Objective:To explore the effects of NOD2 gene on the proliferation and apoptosis of tongue squamous cell carcinoma Tca8113 cells.Methods:NOD2 expression vector(NOD2-pEZ-M29)and NOD2-shRNA vector were established,then were trans-fected into Tca8113 cells respectively.Expressions of HBD-2 and NOD2 in the cells was detected by RT-PCR and Western blot.Cell proliferation was examined by MTT assay and apoptosis by flow cytometry at 48h post transfection.Results:Compared with the control group,the expression of NOD2 and HBD-2 in NOD2-pEZ-M29 transfection group was significantly higher and markedly lower in NOD2-shRNA group.The proliferation rate of Tca8113 cells was markedly lower in NOD2-pEZ-M29 transfection group and signifi-cantly higher in NOD2-shRNA group while the apoptosis rate was significantly higher in NOD2-pEZ-M29 transfection group and sig-nificantly lower in NOD2-shRNA group.Conclusion:In Tca8113 cells NOD2 expression was positively correlated with HBD-2 ex-pression.NOD2 gene may promote the apoptosis,inhibit the proliferation of Tca8113 cells.

Objective To compare the mesothelin expressions in 3 human pancreatic cancer cell lines between in vitro and in vivo and the developing speed among the subcutaneous tumors implanted with the 3 human pancreatic cancer cell lines in nude mice.Methods The human pancreatic cancer cell lines ( SW1990,BxPC3 and PANC1 ) were cultured and then were implanted subcutaneously into left axillas of nude mice.The volumes of these subcutaneous tumors were recorded every week to estimate their developing speed.The mice implanted with SW1990 and BxPC3 cells were observed for three weeks,while the mice implanted with PANC1 cell were observed for five weeks.The Western blot method was used to measure the expressions of mesothelin in the 3 kinds of cells and subcutaneous tumors,while immunohistochemical staining was applied to determine the expressions of mesothelin in 3 kinds of subcutaneous tumors.Results The sequence of quantities of expressions of mesothelin in these cell lines in vitro were BxPC3 ＞ PANC1 ＞ SW1990,and the sequence of quantities of expressions in vivo were SW1990 ＞ BxPC3 ＞ PANC1.One handrued percent of the tumors grew out successfully,and the sequence of speeds of their growth was SW1990 ＞ BxPC3 ＞ PANC1.Conclusions The mesothelin expressions among 3 kinds of pancreatic cancer cell line are different.The developing speeds of tumors originated from different subcutaneous tumors in nude mice are also different,and there is no association between them.

OBJECTIVE: Genetic screening and prenatal diagnosis was performed in eighteen families with high risk of 21-hydroxylase deficiency (21-OHD) to provide valuable information for genetic counseling in these affected families. METHODS: First, multiplex ligation-dependent probe amplification (MLPA) combined with nested-PCR based Sanger sequencing was used to detect CYP21A2 gene mutations in probands and their parents of eighteen families, with seven probands had been dead. Second, paternity test was applied to exclude the possibility of maternal genomic DNA contamination, and fetal prenatal diagnosis is based on the mutations found in proband or parents of the family. RESULTS: Ten mutations were identified in these eighteen families, including large fragment deletion, I2G, E3del8bp, I172N, V281L, E6 cluster, L307Ffs, Q318X, R356W and R484Pfs. All probands were caused by homozygous or compound heterozygous mutations of CYP21A2 gene and their parents were carriers. By comparing short tandem repeat sites contamination of maternal genomic DNA was not found in fetal DNA. Prenatal diagnosis showed that five fetus were 21-OHD patients, four fetus were carriers and the other nine fetus were normal. CONCLUSION CYP21A2 gene mutation is the etiology of 21-OHD. Genetic testing of CYP21A2 could assist physicians in 21-OHD diagnosis and provided genetic counseling and prenatal diagnosis for parents who are at risk for having a child with congenital adrenal hyperplasia.
Adrenal Hyperplasia, Congenital ; diagnosis ; genetics ; Female ; Genetic Testing ; Humans ; Mutation ; Pregnancy ; Prenatal Diagnosis ; Steroid 21-Hydroxylase

OBJECTIVETo detect potential mutation of iduronate-2-sulfatase (IDS) gene in a family affected with mucopolysaccharidosis type Ⅱ (MPS Ⅱ).

