Abstract: Objective　To explore the carrying status of four common deafness genes and mutations on 10 loci in newborns in Hainan, and to analyze the molecular epidemiological characteristics of deafness genes and their loci, so as to provide scientific basis for formulating neonatal deafness gene screening strategy and promoting children's hearing health in Hainan. Methods　Newborns born in Hainan from January 2020 to December 2021 were selected as the research objects. The demographic characteristics of the research objects were collected. At the same time, the plantar blood of newborns was collected, and multiplex PCR amplification and directed hybridization combined with high-throughput sequencing technology were applied to detect 10 mutation loci on 4 common deafness genes. T-test or chi square test was used to process the data. Results　A total of 7 124 newborns were included in the study through informed consent, 219 cases of deafness gene mutation were detected with the detection rate of deafness gene of 3.07%. The detection rates of GJB2, SLC26A4, MT-RNR1 and GJB3 were 1.56% (111/7 124), 1.18% (84/7 124), 0.21% (15/7 124) and 0.11% (8/7 124) respectively. Among the 10 loci of the four genes, the positive detection rate of c.235delC locus of GJB2 was the highest, which was 1.38% (98/7 124), followed by c.919-2A>G of SLC26A4 (0.87%, 62/7 124); 2.63% (113/4 289) of the newborns who passed the preliminary hearing screening still carried the deafness gene; in terms of gene type, the detection rate of GJB2 gene in newborns who failed the hearing screening was higher than that in newborns who passed the hearing screening [2.23% (63/7 124) vs 1.12% (48/7 124)，P<0.01]; in terms of gene loci, the detection rate of c.235delC locus in newborns who failed hearing screening was higher than that in newborns who passed hearing screening [2.09% (59/7 124) vs 0.91% (39/7 124)，P<0.01]. Conclusion　The most common deafness genes types in Hainan were GJB2 and SLC26A4; The most common gene mutation sites were c.235delC and c.919-2A>G; 2.63% of the newborns who passed the preliminary hearing screening still carried the deafness gene, among which the high-risk newborns with MT-RNR1 and GJB3 genes were found. Therefore, hearing screening should be combined with deafness gene screening to improve the detection rate of children at high risk of hearing loss.

OBJECTIVEX-linked adrenoleukodystrophy (ALD) is a genetically determined disorder that involves the nervous system white matter, axons, adrenal cortex and testes. The typical clinical manifestations are progressive psychomotor regression, vision and/or auditory impairment and adrenal insufficiency. The clinical manifestation, biochemical change and genetic counseling work of X-linked ALD were analyzed.

METHODSThe clinical features of 29 cases with ALD were summarized and analyzed, including symptoms and signs, measurement of blood very long chain fatty acids (VLCFA), adrenal function, cranial magnetic resonance imaging (MRI) and pedigree investigation.

RESULTSAmong these 29 cases, the clinical phenotype could be classified into childhood cerebral (22 cases), adolescent cerebral (4 cases), adrenomyeloneuropathic (1 case), Addison's disease (1 case) and asymptomatic or presymptomatic (1 case) types. Nine of them had positive family history. Pedigree investigation was consistent with typical sex-linked recessive inheritance. There were 45 ALD patients in these 29 pedigrees. The neurological manifestations varied among members of the same family. Nine cases died during follow up. The causes of death were central respiratory failure or other complications of ALD and so on. Laboratory tests demonstrated abnormally high plasma levels of VLCFA in ALD patients; MRI demonstrated symmetric butterfly-like low T(1) and high T(2) signals in the parieto-occipital white matter. The impairment in the splenium of corpus callosum made the bilateral lesion region converge into one. It could progress anteriorly and injure the bilateral posterior limb of internal capsule and the temporal lobe, and could injure the brainstem inferiorly. Following intravenous injection of contrast material, thin stripe of lacelike enhancement could be observed.

CONCLUSIONSThe atypical initial symptom of ALD was seizures. The MRI showed abnormal signal in the cerebellar white matter. This disease can influence the normal development of children, this was more pronounced in the childhood cerebral ALD type. It tended to progress rapidly with dementia, vegetative state or death. Since antenatal diagnostic method is available now, emphasis should be made on the antenatal examination in order to make an early diagnosis and abort pregnancy if necessary.

