Objective To establish a rapid method to detect mutations in rpoB genes of rifampin-resistant Mycobacterium tubereulosis in dinical specimens using Real-time fluorescence PCR molecular beacon assay.Methods 174 strains of Mvcobacterium tuberculosis clinical isolates were analyzed using real-time fluorescence PCR molecular beacon assay foilowed with DNA sequencing while 12 strains of NTM and 4 strains of bacteria other than Mycobacterium tuberculosis were used as the contrast.Results Eighty-two 89.1 of 92 rifampin (RIF)-resistant strains and 3 of 82 RIF-sensitive strains were found to harbor mutation in the rpoB gene using real-time fluorescence PCR-molecular beacon assay.The specificity, sensitivity,and accuracy of this assay were 96.3%,89.1%,and 92.5%,respectively-Eithty-three of 92 RIF-resistant strains and 1 of 82 RIF-sensitive strains were found to harbor mutation in the rpoB gene using the direct DNA sequencing.The specificity,sensitivity,and accuracy of the direct DNA sequencing were 98.8,90.2%,and 94.2%,respectively.As compared with real-time PCR molecular beacon assay,171 of 174(98.3%)strains of myeobactefium tuberculosis clinical isolates had the salne results.Conclusion Real-time fluorescence PCR-molecular beacon assay can be used as a rapid screen method to detect RIF-resistant isolates.

Purpose To investigate the relationship and clinical significance of high-risk human papillomavirus infection and expression of c-jun and c-fos protein in cervical carcinoma.Methods Cervista HR-HPV testing,immunohistochemical SP method were employed to detect the infection of HR-HPV,c-jun and c-fos protein in 70 cases of CSCC,60 cases of cervical intraepithelial neoplasia and 20 cases of chronic cervicitis.The correlation between c-jun and c-fos protein expression and of HR-HPV infection was analyzed.Results Of the 70 cases of CSCC,the positive HR-HPV was 69 cases,the positive rate in HR-HPV in A9 group was the highest (85％,59/69).The positive expression of c-jun and c-fos were 80％ (56/70),85.7％ (60/70) in 70 cases CSCC,70％ (21/30) and 70％ (21/30) in 30 cases of cervical high grade intraepithelial neoplasia,and 20％ (6/30),23.3％ (7/30) in 30 cases cervical low grade intraepithelial neoplasia,but the positive expression rates of c-jun and c-fos in the 20 cases of chronic cervicitis were 0.Expression of c-jun and c-fos in cervical cancer group was higher than that in the chronic cervicitis group and low grade intraepithelial neoplasia (P ＜ 0.05).The expression of c-jun and c-fos was statistically significant in different groups of clinical stage and pathologic grading in the CSCC (P ＜ 0.05).Additionally,the expression of c-jun and c-fos was positively correlated with the infection of HR-HPV in CSCC (P ＜ 0.01).Conclusion The infection of HR-HPV has significant subtype-specific and has a positive correlation with the expression of c-jun and c-fos in CSCC,which suggests that AP-1 pathway activation after HRHPV infection may be associated with the occurrence and development of cervical carcinoma.

[Summary] In this study, PubMed, Embase, China National Knowledge Infrastructure, databases VIP Chinese Periodical Database, and Wanfang Chinese Periodical Database were systematically searched for the case-control study related β3-adrenergic receptor ( ADRB3 ) Trp64Arg gene polymorphism to overweight/obesity among children from 1962 to 2014.Twelve eligible studies with 2 222 overweight/obese children and 1 955 normal children were included according the uniform inclusion and exclusion criteria.Meta-analyses showed that Trp64Arg polymorphism was associated with significantly increased overweight/obesity risk in Arg carriers among children( OR=1.34,95%CI1.17-1.53).Afterstratificationforethnicity,highlysignificantcoorelationofTrp64Argpolymorphism to overweight/obesity in Asian children(OR=1.44, 95%CI 1.23-1.68) but not significant in Europe(OR=1.05, 95%CI 0.79-1.40).It suggested that Trp64Arg polymorphism is associated with overweight/obesity susceptibility in children.Our results support an strong association between Trp64Arg polymorphism and overweight/obesity among the Asian children investigated.

