Objective We used the DNA methylation microarrays to investigate the differential methylation genes and loci sites in Kashin-Beck disease (KBD),to study the relationship between DNA methylation and KBD pathogenesis.Methods Totally 12 KBD adults and 12 healthy adults were selected and peripheral blood samples were collected and DNA was extracted.Illumina 450K bead-chip was applied to detect methylation status in KBD and healthy controls.Aberrant hyper-methylated sites were filtrated according to the P value after correction and methylation differences,together with GenomeStudio soft.Screened genes were validated using bisulfite sequencing polymerase chain reaction (BSP) technology.Results A total of 484 948 loci sites were analyzed and compared,93 differential methylated loci were found by comparing KBD and normal people,including 34 hypermethylated sites and 59 hypomethylated sites.There were 50 genes corresponding to the loci,43 genes not reported in literature.According to gene ontology analysis,the genes were involved in the immune response,antigen processing,phosphate and phosphoric acid metabolism and phosphorylation and the process of metal ions in combination.However,in the verification test using BSP method,there was no significant difference in methylation rate in human leukocyte antigen (HLA)-DRB1 between the case and the control group (48％ vs 70％,x2 =3.688,P ＞ 0.05).Conclusions The high and low differentially methylated sites in peripheral blood DNA of KBD patients are significantly different from those of the health control.HLA-DRB1 locus is not significantly different between the BSP verification test and methylation chip.

T-2 toxin is a secondary fungal metabolite that belongs to the trichothecene mycotoxin family.T-2 toxin can lead to the structural and functional changes of cartilage cells and cartilage cell degeneration and necrosis.With strong cytotoxicity,T-2 toxin can cause definite damage to cartilage cells.In this paper,we reviewed recent studies on the effects of T-2 toxin on chondrocyte apoptosis,ultra structural changes and extracellular matrix in vitro.

Aim To explore micro RNAs-integrated pathogenic signaling to control endothelial-mesenchy-mal transition ( EndMT ) in pulmonary hypertension ( PH) by a network bioinformatic approach. Methods Literature-mining method was used to find PH-relat-ed genes and EndMT/EMT-related miRNAs. Bioinfor-matic prediction approach ( DIANA3 , Miranda4 , PicT-ar5 , TargetScan6 , miRDB7 and microT-CDS8 ) was used for miRNA target prediction. Hypergeometric a-nalysis was used to predict miRNAs related to EndMT in PH. The analysis of interactions between PH-rele-vant genes( PH network) was performed with the use of Biological General Repository for Interaction Datasets ( BioGRID ) . These miRNAs were ranked with the highest probability of substantial overlap among their gene targets in the PH-network, the relationship be-tween their targets and the PH functional categories which include hypoxia, inflammation, and transforming growth factor/BMP signaling. Then, the part of results was validated by animal experiment. Lastly the miR-NA-Target network was built using Cytocape 3 . Results List of 230 genes was compiled that were directly im-plicated in the development of PH and 189 miRNAs were related to EndMT in PH. Among 189 miRNAs, only 22 microRNAs(miR-let-7 family, miR-124, miR-130 family, miR-135, miR-144, miR-149, miR-155, miR-16-1, miR-17, miR-181 family, miR-182, miR-200 family, miR-204, miR-205, miR-21, miR-224, miR-27, miR-29 family, miR-301a, miR-31, miR-361 and miR-375) were related to hypoxia, inflamma-tion, and transforming growth factor/BMP signaling. Among these miRNAs, the levels of let-7g, miR-21, miR-124 and miR-130 family were significantly changed in the pulmonary artery in hypoxia-induced PH rats. Conclusions Among numerous miRNAs,22 of which may be involved in hypoxia, inflammation, and transforming growth factor/BMP signaling and re-lated to EndMT in PH by network bioinformatic ap-proach, which provides a theoretical basis for further investigation of EndMT in PH.

Objective:The paper aims at developing a method of defining and visualizing the scope of the basic medical insurance pharmacy service, and provides a new way of thinking for the designated pharmacy planning. Methods:Collecting the basic data and information on administrative divisions in the planning area taking equity and efficiency as the guidance, using ArcGIS and its function modules to define and visualize the scope of the medical in-surance pharmacy service. The procedure of issue focus, method improvement, data simulation, expert consultation, methodology perfecting were followed to define and visualize the scope. Results:Forming a whole set of operative pro-cedures of defining and visualizing the scope of the medical insurance pharmacy service based on medical resources allocation standard, and the operation commands and procedures in ArcGIS were clarified. Conclusion:Operating ac-cording to the appropriate method steps, the following can be achieved:(1) The adjacent scope of medical insurance pharmacy service are adjacent to each other but do not overlap or cross;(2) Spatial relations can be clearly and ef-fectively expressed;(3) The shape is flat and regular;(4) The data collected at different times can be comparable in space, providing good prerequisites for medical insurance designated pharmacy planning.