METHODSFor the proband and his unaffected mother, the whole coding sequence of the IDS gene was analyzed with PCR and bidirectional Sanger sequencing.

RESULTSA novel splicing mutation, c.709-1G>A, was detected in the proband, for which his mother was heterozygous.

CONCLUSIONThe c.709-1G>A splicing mutation of the IDS gene is probably causative for the MSP Ⅱ in the proband. Prenatal diagnosis for the mutation may avoid birth of further child affected with this disease.

Base Sequence ; Child ; DNA Mutational Analysis ; methods ; Family Health ; Female ; Genetic Predisposition to Disease ; genetics ; Glycoproteins ; genetics ; metabolism ; Heterozygote ; Humans ; Iduronate Sulfatase ; genetics ; metabolism ; Male ; Mothers ; Mucopolysaccharidosis II ; diagnosis ; enzymology ; genetics ; Mutation

OBJECTIVE: To explore the genetic etiology for a pedigree affected with progressive familial intrahepatic cholestasis (PFIC). METHODS: Target sequence capture and next generation sequencing (NGS) were applied for the proband. PCR and Sanger sequencing were used to verify the suspected mutation in his sister with similar symptoms and his parents. RESULTS: The proband and his sister manifested after birth with symptoms including jaundice, pruritus and developmental retardation. NGS has identified compound heterozygous mutations of ABCB11 gene, which encodes bile salt export pump protein (BSEP), namely c.2494C>T (p.Arg832Cys) and c.3223C>T (p.Gln1075*), in the proband, which were inherited from his father and mother respectively. His sister carried the same compound mutations. CONCLUSION Based on the phenotype and genetic testing, the patients were diagnosed as PFIC2 caused by mutation of the ABCB11 gene. The c.3223C>T is a novel nonsense mutation which may cause premature termination of translation. Above results have enriched the spectrum of ABCB11 mutations and provided new evidence for the molecular basis of PFIC, which also facilitated genetic counseling for this pedigree.
ATP Binding Cassette Transporter, Subfamily B, Member 11 ; genetics ; ATP-Binding Cassette Transporters ; Cholestasis, Intrahepatic ; genetics ; Female ; Genetic Testing ; Humans ; Male ; Mutation ; Pedigree ; Phenotype

Regulating the catalytic activity of nanozymes is significant for their applications in various fields.Here,we demonstrate a new strategy to achieve reversible regulation of the nanozyme's activity for sensing purpose.This strategy involves the use of zero-dimensional M0S2 quantum dots(MQDs)as the building blocks of nanozymes which display very weak peroxidase(POD)-like activity.Interestingly,such POD-like activity of the MQDs largely enhances in the presence of Fe3+while diminishes with the addition of captopril thereafter.Further investigations identify the mechanism of Fe3+-mediated aggregation-induced enhancement of the POD-like activity and the inhibitory effect of captopril on the enhance-ment,which is highly dependent on their concentrations.Based on this finding,a colorimetric method for the detection of captopril is developed.This sensing approach exhibits the merits of simplicity,rapidness,reliability,and low cost,which has been successfully applied in quality control of captopril in pharmaceutical products.Moreover,the present sensing platform allows smartphone read-out,which has promising applications in point-of-care testing devices for clinical diagnosis and drug analysis.

Tongue diagnosis is an important method of TCM diagnosis and treatment.Tongue is the key link of auxiliary diagnosis of tongue feature extraction and processing,and also is the bottleneck of intelligent tongue diagnosis in traditional Chinese medicine.Using image processing,artificial intelligence technology to the tongue as a quantitative and identify characteristics of traditional Chinese medicine,looking for both conforms to the original thinking of TCM,and TCM tongue diagnosis method of accurately,has become a common concern of traditional Chinese medicine and computer field.From the mobile terminal tongue as auxiliary diagnostic system of traditional Chinese medicine tongue acquisition basic attribute,tongue diagnosis and image information building,tongue like features are required for accurate extraction and so on related key technology is analyzed,and build overall architecture,so as to provide technical reference for the tongue like intelligent diagnosis,promote the development of technology of tongue diagnosis in traditional Chinese medicine modernization.