Adolescent ; Adrenoleukodystrophy ; blood ; diagnosis ; therapy ; Child ; Child, Preschool ; China ; Fatty Acids ; blood ; Female ; Follow-Up Studies ; Humans ; Male ; Pedigree ; Treatment Outcome

Biopsy ; Biopsy, Needle ; Bone Marrow ; pathology ; Bone Marrow Examination ; methods ; Bone Marrow Neoplasms ; pathology ; secondary ; Breast Neoplasms ; pathology ; Cytological Techniques ; Female ; Humans ; Lung Neoplasms ; pathology ; Male ; Prostatic Neoplasms ; pathology ; Retrospective Studies ; Stomach Neoplasms ; pathology

Objective To study the relationship between Couagen Ⅰ,MMP-2,TIMP-2 gene expression and atrial fibrosis during heart failure(HF)in dog.Methods Fourteen dogs were used and randomized into HF induced by ventricular tachypacing and control group.Burst atrial pacing was used to induce atrial fibrillation(AF).And the mRNA and protein level of collagen Ⅰ,MMP-2 and TIMP-2 were detected by RT-PCR and immunohistochemical technique.Tissue samples were stained with Mallory trichrome.Results Left ventricular ejection fraction (LVEF) decreased from (67.4? 6.0)% to (29.2?7.8)%,the inducible rate of AF(7/7 vs 2/7) and sustained AF(5/7 vs 0/7) increased and duration of AF stabeatrial fibrillation(SAF) [(462.12?181.43)s vs(0.57?0.57) s] prolonged significantly in HF group.Atrial fibrous tissue content and atrial size of HF group were significantly greater than the controls dogs(268.8% in lefe atria and 190.3% in right atria).The mRNA and protein level of collagen Ⅰ(56.2% and 132.2% in lefe atria,37.4% and 78.0% in right atria)and MMP-2 (100.0% and 115.7% in lefe atria,65.7% and 96.8% in right atria) increased evidently in both lefe atria and right atria,TIMP-2 mRNA decreased 46.3% in lefe atria and had no change in right atria and that its protein had no change in both atrium,whereas the ratio of MMP-2/ TIMP-2 of mRNA and protein increased markedly in both lefe atria (285.3% and 148.8%)and right atria (106.1% and 134.7%)of HF group.SAF had a positive correlation with fibrosis and the gene level of collagen Ⅰ in lefe atria,the ratio of MMP-2/TIMP-2 had a positive correlation with fibrosis and collagen Ⅰ gene level in lefe atria during HF.Conclusions The changes of collagen Ⅰ,MMP-2 and TIMP-2 gene expression appear to be a molecular mechanism of AF, and the molecular remodeling of collagen Ⅰ induced by regulation unbalance of MMP-2/TIMP-2 appears to be an important mechanism of atrial fibrosis during HF.

OBJECTIVETo investigate the effect of auxins 2,4-D,IAA,IBA,NAA on induction of adventitious roots as well as that of IBA concentrations on the growth of adventitious roots and the accumulation of caffeic acid derivatives, with test-tube seedling leaves Echinacea pallida as the explant, and cultivate adventitious roots in bioreactors.

RESULT1.0 mg x L(-1) IBA was found the best for the induction of adventitious roots, with the numer of induced adventitious roots up to 22. 5 in each culture dish. Among different concentrations for suspension cultivation of IBA tested, 1.0 mg x L(-1) IBA was found the most suitable for the growth of adventitious roots and the accumulation of caffeic acid derivatives. In a 5 L balloon type bubble bioreactor, 8.98 g x L(-1) dry weight was achieved after one month, which was 2.05 times of 4.38 g x L(-1) dry weight cultivated in a triangular flask. The content of echinacoside cultivated in a bioreactor was 14.08 mg x g(-1) DW, which was 2.4 times of cultivated roots. The contents of chlorogenic acid, chicoric acid and total caffeic acid derivatives were 4.0-25.6 times of ultivated roots.

CONCLUSIONThe study can provide high-quality biomedical drugs containing such caffeic acid derivatives as echinacoside for mass production of Echinacea purpurea medicines.