Objective:To systematically review the association between delivery mode and exclusive breastfeeding rate during hospitalization and within the first six months of life.Methods:Observational studies on the association between delivery mode and feeding pattern were searched from PubMed, Web of Science, Cochrane Library, EBSCO, China Biomedical Literature Database, CNKI, Wanfang Database, and VIP Database from inception to October 2022. Two independent reviewers screened the literature, extracted data, and assessed the quality of included studies using Critical Appraisal Tools published by Joanna Briggs Institute or Newcastle-Ottawa Quality Scale (NOS). This meta-analysis was performed using R 4.1.0 software. Fixed-effect or random-effect models were used to pool data. Egger test and funnel plot were used to assess publication bias.Results:A total of 34 studies involving 597 203 subjects were included, including 22 cross-sectional studies and 12 cohort studies. All of the 22 cross-sectional studies were B-level quality, and eleven out of the 12 included cohort studies scored 7 points or above on the NOS scale with high quality. The results of meta-analysis showed that the likelihood of exclusive breastfeeding during hospitalization of women who had cesarean section was lower than those who delivered vaginally ( OR=0.33, 95% CI: 0.22-0.50, P<0.001); and so was the likelihood of exclusive breastfeeding at six months postpartum ( OR=0.61, 95% CI: 0.47-0.79, P<0.001). Conclusion:Current evidence suggests that cesarean section is a disadvantage to exclusive breastfeeding during hospitalization and within six months after delivery.

BACKGROUND:Due to the treatment of cervical spondylosis,the Zero-P system of the anterior cervical interbody fusion system will have problems such as screw loosening and fracture after operation,so a novel Low-P system has been developed. OBJECTIVE:To compare the effects of the novel Low-P and Zero-P anterior cervical intervertebral fusion systems on the biomechanical properties of adjacent segments of the cervical spine and to perform stress analysis on the internal fixation system,so as to provide a theoretical reference for clinical treatment. METHODS:A complete model of the C1-C7 segment of the cervical spine was established.Based on the effectiveness of the model,a finite element model of Low-P(type Z Low-P and type H Low-P)and Zero-P system implanted in C4-C5 segments was established.The stress distribution of implanted devices and adjacent vertebral nucleus pulposus,fibrous rings and end plates was analyzed under the conditions of forward flexion,posterior extension,lateral bending and rotation. RESULTS AND CONCLUSION:(1)After implantation of Low-P and Zero-P internal fixation devices,the range of motion of the type H Low-P system was large;the maximum stress value of type Z Low-P system was small;the maximum stress of Zero-P on the nucleus pulposus of adjacent segments was large;the maximum stress of end plate was small.(2)The influence of three internal fixation systems on adjacent segment fiber rings was close.(3)The screw stress of the Zero-P internal fixation system was much greater than that of the Low-P system.(4)It is indicated that compared with Zero-P type internal fixation system,the novel Low-P system reduces the stress value of steel plate and screw,which can reduce screw loosening and internal fixation system failure.The Low-P system has less stress on the nucleus pulposus of adjacent discs and reduces disc degeneration in adjacent segments.This paper provides a theoretical basis for the clinical study of a Low-P type internal fixation system.

OBJECTIVE: To explore the clinical features, lysosomal enzymatic [acid α-glucosidase (GAA)] activities and genetic variants in a child with late-onset Pompe disease (LOPD). METHODS: Clinical data of a child who had presented at the Genetic Counseling Clinic of West China Second University Hospital in August 2020 was retrospectively analyzed. Blood samples were collected from the patient and her parents for the isolation of leukocytes and lymphocytes as well as DNA extraction. The activity of lysosomal enzyme GAA in leukocytes and lymphocytes was analyzed with or without addition of inhibitor of GAA isozyme. Potential variants in genes associated with neuromuscular disorders were analyzed, in addition with conservation of the variant sites and protein structure. The remaining samples from 20 individuals undergoing peripheral blood lymphocyte chromosomal karyotyping were mixed and used as the normal reference for the enzymatic activities. RESULTS: The child, a 9-year-old female, had featured delayed language and motor development from 2 years and 11 months. Physical examination revealed unstable walking, difficulty in going upstairs and obvious scoliosis. Her serum creatine kinase was significantly increased, along with abnormal electromyography, whilst no abnormality was found by cardiac ultrasound. Genetic testing revealed that she has harbored compound heterozygous variants of the GAA gene, namely c.1996dupG (p.A666Gfs*71) (maternal) and c.701C>T (p.T234M) (paternal). Based on the guidelines from the American College of Medical Genetics and Genomics, the c.1996dupG (p.A666Gfs*71) was rated as pathogenic (PVS1+PM2_Supporting+PM3), whilst the c.701C>T (p.T234M) was rated as likely pathogenic (PM1+PM2_Supporting+PM3+PM5+PP3). The GAA in the leukocytes from the patient, her father and mother were respectively 76.1%, 91.3% and 95.6% of the normal value without the inhibitor, and 70.8%, 112.9% and 128.2% of the normal value with the inhibitor, whilst the activity of GAA in their leukocytes had decreased by 6 ~ 9 times after adding the inhibitor. GAA in lymphocytes of the patient, her father and mother were 68.3%, 59.0% and 59.5% of the normal value without the inhibitor, and 41.0%, 89.5% and 57.7% of the normal value with the inhibitor, the activity of GAA in lymphocytes has decreased by 2 ~ 5 times after adding the inhibitor. CONCLUSION The child was diagnosed with LOPD due to the c.1996dupG and c.701C>T compound heterozygous variants of the GAA gene. The residual activity of GAA among LOPD patients can range widely and the changes may be atypical. The diagnosis of LOPD should not be based solely on the results of enzymatic activity but combined clinical manifestation, genetic testing and measurement of enzymatic activity.
Humans ; Child ; Male ; Female ; Glycogen Storage Disease Type II/pathology* ; Retrospective Studies ; alpha-Glucosidases/genetics* ; Mothers ; Lysosomes/pathology* ; Mutation