Objective:To investigate the effects and mechanism of metformin on the wound healing of full-thickness skin defects in diabetic rats.Methods:This study was an experimental study. Eighteen 8-week-old male Sprague Dawley rats were divided into control group, diabetes group, and diabetes+metformin group according to complete random grouping method, with 6 rats in each group. The latter two groups of rats were used to create diabetic models, and then four circular full-thickness skin defect wounds with a diameter of 5 mm were made on the back of 18 rats. Metformin F-127 hydrogel was applied only to the wounds of rats in diabetes+metformin group. The wound healing status on post injury day (POD) 7 and 13 was observed and the wound healing rate was calculated. The wound tissue on POD 7 and 13 was collected for hematoxylin-eosin staining to measure the length of re-epithelialized epidermis and calculate the change rates in diameters of epidermal and dermal wounds, for immunohistochemical staining to detect the relative expressions of keratin 10 and proliferating cell nuclear antigen (PCNA), and for Western blotting to detect the protein expressions of keratin 10 and PCNA. The sample size in all the above experiments was 8 except that in the last experiment was 3. The correlations between the relative expressions of keratin 10 and PCNA in wound tissue in three groups of rats and their wound healing rates, and the correlation between the relative expressions of keratin 10 and PCNA in wound tissue were analyzed.Results:On POD 7, the wound healing rates of rats in diabetes group and diabetes+metformin group were 81.48% (77.89%, 85.53%) and 93.04% (92.51%, 94.24%), which were significantly lower than 100% (97.17%, 100%) in control group (with Z values of 2.37 and -3.36, respectively, P<0.05); the wound healing rate of rats in diabetes+metformin group was significantly higher than that in diabetes group ( Z=3.45, P<0.05). On POD 13, the wound healing rates of rats in control group and diabetes+metformin group were both 100% (100%, 100%), which were significantly higher than 94.47% (90.68%, 99.82%) in diabetes group (with Z values of 2.90 and -2.90, respectively, P<0.05). On POD 7, the change rates in epidermal wound diameter of rats in control group and diabetes+metformin group were significantly higher than that in diabetes group (with Z values of 3.36 and -2.74, respectively, P<0.05). The change rates in dermal wound diameter of rats in the three groups were similar on POD 7 and 13 ( P>0.05). The lengths of re-epithelialized epidermis of rats in control group and diabetes+metformin group on POD 13 were significantly longer than that in diabetes group (with Z values of 3.34 and -2.64, respectively, P<0.05). The relative expressions of keratin 10 in wound tissue of rats in diabetes group on POD 7 and 13 were significantly higher than those in control group (with Z values of -3.36 and -3.26, respectively, P<0.05) and diabetes+metformin group (with Z values of 3.36 and 3.15, respectively, P<0.05), and the relative expression of keratin 10 in wound tissue of rats in diabetes+metformin group on POD 7 was significantly lower than that in control group ( Z=3.05, P<0.05); the relative expressions of PCNA in wound tissue of rats in diabetes group on POD 7 and 13 were significantly lower than those in control group (with both Z values of 3.36, P<0.05) and diabetes+metformin group (with both Z values of -3.36, P<0.05). The protein expressions of keratin 10 in wound tissue of rats in control group and diabetes+metformin group on POD 7 as well as that in diabetes+metformin group on POD 13 were significantly lower than those in diabetes group ( P<0.05), and the protein expressions of PCNA in wound tissue of rats in control group and diabetes+metformin group on POD 7 were significantly higher than that in diabetes group ( P<0.05). There was a significant positive correlation between the relative expression of keratin 10 in wound tissue and the wound healing rate in control group and diabetes+metformin group of rats (with r values of 0.78 and 0.71, respectively, P<0.05), there was a significant negative correlation between the relative expression of PCNA in wound tissue and the wound healing rate in diabetes+metformin group of rats ( r=-0.60, P<0.05), and there was a significant negative correlation between the relative expressions of PCNA and keratin 10 in wound tissue of rats in diabetes group and diabetes+metformin group (with r values of -0.41 and -0.49, respectively, P<0.05). Conclusions:The diabetic rats with full-thickness skin defect wound exhibit delayed healing, accompanied by up-regulation of keratin 10 and down-regulation of PCNA in keratinocytes in the wound tissue. Metformin can promote wound healing in diabetic rats with full-thickness skin defects by down-regulating keratin 10 expression and up-regulating PCNA expression in keratinocytes in the wound tissue, and the wound healing rate was positively correlated with the expression of keratin 10 and negatively correlated with the expression of PCNA.