2,4-Dichlorophenoxyacetic Acid ; pharmacology ; Bioreactors ; Caffeic Acids ; chemistry ; metabolism ; Dose-Response Relationship, Drug ; Echinacea ; drug effects ; growth & development ; metabolism ; Indoleacetic Acids ; pharmacology ; Indoles ; pharmacology ; Naphthaleneacetic Acids ; pharmacology ; Plant Growth Regulators ; pharmacology ; Plant Leaves ; drug effects ; growth & development ; metabolism ; Plant Roots ; drug effects ; growth & development ; metabolism ; Seedlings ; drug effects ; growth & development ; metabolism ; Tissue Culture Techniques ; instrumentation ; methods

Objective To express the plasminogen activator(Pla) of Yersinia pestis and one of its gene fragments,and to detect their immunological reactivity.Methods The pla gene and its specific gene fragment pla-c were amplified by PCR using the EV76 strain as a template.PCR products were then ligated with the plasmid pET32a (+).The recombinant plasmids pET32a (+)-pla and pET32a (+)-pla-c were subsequently trausformed into E.coli BL21 (DE3).The expressed products were purified by HIS affinity chromatography,and their immunological reactivity was detected by Western blotting.Results The recombinant Pla(52.8 × 103) was expressed as inclusion bodies,and the recombinant Pla-c protein (24.0 × 103) was expressed in the soluble form.These two recombinant proteins reacted with anti-Yersinia pestis EV76 rabbit sera.Conclusions The recombinant Pla and its specific fragments have displayed immunological reactivity,and can be served as an alternative diagnosis method for Yersinia pestis.

Objective To study the expression and phosphorylation of focal adhesion kinase(FAK) in rat's au- tologous vein graft and the olmesartan modulating effect.Methods Autologous external jugular veins were grafted to common carotid arteries in 40 male Sprague Dawley rats.After surgery,rats were randomly assigned to the fol- lowing groups:sham;control;olmesartan treatment(10mg/kg.d by gavage);or physiological saline.The intimal thickness,the I/M in vein grafts was quantitated by HE stain.The expression and phosphorylation of focal adhe- sion kinase were assessed by Western-blotting,PCNA and ?-smooth muscle actin were measured by immunohisto- chemistry.Results Neointimal hyperplasia in control group was characterized by significantly increased intimal thickeness I/M(P

OBJECTIVETo identify the effects of NMDA receptor subunits NR2A and NR2B expression in rat's hippocampus after exposure to 1800 MHz radiofrequency radiation.

METHODSFour-week old female Wistar rats were randomly divided into four groups, with 12 animals for each. The subjects in two experimental groups had been continuously exposed to 1800 MHz microwave radiation (CW) with respective power density of 0.5 mW/cm(2) and 1.0 mW/cm(2) 12 hours each day for 21 days. Meanwhile, sham-controls were carried out. The brain tissue sections were performed by immunohistochemistry to demonstrate both expressions of NR2A, NR2B immune-activity in the hippocampal CA1, CA3 and DG by using computer-assisted image analysis system.

RESULTSIn NR2A: the expression of 0.5 mW/cm(2) power density group was significantly lower than 0 mW/cm(2) power density group in CA3 [(8.5 +/- 1.5) vs (11.1 +/- 1.8), P < 0.01] and had not been significantly changed in CA1 and DG. The expression of 1.0 mW/cm(2) power density group was significantly lower than 0 mW/cm(2) power density group in CA1 and CA3 [(7.9 +/- 1.6) vs (9.7 +/- 1.5); (8.4 +/- 1.7) vs (11.1 +/- 1.8), respective P < 0.05, P < 0.01] and had not been significantly changed in DG. In NR2B: the expression of 0.5 mW/cm(2) power density group was significantly lower than 0 mW/cm(2) power density group in CA1 and CA3 [(16.4 +/- 1.0) vs (17.8 +/- 1.6); (9.6 +/- 1.9) vs (11.2 +/- 2.1), respective P < 0.05]. The expression of 1.0 mW/cm(2) power density group was significantly lower than 0 mW/cm(2) power density group in CA1, CA3 and DG [(13.1 +/- 2.4) vs (17.8 +/- 1.6); (9.3 +/- 1.4) vs (11.2 +/- 2.1); (7.3 +/- 0.1) vs (8.5 +/- 1.0), respective P < 0.01, P < 0.05, P < 0.05].

CONCLUSIONThere were findings of the effects on NMDA receptor subunits in different hippocampus sections after exposure to 1800 MHz radiofrequency radiation.