Objective:To explore the possible mechanism of astragaloside involved in the mouse podocytes injury induced by TGF-β1 in vitro.Methods:Mouse podocytes were cultured in vitro and then all cell were divided into 5 groups:normal control group , TGF-β1 treatment group ,TGF-β1 treatment +astragaloside low dose group ,TGF-β1 treatment +astragaloside middle dose group and TGF-β1 treatment +astragaloside high dose group.The proliferation rate of each group was investigated by MTT assay ,the expression of TRPC6 protein and mRNA were measured by Western blot and RT-PCR respectively after 48 hours.Results:TGF-β1 can significantly inhibit the proliferation of podocytes ( P<0.05) ,fusions of foot processes or even effaced of podocytes were observed .TGF-β1 could also increase the expression of TRPC6.Astragaloside could reduce the inhibition of TGF-β1 to the proliferain of podocytes significantly ,make the cell shape tend to be normal,and reduce the expression of TRPC6 mRNA and protein with dose-effect relation.Conclusion:TRPC6 play an impor-tant role in the TGF-β1 induecd podocytes injury .Astragaloside can alleviate podocytes injury by reduce the expression of TRPC 6.

Objective To investigate the roles of TrkA and TrkB in radiation-induced hippocampal neurogenesis impairment.Methods Fifty-six rats were randomized into radiation group and sham control group.Radiation group received whole brain irradiation at a single dose of 10 Gy.The hippocampus were separated from rats in day 1,day 3,day 14 and 1 month after irradiation.Western blot and RT-PCR were applied to detect the protein levels and mRNA levels.Golgi staining was used to observe the dendritic spine of hippocampus.Immunofluorescence was performed to detect neural precursor's proliferation.Results Compared with control group,the numbers of dendritic spine significantly decreased after irradiation and its shape change obviously.Immunofluorescence showed a significant decrease in neural precursor's proliferation comparing with control group (t =6.49,P ＜ 0.05).Protein level of TrkA expression increased (t =2.64,3.06,4.80,2.64,P ＜ 0.05),while the levels of TrkB protein expression decreased significantly (t =4.59,3.06,2.81,2.57,P ＜ 0.05).The mRNA level of TrkA expressions increased (t =4.57,3.06,5.39,5.86,P ＜ 0.05),while the mRNA level of TrkB decreased (t =14.87,11.69,4.98,P ＜ 0.05).Conclusions As a signaling pathways downstream of NGF and BDNF,TrkA and TrkB may play an important role in radiation-induced neurogenesis impairment.

Objective:To observe the effects of continuous light exposure on skeletal muscle fiber type transformation and lipid metabolism, and to explore its internal relationship.Methods:Mice were randomly divided into normal light group and 24-hour continuous light group by random number table. The serum and skeletal muscle lipid content and urine 6-sulfatoxymelatonin(6-SML)level were detected by ELISA. The expression of circadian clock and lipid metabolism related genes mRNA were observed by realtime PCR. The muscle fiber type and lipid deposition were evaluated by tissue immunofluorescence as well as oil red O staining.Results:Compared with the normal light group, the level of 6-SML in urine at night decreased( P<0.05), and the expression level and rhythm of brain and muscle ARNT-like protein 1(Bmal1), circadian locomotor output cycles protein kaput(Clock), and period 2(Per2)mRNA in the skeletal muscle changed in continuous light group. In addition, the body weight, blood lipid, free fatty acid, and triglyceride contents of skeletal muscle in continuous light group increased significantly( P<0.05 or P<0.01), the expression of carnitine palmitoyltransferase 1b (Cpt1b)mRNA, the key enzyme of fatty acid oxidation, decreased significantly( P<0.05), while the expression of stearoyl-CoA desaturase(Scd1)mRNA, a lipid synthesis related gene, increased significantly( P<0.01). Further immunofluorescence analysis showed that the proportion of slow muscle fibers decreased and that of fast muscle fibers increased in continuous light group(both P<0.05). Conclusion:The process of ectopic deposition of lipid in skeletal muscle in mice induced by continuous light exposure may be related to the remodeling of skeletal muscle fibers.

