Objective To investigate the effects of bone mesenchymal stem cell (BMSC) transplantation on neuronal nuclei (NeuN) and neurogenin 1 (Ngnl) in focal cerebral ischemic rats.Methods A total of 64 healthy adult male Sprague-Dawley rats were randomly divided into normal (N) + phosphate-buffered solution (PBS),middle cerebral occlusion (MCAO)+ PBS,N + BMSC and MCAO + BMSC groups (n =16 in each group).A rat model was induced by the intraluminal suture method.BMSC was cultured in vitro.At 24 h after modeling,brain transplantation was conducted.Magnetic resonance imaging (MRI) was used to detect infarct volume in vivo.NeuN/DAP,Ngnl/DAPIimmunofluorescence double-labeling and Western blot were used to detect the expression of NeuN and Ngnl around ischemic brain tissue.Results On day 14 after transplantation,the T1-and T2-weighted imaging revealed that the cerebral cortex and striatum had abnormal signal areas in the rats of the MCAO group.The infarct volume of the MCAO + BMSC group was significantly less than that of the MCAO + PBS group (32.5％ ± 4.2％ vs.47.9％ ± 7.9％ ; P ＜ 0.01).Immunofluorescence doublelabeling assay showed that the numbers of cells of NeuN+/DAPI+ (976.2 ± 87.5/mm2 vs.1 908.3 ±127.8/mm2; P ＜ 0.01) and Ngn1 +/DAPI + (251.6 ± 23.1/rmm2 vs.285.1 ± 25.2/mm2 ; P ＜ 0.01) of the MCAO + PBS group were significantly less than those of the N + PBS group,but those of NeuN+/DAPI +(1 439.9 ± 101.7/mm2; P ＜ 0.01) and Ngn1 +/DAPI + (356.3 ± 35.6/mm2; P ＜ 0.01) of the MCAO + BMSC group were significantly more than the MCAO + PBS group.Western blot analysis showed that the protein expression levels of NeuN (0.69 ±0.06 vs.0.91 ±0.09; P ＜0.01) and Ngn1 (0.53 ±0.05 vs.0.62 ±0.07;P ＜0.01) of the MCAO +PBS group were significantly lower than those of the N +PBS group,but those of NeuN (0.82 ± 0.07; P ＜ 0.01) and Ngn1 (0.77 ± 0.09; P ＜ 0.01) of the MCAO + BMSC group were significantly higher than the MCAO + PBS group.Conclusions BMSC transplantation may promote the expression of NeuN and Ngn1,and alleviate MCAO caused brain injury.

Objective: To analyze the influencing factors for bladder cancer in Taizhou,and to provide evidence for strengthening the prevention and treatment of bladder cancer in Taizhou. Methods : A total of 500 cases of bladder cancer diagnosed in Taizhou Hospital from 2012 to 2017 were selected as a case group,and 504 patients without tumor or urinary system diseases during the same period were selected as a control group. A structured questionnaire was used to retrospectively investigate the demographic information,occupational exposure（whether they were exposed to aromatic amine,polycyclic aromatic hydrocarbons,tobacco,tobacco smoke or heavy metals at work）,healthy behaviors and diets of the two groups one year before admission. A Logistic regression model was used to analyze the influencing factors for bladder cancer. Results : There was no significant differences in sex,age,ethnicity,education and marital status between the case group and the control group（P > 0.05）. Patients with occupational exposure history accounting for 31.60% in the case group and 24.60% in the control group,for overweight/obesity were 37.60% and 31.74%,for smoking were 55.80% and 46.23%,for high vegetable intake frequency were 43.80% and 52.58%,for high fruit intake frequency were 55.40% and 62.70%,for physical activity were 24.60% and 31.75%,respectively. The results of multivariate logistic regression analysis showed that occupational exposure（OR=1.861,95%CI：1.229-2.836）,overweight/obesity（OR=1.374,95%CI：1.021-1.863）,current smoking（OR=1.664,95%CI：1.101-2.503）and previous smoking（OR=1.454,95%CI：1.016-2.066）were the risk factors for bladder cancer. High vegetable intake frequency（OR=0.731,95%CI：0.566-0.947）,high fruit intake frequency（OR=0.659,95%CI：0.463-0.927）and vigorous physical activity（OR=0.566,95%CI：0.403-0.798）were the protective factors for bladder cancer. Conclusion Occupational exposure,overweight/obesity,current smoking,previous smoking were the risk factors for bladder cancer. High vegetables intake frequency,high fruit intake frequency and vigorous physical activity were the protective factors for bladder cancer.