Animals ; Female ; Hippocampus ; metabolism ; radiation effects ; Immunohistochemistry ; Radio Waves ; Rats ; Rats, Wistar ; Receptors, N-Methyl-D-Aspartate ; biosynthesis

Objective To conduct a microbiological and parasitological investigation of experimental minipigs in Guangdong province. Methods Four major experimental minipig production units in Guangdong province were included in this investigation. Samples were taken from a total of 154 pigs of 4 brreds, i. e. , Bama minipigs, Juema minipigs, Tibet minipigs and Wuzhishan minipigs. Pig fur, scales, serum, rectal swabs and feces samples were collected for detection of 20 pathogens. The data were analyzed and compared among the production units and breeds. Results Mixed infections were detected in all the four institutions. The infection rates of 7 pathogens were rather high: Streptococcus suis type 2 (50. 7%), Actinobacillus pleuropneumoniae (40. 3%), Mycoplasma hyopneumoniae (100%), Japanese encephalitis virus (41. 3%), porcine circovirus type 2 (74. 8%), porcine transmissible gastroenteritis virus (73. 8%),gastroenteritis virus (44. 7%). Porcine parvovirus (26. 0%), pseudorabies virus(15. 6%) and intestinal worms (3. 2%) were also detected in some animals. The immune qualified rates of classical swine fever virus (62. 8%) and foot-and-mouth disease virus (35. 8%) were rather low. The immune qualified rate of pseudorabies virus was as high as 98. 4%. Besides, Salmonella, Brucella, swine dysentery snake like spirochetes, dermatophytes, influenza virus. Toxoplasma gondii, ectoparasites, and coccidia were not detected. Conclusions The results of this investigation indicate that epidemiological quality control of pathogens in experimental minipigs and efforts to establish high grade minipig population in Guangdong province remain to be strengthened. Our study also provides a basis for revision of local and even national standards for experimental minipigs.

OBJECTIVETo detect p15(INK4B) methylation levels and the kinetics of the methylation status before and after decitabine to explore its influences on prognosis and response to decitabine in myelodysplastic syndromes (MDS) patients.

METHODSWe examined 261 MDS patients (143 male and 118 female) with the median age of 52 years (32-78). Of them, 172 cases were low-risk group (low-risk 104 cases, intermediate-1 68 cases), 89 cases high-risk group (intermediate-2 52 cases, high risk 37 cases). Collections of bone marrow mononuclear cells of MDS patients and extracted the genomic DNA, the methylation status of p15(INK4B) was detected by methylation-specific PCR (MSP) method. Survival analysis was conducted according to the level of p15(INK4B) methylation in the cohort of patients. The kinetics of the methylation levels of p15(INK4B) in 58 MDS patients before and after 2 courses of decitabine have been assessed with the method of MSP.

RESULTSThe methylation level of p15(INK4B) in low-risk group patients was significantly lower than that in high-risk group (117.22 vs 157.63, P<0.05 ). The expected 2-year survival rate of p15(INK4B) methylation positive patients was lower than that of negative ones (91.8% vs 69.8%, P<0.05); the expected 2-year survival rate of p15(INK4B) methylation positive patients was shorter than that of negative ones in low-risk group（78.2% vs 92.0%, P<0.05）, meanwhile there was no significant difference in terms of expected 2-year survival rate and median expected survival between p15(INK4B) methylation positive and negative patients in high-risk group [35.6% vs 38.5%, (17.0±9.3) month vs (18.0±5.7) month, P>0.05]. Multivariate analysis showed p15(INK4B) methylation degree was an independent prognostic factor for overall survival. No statistical difference of overall response (OR) rates were found between p15(INK4B) methylation positive patients and negative patients before decitabine（65.9% vs 76.5%, P>0.05）, and complete remission (CR) rates between these two groups also showed no statistical difference（22.0% vs 29.4%, P>0.05）. p15(INK4B) methylation levels had no obvious change before and after treatment in decitabine responders（P>0.05）.

CONCLUSIONThe survival of newly diagnosed MDS patients with positive p15(INK4B) methylation was comparatively shorter, but p15(INK4B) methylation had no influence on response to decitabine.

Adult ; Aged ; Azacitidine ; analogs & derivatives ; Cyclin-Dependent Kinase Inhibitor p15 ; genetics ; DNA Methylation ; Female ; Humans ; Male ; Middle Aged ; Myelodysplastic Syndromes ; drug therapy ; genetics ; Prognosis ; Survival Rate ; Treatment Outcome