Objective To study the effects of different doses of T-2 toxin on organs and bones of BALB/c pregnant mice and their offspring mice.Methods Sixty female SPF BALB/c mice at the age of 6-weeks were mated with 30 6-week-old male SPF BALB/c mice.Female mice were divided into control,low dose,medium dose and high dose groups according to body weight via the random digital table method,with 15 mice in each group.After mating with male mice,the pregnant mice in each group were 14,10,9 and 15,respectively.Nutritional interventions (feed:the doses of T-2 toxin were 0,600,1 200 and 2 400 ng/g,respectively) were initiated from the gestation day 0 until the first generation mice were grown up (6-weeks-old).The growth status and organ coefficient (heart,liver,kidney,thymus,spleen and brain) of the two generations in each period were recorded.Skeletal X-ray photographs of the two generations were taken by digital radiography.The histopathological changes in the organs (liver,thymus,spleen and epiphyseal cartilage) of the two generations were observed under light microscope.Results Among the pregnant mice,there were no significant differences in organ coefficients (P ＞ 0.05).No abnormalities were observed in each group of skeletal X-ray photographs.In the female generation mice,there were no significant differences in the coefficients of heart,liver and kidney (F =0.233,2.196,0.430,P ＞ 0.05),while there were significant differences in the coefficients of thymus,spleen and brain (F=3.683,3.148,4.498,P ＜ 0.05),and the thymus coefficient of medium dose group was higher than that of control group;the thymus coefficient of high dose group was lower than that of other three groups;the spleen coefficients of the three dose groups were higher than that of control group;the brain coefficients of the three dose groups were lower than that of control group (P ＜ 0.05).In the male generation mice,there were no significant differences in the coefficients of thymus and brain (F =2.447,1.620,P ＞ 0.05),while there were significant differences in the coefficients of heart,liver,kidney and spleen (F =5.339,2.738,11.435,2.872,P ＜ 0.05),and the heart coefficient of high dose group was lower than that of control group and low dose group;the coefficients of liver and kidney in medium dose and high dose groups were lower than those in control and low dose groups;the spleen coefficients of the three dose groups were higher than that of control group (P ＜ 0.05).The pathological changes of liver,thymus,spleen and epiphyseal cartilage were found in dose exposure groups;epiphyseal hyperplasia was found in the skeletal X-ray photographs of medium and high dose groups.Conclusion T-2 toxin has no significant effects on pregnant mice,but it could cause damage to the organs and epiphyseal plate cartilage of the first generation,and the location of the injury is related to gender.