Objective To investigate the effect of small interfering RNA (siRNA) interference in the expression of epidermal growth factor receptor (EGFR) on the radiosensitivity of esophageal squamous carcinoma (Eca-109) and esophageal adenocarcinoma (OE-19) cell lines.Methods Human Eca-109 and OE-19 cell lines were selected as study subjects.Various EGFR-siRNA and negative siRNA were synthesized chemically through lipofection.Reverse transcription-polymerase chain reaction and Western blot were applied to measure the expression of EGFR before and after transfection,and the CCK8 assay was applied to analyze the influence of transfection on cell proliferation.Blank control groups of Eca-109 and OE-19 cells (O1 and O2 groups),simple irradiation groups (R1 and R2 groups),and EGFR-siRNA irradiation groups (E-R1 and E-R2 groups) were set,and the doses for single irradiation were 0,2,4,6,and 8 Gy.The colony-forming assay was applied to calculate survival fraction (SF) and sensitization enhancement ratio (SERD0 ratio),and flow cytometry was applied to evaluate the influence of EGFR-siRNA combined with radiotherapy on cell cycle distribution and apoptosis rate,and the dose for single irradiation was 6 Gy.Results The expression of EGFR in both cell lines was significantly down-regulated by EGFR-siRNA,and the inhibition rate of cell proliferation by transfection was＜5％ (4.9％ and 4.5％,respectively).The results of colony-forming assay showed that the cells in the E-R1 and E-R2 groups had a lower SF than those in the O1 and O2 groups,with an SERD0ratio of 1.40 and 1.01,respectively.The results from flow cytometry showed that compared with the E-R2 group,the E-R1 group had a higher proportion of cells in G2/M phase and a lower proportion of cells in S phase after irradiation (P=0.016 and 0.028),as well as a higher apoptosis rate (P=0.007).Conclusions Compared with the cell line OE-19,the cell line Eca-109 has a significantly increased radiosensitivity when treated with siRNA interference in EGFR expression.

Objective To investigate the effect of insulin and gliclazide therapy on endoplasmic reticulum (ER) stress and insulin sensitivity in the liver of type 2 diabetic rats.Methods A high fat diet plus a low-dose of streptozotocin was implemented to create a type 2 diabetic rats which were randomly divided into diabetes mellitus (DM) group,insulin treatment (INS) group and gliclazide treatment (GT)group; and healthy rats were as normal control group.Diabetic rats in INS and GT groups were given neutral protamine hagedorn (NPH) insulin and gliclazide respectively for 3 weeks.Protein expression levels of immunoglobulin binding protein (Bip),spliced X-box binding protein 1 (XBP-ls),phosphorylated c-Jun on serine 73 (p-c-Jun),phosphorylated insulin receptor substrate 1 on serine 307 (p-IRS-1),and glucose-6-phosphatase (G6Pase) in liver homogenate were detected by Western blotting.Results Compared with the normal rats,Bip and XBP-Is in the DM group were up-regulated (0.28 ±0.07 vs 0.90 ±0.10 for Bip;0.41 ± 0.07 vs 0.95 ±0.07 for XBP-1 s; both P ＜ 0.01 ) ; p-c-Jun (0.59 ± 0.18 vs 1.94 ± 0.03 ),p-IRS-1( 1.73 ± 0.18 vs 5.32 ± 0.22) and G6Pase (0.11 ± 0.01 vs 0.45 ± 0.01 ) were increased ( all P values ＜0.01 ).In the INS group,all of aforementioned changes were reversed (0.90 ± 0.10 vs 0.25 ± 0.04 for Bip; 0.95 ±0.07 vs 0.47 ±0.01 for XBP-1s; 1.94 ± 0.03 vs 0.50 ±0.10 for p-c-Jun; 5.32 ± 0.22 vs 1.59 ±0.32 for p-IRS-1 ; 0.45 ±0.01 vs 0.15 ±0.02 for G6Pase,all P values ＜0.01 ).In the GT group,all of aforementioned changes were also attenuated ( 0.90 ± 0.10 vs 0.53 ± 00.02 for Bip ; 0.95 ± 0.07 vs 0.78±0.02 for XBP-1s; 1.94 ±0.03 vs 1.33 ±0.11 for p-c-Jun; 5.32 ±0.22 vs 3.13 ±0.02 for p-IRS-1; 0.45 ± 0.01 vs 0.25 ± 0.01 for G6Pase,all P values ＜ 0.05).Furthermore,all of aforementioned protein levels were down-regulated more obviously in the INS group comparing to the GT group ( all P values ＜ 0.01 ).Conclusions Both insulin and gliclazide therapy could relieve ER stress and e-Jun N-terminal kinase activity and improved insulin sensitivity.The effect of insulin on Bip,XBP-1s,p-c-Jun,p-IRS-1 and G6Pase protein expressions is more obvious than that of glilcazide,which indicates besides lowering glucose,insulin might have protective effects of anti-inflammation,anti-oxidative stress or stimulation of lipid redistribution.