Objective To compare the lipid-lowering efficacy and safety of fixed-dose combination and free combination of rosuvastatin and ezetimibe in hypercholesterolemia patients who fail to achieve low-density lipoprotein cholesterol(LDL-C)goal with statin monotherapy.Methods A total of 45 hypercholesterolemia patients who switched from statin monotherapy to fixed-dose combination of rosuvastatin and ezetimibe after failing to achieve target LDL-C goal admitted at cardiological departments of First Affiliated Hospital of Sun Yat-sen University,Nanhai Fourth People's Hospital,Foshan First People's Hospital,and Guangdong Provincial People's Hospital between March and June 2024 were enrolled and served as the study group.Another 120 hyper-cholesterolemia patients who treated with free combination of rosuvastatin and ezetimibe were se-lected from Xiamen Regional Health Medical Big Data Platform with propensity score matching and served as control group.The LDL-C level,LDL-C reduction,and changes in TC,HDL-C and TG levels in 4-6 weeks after the medication switch,as well as the safety indicators(AST,ALT,CK,Cre and eGFR)were compared between the two groups.Results In 4-6 weeks after the medication switch,the patients in the study group exhibited a significant decrease in LDL-C level(1.70±0.44 mmol/L vs 2.12±0.87 mmol/L,P＜0.01),obvious LDL-C reduction[(43.17±16.11)％vs(29.14±29.13)％,P＜0.01]when compared to those of the control group.The LDL-C goal attainment rate was significantly higher in the study group than the control group(71.11％vs 45.00％,P=0.003).In addition,there were no statistical differences in the levels of HDL-C and TG and the reductions of HDL-C and TG between the two groups in 4-6 weeks after treatment(P＞0.05).The study group obtained notably lower TC level and TC reduction than the control group in the time(P＜/0.05,P＜0.01).After treatment,no statistical differences were observed between the two groups in terms of AST,ALT,CK,Cre and eGFR(P＞0.05).Conclusion Com-pared to free combination of rosuvastatin and ezetimibe,fixed-dose combination can further reduce LDL-C level in hypercholesterolemia patients who have not achieved LDL-C goal with statin monotherapy,with higher LDL-C goal attainment rate and good safety.

Objective: To analyze the detection rate of sleep problems such as sleep delay and deficiency in preschool children in the middle and lower reaches of the Yangtze River in China,and to provide the reference for the standard of sleeping mode among preschool students. Methods: From October to November 2017, a questionnaire survey was conducted among 27 200 preschool children in 11 cities in Hubei, Anhui and Jiangsu provinces in the middle and lower reaches of the Yangtze River in China. Epidemiology of sleep delays, deficiencies and sleep patterns in preschool children was described. Results: The detection rate of sleep problems in preschool children in the middle and lower reaches of the Yangtze River was 15.3%. Taking the length of sleep and bedtime as the main analysis points, it was found that the average sleeping time point of each age group was 21:31, and the detection rate of bedtime delay was 86.5%. The average length of sleep was (10.60±1.12) hours. The detection rate of sleep deprivation in preschool children was 15.7%. Sleep delay was positively correlated with girls, age increase and parents’ higher educational level (P<0.05), and negatively correlated with living in the city, non-only child and bedroom without TV (P<0.01) .The detection rate of sleep deprivation was positively correlated with children of high age group (4yearold group：OR=1.32，95%CI=1.19-1.46；5-year-old group：OR=2.10，95%CI=1.91-2.32；6-year-old group：OR=2.47，95%CI=2.20-2.77)(P<0.01), and negatively correlated with no TV in bedroom (OR=0.91，95%CI=0.84-0.98) and no light in sleep (OR=0.87，95%CI=0.78-0.97)(P<0.05). Conclusion Preschool children sleep delay and sleep deprivation and other sleep problems are more prominent, affected by family environment and other factors.