenance rate fell from 12.1％ to 4.5％,and 0.3 million Yuan was saved for the manpower cost.Conclusion QCC decreases the non-timely maintenance rate of medical equipment,increases the efficiency of the maintainer,shortens the downtime of the fault medical equipment,enhances hospital economic benefit and patient satisfaction and provides references for other hospitals.