Objective: To identify potential variant in a child diagnosed as infantile neuroaxonal dystrophy. Methods: Genomic DNA was extracted from peripheral blood samples from the patient and his parents and subjected to next generation sequencing. Suspected variant was verified by PCR and Sanger sequencing. Pathogenicity of the mutation was predicted by using bioinformatic software including SIFT and PolyPhen-2. Results: The child was found to carry compound heterozygous variations c. 668C>A (p.Pro223Gln) and c. 2266C>T (p.Gln756Ter) of the PLA2G6 gene, which were respectively inherited from his father and mother. c. 2266C>T has changed codon 756 (glutamine) into a stop codon, resulting premature termination of peptide chain synthesis. c. 2266C>T has not been reported previously and was predicted to be harmful. Conclusion The compound variants of c. 668C>A (p.Pro223Gln) and c. 2266C>T (p.Gln756Ter) of the PLA2G6 gene probably underlies the disease in the child. Above finding has enriched the variant spectrum of the PLA2G6 gene.

OBJECTIVETo investigate the gene frequencies and mutation patterns of alpha thalassemia (alpha-thal) and beta thalassemia (beta-thal) in Liuzhou city of Guangxi Zhuang Autonomous Region.

METHODSCluster sampling was used. A total of 1 028 of umbilical blood samples were collected for a prevalence study of alpha-thal and a total of 1 312 healthy young people when receiving pre-marriage consultation were recruited for a beta-thal prevalence survey. Individuals live in city or town area of Liuzhou. A complete blood count as well as hemoglobin electrophoresis analysis were done in all of samples for phenotyping of alpha and beta-thals. Those with Hb Bart's for alpha-thal indicator and those with both microcytosis (MCV < 85 fl) and elevated levels of Hb A(2) (>/=4.0%) for beta-thal were further studied by DNA analysis. PCR-based methodologies were used to characterize the mutation contributions of alpha and beta-thals. All the subjects were tested for the state of carrying beta-thala alleles for evaluating the situation of the compound heterozygotes of alpha-thal with beta-thal.

RESULTSOf 1 028 random samples of umbilical blood screened, 112 of subjects were defined to be the gene carriers of alpha-thal. The alpha-thal carrier rate was as high as 11.19% including 3 compound heterozygotes. Five well-known types of alpha-thal alleles were detected with gene contributions of 37.4% (--(SEA) deletion), 31.3% (-alpha(3.7) deletion), 17.4% (-alpha(4.2) deletion), 12.1% (alpha(CS)alpha mutation), and 0.9% (alpha(QS)alpha mutation), successively. Of the 1 312 adult specimens studied, 89 with beta-thal including 14 of the compound higher Hb F subjects were detected. All of the 89 phenotypic beta-thal carriers had the mutations in the beta-globin gene, making the overall prevalence 6.78%. The commonly seen three mutations, beta CD41 - 42 (-CTTT) frameshift, beta CD17 (T-A) nonsense mutation and beta-28 (A-G) promoter variation were accounted for 90% of the beta-thal alleles in Liuzhou. Of these beta-thal subjects, 16 (accounting for 18%) were found to be the compound heterozygosity for a beta-thal and an alpha-thal with 9 different types of gene defects with a detection rate 1.22%.