OBJECTIVETo investigate the pathological course in intracranial aneurysms.

METHODSNormal intracranial artery tissue (cortex fistulization) from 1 case, ruptured aneurysms tissuses from 11 cases, unruptured aneurysm tissues from 2 cases were obtained by neurosurgical excision. Routine HE staining was used to observe histological characteristics. In situ hybridization was used to observe the expression of the monocyte chemoattractant protein-1 (MCP-1) mRNA in the walls of the normal artery and aneurysms.

RESULTSBy the HE staining showed that the wall of the ruptured aneurysms (10 cases) and unruptured ones (2 cases) had increased intima and connectivum extima. The fibroblast in the intima was arrayed in the disorder. Monocyte-like cells can be seen in the whole aneurysm wall. In one case aneurysms wall (ruptured) glass-like fiber structure was left over, few cells could be seen. In 9 cases, mural thrombus was found. The thrombus represented with organization. In situ hybridization, MCP-1 mRNA was not detectable in the normal artery. The hybridization signal could be observed in the ruptured aneurysms (10 cases) and unruptured ones (2 cases) often in the intima. MCP-1 mRNA appeared to be expressed by fibroblast cells in its cytoplasm. Monocyte-like cells had little cytoplasm, and the signal was seldom seen. The hybridization signal was discontinuous in the intima, MCP-1 mRNA expressed where fibroblast and monocyte-like cells assembled. One ruptured aneurysm had no signal because there were no cells only glass-like fiber. Mural thrombus showed upregulated hybridization signal in the cytoplasm of fibroblasts, phlogocytes and endotheliocytes of its micrangium.

CONCLUSIONThe pathological representation of the ruptured and unruptured aneurysms and the upregulated expresion of MCP-1 in the aneurysm wall suggest that the development of aneurysm may be a course of chronic inflammation in which main inflammatory cells are monocyte-like cells.

Inflammatory bowel disease ( IBD) is a chronic disease including Crohn′s disease ( CD) and ulcerative colitis ( UC ) .There are no single diagnostic criteria for IBD .Biomarkers are an objective measurement and evaluation of physiological and pathological processes .And they have been widely used in the diagnosis and prognosis monitoring of IBD patients .

OBJECTIVE: To explore the genetic basis for an infant with neonatal diabetes (NDM) and multiple malformations. METHODS: Genetic variants were detected by next generation sequencing (NGS). Suspected variant was verified by Sanger sequencing. RESULTS: A de novo heterozygous variant, c.1454_1455del(p.K485Rfs), was detected in exon 5 of the GATA6 gene. The variant was undetected in his parents and unreported previously. Bioinformatic analysis predicted the variant to be pathogenic. CONCLUSION The heterozygous variant of c.1454_1455del(p.K485Rfs) of the GATA6 gene probably underlies the disease in this child. Genetic testing can facilitate diagnosis and genetic counseling for NDM.
Abnormalities, Multiple ; Adult ; Diabetes Mellitus/genetics* ; Female ; Genetic Testing ; Heterozygote ; High-Throughput Nucleotide Sequencing ; Humans ; Infant, Newborn ; Male ; Sequence Deletion/genetics*

Hypertension is one of the most important diseases that threaten human health, and has become a global problem. In recent years, the prevalence of hypertension in children has increased significantly year by year, and hypertension in children has a trajectory phenomenon, which is easy to cause cognitive damage and other target organ damage. Therefore, it is necessary to be alert and attached to the relationship between hypertension and cognitive impairment in children. This article reviews the epidemiological characteristics, etiology, diagnostic criteria of hypertension and pathophysiological mechanisms of cognitive impairment caused by hypertension in children.