Objective To investigate the effects of different doses of T-2 toxin on the expression of cytokines cytokines and pathological changes in parental mice and their offspring. Methods One hundred female mice and 25 male mice (CD-1, SPF) were adapted for one week. After regular random mating, observation of vaginal suppository within the first 24 hours was as the 0th day of pregnancy. The pregnant rats were divided into high dose, medium dose, low dose and control groups according to body weight by a random number table(Feed: the doses of T-2 toxin were 1 200, 600, 300, and 0 μg/kg, respectively), with 16 - 18 rats in each group. The high, middle and low dose groups began to consume the poisoned feed on the 0th day of pregnancy, while the control group consumed the standard feed. After natural delivery, their offspring were continually treated the same way as their mother until the offspring reached adulthood. Serum levels of interleukin-1β (IL-1β) and interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), organ coefficient and pathological changes of articular cartilage were determined. Results The levels of IL-1β,IL-6 and TNF-α in the control, low, middle and high T-2 toxin groups during the pro-pregnancy of the middle-aged mice were [(219.56 ± 19.32), (136.89 ± 20.41), (210.49 ± 21.23), (207.41 ± 21.23); (192.73 ± 22.43), (136.25 ± 29.55), (187.43 ± 39.32), (232.48 ± 39.32); (1 303.02 ± 142.10), (1 072.60 ± 78.30), (1 065.03 ± 37.44), and (1 169.72 ± 104.18) ng/L], respectively. The differences between control and T-2 toxin treated groups were statistically significant (F = 17.124, 6.237, 7.670, P < 0.05). For further pairwise comparison,IL-1β and IL-6 in low dose group were significantly lower than those in control, middle and high dose groups (P < 0.05); TNF-α content in control group was significantly higher than those in low,middle and high dose groups(P<0.05).There were significant differences in the levels of IL-1β,IL-6 and TNF-α between the control group and the low,middle and high dose groups of offspring weanling mice[(142.36 ± 13.36),(113.01 ± 8.65), (102.13 ± 8.31), (123.42 ± 10.41); (109.92 ± 9.76), (100.26 ± 15.60), (85.25 ± 9.97), (100.21 ± 16.46);(1 308.45 ± 204.90), (1 248.60 ± 96.85), (1 081.09 ± 105.51), (1 204.87 ± 153.96) ng/L, F = 49.823, 10.530, 7.490, P < 0.05]. The levels of IL-1β and IL-6 in the control group were significantly higher than those in the low, middle and high dose groups(P < 0.05).The levels of TNF-α in the control group were significantly higher than those in the medium and high dose groups(P < 0.05).The levels of the three cytokines IL-1β, IL-6 and TNF-α in adult filial mice were significantly different [(69.71 ± 9.61), (61.31 ± 10.07), (63.07 ± 10.39), (58.56 ± 9.69); (172.55 ± 24.55),(146.91 ± 13.47),(151.02 ± 24.93), (157.21 ± 17.86); (1 136.87 ± 137.39), (1 002.22 ± 86.52), (987.12 ± 130.80),(1 047.21 ± 171.64)ng/L, F=4.670,5.636, 4.775, P < 0.05], the contents of the three cytokines in the poisoning groups were significantly lower than that of the control group (P < 0.05). The organ coefficients of thymus, spleen and liver in the second trimester were significantly different [(0.14 ± 0.03), (0.20 ± 0.06), (0.15 ± 0.02), (0.12 ± 0.03); (0.71 ± 0.16), (0.78 ± 0.14), (0.77 ± 0.15), (0.38 ± 0.10); (6.19 ± 0.43), (5.57 ± 0.57), (6.04 ± 0.32), (5.11 ± 0.29), F = 4.056, 11.064, 8.312, P < 0.05], and the thymus index was significantly increased in low dose group (P<0.05),spleen coefficient decreased significantly in high dose group (P < 0.05), and liver coefficients in low and high dose group were significantly decreased (P < 0.05). In the offspring, the midbrain coefficient of viscera showed significant changes [(3.45 ± 0.73), (3.11 ± 0.31), (2.98 ± 0.45), (3.04 ± 0.22), F = 7.529, P < 0.05], which was significantly decreased in the exposed rats(P<0.05).Both the mid-pregnant mice and filial mice showed varying degrees of changes in epiphyseal cartilage injury. The degree of epiphyseal cartilage injury became higher with increasing dosages of T-2 toxin in mid-pregnancy and post-weaning parental mice, and the injury was more serious in post-weaning mice. Conclusions Exposure to T-2 toxin can cause decrease of cytokines IL-1β, IL-6 and TNF-α in the blood of CD-1 pregnant and filial mice, and also cause the cartilage damage in mice, which are aggravated following increased doses of T-2 toxin and extension of exposure time.

Kaschin-Beck disease is an endemic and deformed chronic osteochondropathy. Though the etiology is not well clear, the etiologic hypotheses have been mainly focused on bio-geochemical hypotheses, the hypotheses of mycotoxin poisoning under low selenium condition and the hypotheses of toxic organic compounds in drinking water. Prevention and control measures based on these hypotheses have shown remarkable achievements. Depending on the related research at home and abroad, this paper reviews the new developments of pathogen, pathogenesis and prevention on Kaschin-Beck disease in recent years.In terms of scientific research,new progresses have been made in the aspects of environmental factors associated with the pathogenesis of Kaschin-Beck disease, molecular biology, genomics, proteomics and environmental response genes. As for prevention and treatment, new progress has been made in such fields as supplement of selenium and treatment of traditional Chinese medicine.