CONCLUSIONData from ecidation of alpha and beta-thal gene frequencies and mutation spectrum in Liuzhou city was useful for genetic counselling and prenatal diagnosis of this disease.

Adult ; China ; epidemiology ; Female ; Gene Frequency ; Genetic Counseling ; Humans ; Male ; Prevalence ; alpha-Thalassemia ; epidemiology ; genetics ; beta-Thalassemia ; epidemiology ; genetics

Objective: To analyze variations of TYR and P genes among 14 patients with clinically diagnosed oculocutaneous albinism. Methods: Potential variations of the TYR and P genes were detected by Sanger sequencing. Novel variations were predicted with bioinformatics software including SIFT and PolyPhen-2. Results: No variation was found in the TYR gene, while 9 types of variations were found in the P gene among the 14 patients, which included c. 803-3C>G (7/26), c. 1327G>A (p.Val443Ile) (5/26), c. 632C>T (p.Pro211Leu) (4/26), c. 1832T>C (p.Leu611Pro) (3/26), c. 1349C>A (p.Thr450Lys) (2/26), c. 2363C>T (p.Ser788Leu) (2/26), c. 2228C>T (p.Pro743Leu) (1/26), c. 1525A>G(p.Thr509Ala) (1/26), and c. 1349C>T(p.Thr450Met) (1/26). Only 1 heterozygous variation was detected in 2 families. c. 2363C>T (p.Ser788Leu), c. 1832T>C (p.Leu611Pro) and c. 1525A>G (p.Thr509Ala) were not reported previously and predicted as "harmful" to the protein function. Conclusion The main type of ocular albinism is oculocutaneous albinism type Ⅱ in Liuzhou region, where the most common variations of the P gene were c. 803-3C>G and c. 1327G>A (p.Val443Ile). Above finding has enriched the variation spectrum of the P gene.

OBJECTIVE: To detect variants of IVD gene among 4 neonates with suspected isovalerate acidemia in order to provide a guidance for clinical treatment. METHODS: 111 986 newborns and 7461 hospitalized children with suspected metabolic disorders were screened for acyl carnitine by tandem mass spectrometry. Those showing a significant increase in serum isovaleryl carnitine (C5) were analyzed for urinary organic acid and variants of the IVD gene. RESULTS: Four cases of isovalerate acidemia were detected, which included 2 asymptomatic newborns (0.018‰, 2/111 986) and 2 children suspected for metabolic genetic diseases (0.268‰, 2/7461). The formers had no obvious clinical symptoms. Analysis of acyl carnitine has suggested a significant increase in C5, and urinary organic acid analysis has shown an increase in isovaleryl glycine and 3-hydroxyisovalerate. Laboratory tests of the two hospitalized children revealed high blood ammonia, hyperglycemia, decreased red blood cells, white blood cells, platelets and metabolic acidosis. The main clinical manifestations have included sweaty foot-like odor, feeding difficulty, confusion, drowsiness, and coma. Eight variants (5 types) were detected, which included c.158G>A (p.Arg53His), c.214G>A (p.Asp72Asn), c.548C>T (p.Ala183Val), c.757A>G (p.Thr253Ala) and 1208A>G (p.Tyr403Cys). Among these, c.548C>T and c.757A>G were unreported previously. None of the variants was detected by next generation sequencing of 2095 healthy newborns, and all variants were predicted to be likely pathogenic based on the guidelines from the American College of Medical Genetics and Genomics. CONCLUSION The incidence of isovalerate acidemia in Liuzhou area is quite high. Screening of metabolic genetic diseases is therefore recommended for newborns with abnormal metabolism. The discovery of novel variants has enriched the mutational spectrum of the IVD gene.
Infant, Newborn ; Child ; Humans ; Acidosis ; Carnitine ; Erythrocytes ; High-Throughput Nucleotide Sequencing

Objective:To study the distribution and antibiotics resistance of the main pathogens of neonatal purulent meningitis in different regions of China.Methods:A retrospective descriptive clinical epidemiological study was conducted in children with neonatal purulent meningitis which admitted to 18 tertiary hospitals in different regions of China between January 2015 to December 2019. The test results of blood and cerebrospinal fluid, and drug sensitivity test results of the main pathogens were collected. The distributions of pathogenic bacteria in children with neonatal purulent meningitis in preterm and term infants, early and late onset infants, in Zhejiang Province and other regions outside Zhejiang Province, and in Wenzhou region and other regions of Zhejiang Province were analyzed. The chi-square test was used for statistical analysis.Results:A total of 210 neonatal purulent meningitis cases were collected. The common pathogens were Escherichia coli ( E. coli)(41.4%(87/210)) and Streptococcus agalactiae ( S. agalactiae)(27.1%(57/210)). The proportion of Gram-negative bacteria in preterm infants (77.6%(45/58)) with neonatal purulent meningitis was higher than that in term infants (47.4%(72/152)), and the difference was statistically significant ( χ2=15.54, P=0.001). There were no significant differences in the constituent ratios of E. coli (36.5%(31/85) vs 44.8%(56/125)) and S. agalactiae (24.7%(21/85) vs 28.8%(36/125)) between early onset and late onset cases (both P>0.05). The most common pathogen was E. coli in different regions, with 46.7%(64/137) in Zhejiang Province and 31.5%(23/73) in other regions outside Zhejiang Province. In Zhejiang Province, S. agalactiae was detected in 49 out of 137 cases (35.8%), which was significantly higher than other regions outside Zhejiang Province (11.0%(8/73)). The proportions of Klebsiella pneumoniae, and coagulase-negative Staphylococcus in other regions outside Zhejiang Province (17.8%(13/73) and 16.4%(12/73)) were both higher than those in Zhejiang Province (2.9%(4/137) and 5.1%(7/137)). The differences were all statistically significant ( χ2=14.82, 12.26 and 7.43, respectively, all P<0.05). The proportion of Gram-positive bacteria in Wenzhou City (60.8%(31/51)) was higher than that in other regions in Zhejiang Province (38.4%(33/86)), and the difference was statistically significant ( χ2=6.46, P=0.011). E. coli was sensitive to meropenem (0/45), and 74.4%(32/43) of them were resistant to ampicillin. E. coli had different degrees of resistance to other common cephalosporins, among which, cefotaxime had the highest resistance rate of 41.8%(23/55), followed by ceftriaxone (32.4%(23/71)). S. agalactiae was sensitive to penicillin, vancomycin and linezolid. Conclusions:The composition ratios of pathogenic bacteria of neonatal purulent meningitis are different in different regions of China. The most common pathogen is E. coli, which is sensitive to meropenem, while it has different degrees of resistance to other common cephalosporins, especially to cefotaxime.

Objective: To explore the association of TBX5 polymorphisms and environmental exposure index with susceptibility to oral cancer. Methods: A case-control study was conducted to collect 300 oral cancer patients hospitalized in the Department of Oral and Maxillofacial Surgery, the First Affiliated Hospital of Fujian Medical University from September 2010 to December 2016. A total of 445 non-tumor patients were selected as the control group. Questionnaires were used to collect the information of all subjects and 5 ml peripheral blood was collected to detect single nucleotide polymorphisms (SNPs) of the rs10492336 locus of TBX5 gene. According to the environmental exposure index score, subjects were divided into two groups, low risk group (0-2.31) and high risk group (2.32-11.76). To analyze the association of TBX5 gene rs10492336 SNPs, environmental exposure index and oral cancer and its interactions. Results: The age of all subjects in the case group and control group were (56.19±13.10) years and (54.56±12.48) years old. Compared with CC genotype, the OR (95%CI) values of the co-dominant genetic model AC genotype and the dominant genetic model AC+AA genotype were 0.69 (0.49-0.98) and 0.70 (0.51-0.97), respectively. Compared with the low risk group, the OR (95%CI) risk of oral cancer in the high risk group was 3.72 (2.55-5.43). The results of gene-environment interaction analysis showed that compared with the group with CC genotype and high risk of environmental exposure index, the OR (95%CI) value of oral cancer in the group with AC+AA genotype and low risk of environmental exposure index was 0.18(0.10-0.31). Furthermore there was a multiplicative interaction between rs10492336 SNPs and environmental exposure index (β=-0.405, P<0.001). Conclusion This study suggests that the TBX5 gene rs10492336 SNPs and environmental exposure index were associated with oral cancer. And there was a multiplication interaction between rs10492336 SNPs and environmental exposure index.