Objective　To discuss the long-term curative effect of botulinum toxin A(BTXA) for treating spasm of eyelid and facial muscle.Methods　Ninty-two patients with spasm of eyelid and facial muscle were treated with BTXA and observed for a long term.Results　After being injected with BTXA for 1～4 days, the spasm relieved gradually. Sixteen weeks later, the spasm symptoms relapsed. The relapse began mostly from eyelid spasm. The facial spasm's relief time was longer. lt might be up to 24 weeks. After injection, the patients had some slight complications of facial expression numb, ptosis and fissura palpebrae. The above complication symptoms recovered by themselves after two weeks. Conclusion　The curative effect of BTXA on treating spasm of eyelid and facial muscle is reliable.

Purpose　The aim is to develop a synthesis method of the silver carboxymethyl chitosan and to study its bacteriostasis to Staphlococcus aureus(S.aureus),Pseudomonas aeruginusa(P.aeruginusa),Escherichia Coli(E.coli),Klebsiella pneumoniae(K.pneumoniae) and Proteus vulgaris(P.vulgaris).Methods　Chitosan was modified by way of chemistry.The structure analysis of its derivate was analysed by infra-red absorption spectroscopy,using methods of dilution and concavering to study the bacteriostasis to some ordinary bacteria which cause infection in burn.Results　The infra-red spectragram of the derivate showed the chitosan had been modified by chloroacetic acid.When the concentrationl of silver carboxymethyl chitosan was 1.028 mg/ml and the concentration of the five bacteria were 104CFU/ml，the rate of bacteriostasis was 88%、80.2%、75.3% to S.aureus、P.aeruginusa and E.coli respectively.The MIC of silver carboxymethyl chitosan was similar to that of AgNO3 when they act on S.aureus and P.aeruginusa,but the former was lower than later when acting on E.coli.Conclusion　Silver carboxymethyl chitosan could inhibit some bacteria which caused infection in burn.It was a novel pharmaceutical in preventing and curing burn infection.

Objective To investigate the status of knowledge,attitude and behavior of schistosomiasis control of rural resi-dents in Wanjiang River region after a flood,so as to provide the reference for targeted health education. Methods The multi-stage sampling was applied to select the respondents in rural residents in Wanjiang River region,and the self-designed question-naire was used to investigate the current situation of knowledge,attitude and behavior of schistosomiasis prevention and control of the rural residents. Results The total awareness rate of knowledge about the prevention and control of schistosomiasis was 47.92%. The age,education,family income,relatives and friends with medical background,and health education significantly influenced the awareness rate(χ2=12.76,89.19,18.19,50.83 and 92.60 respectively,all P<0.05). The accuracy rates of at-titude and behavior in schistosomiasis control were 62.89%and 52.37%respectively. Conclusion The awareness rate of knowl-edge about the prevention and control of schistosomiasis,and the accuracy rates of attitude and behavior in schistosomiasis con-trol of the rural residents in Wanjiang River region are all inefficient,and therefore,the targeted health education should be strengthened to decrease the risk of schistosomiasis transmission.

Objective To investigate the therapeutic effect of embolizing carotid-cavernous sinus fistula(CCF) by superior ophthalmic vein(SOV) approaches.Design Retrospective case series.Participants 11 patients with CCF diagnosed by digital subtraction angiography(DSA) were failurers of traditional artery approaches.Methods All pathents were treated with embolizing carotid-cavernous sinus fistula by superor ophthalmic vein approaches.Main Outcome Measures visual acuity,exophthalmos,ocular movement,diplopia,conjunctival hyperemia,ocular fundus changes.Results Clinical cure was achieved in all 11 patients during follow-up for 1 week to 3 months.Six patients with symptoms of exophthalmos disappeared and five improved.8 cases with conjunctival hyperemia vanished and 3 cases relieved.The three patients with decreases of 8 visual acuity,among these one patient was normal and two improved.Intracalvarium strepitus and diplopia were all disappeared and ocular movement was normal.Conclusion Embolizing CCF by SOV is safe and effective when performed by a multidisciplinary team.

Background Visual adaptive mechanism of mammalian is close responsible for the development of visual cortex.The various genes with different biological functions in different developing stages of visual cortex participate in regulation of visual development.To investigate the differential expression profiles of various genes in different ages of rat cortex can offer basis and evidence for the study of visual development.Objective Present study aimed to investigate the genes that changed continuously in the postnatal developmental process of rat visual cortex by microarray analysis of visual cortex RNA.Methods Sixty clean SD rats were grouped numbered and randomized into the postnatal day 0 group (P0,n =20),before eye opening group (postnatal day 10,P10,n =15),before the critical period of visual cortex growth group (postnatal day 20,P20,n =15) and the end of development of visual cortex group(postnatal day 45,P45,n=15).The rats were sacrificed at corresponding time point respectively,and the fresh visual cortex were obtained for the extraction of total RNA and microarray analysis.Genes exhibiting changes in expression by≥2.0 folds were further confirmed using real-time PCR(RT-PCR).In order to evaluate the association of differential gene expression with growth,the postnatal stages were paired as 36 groups with the 3 pairs for each target gene (P45/P0,P20/P0,P10/P0).Results Microarray analysis showed that the gene with differential ratio ≥ 2.0 folds in rat visual cortex included Akap7,Asam,Casp3,Cxcr4,Egr1,Ennp2,Fabp7,Gpr88,Inpp5d,Rpsa,Stk32c and Vamp1.Real-time PCR verified that 24 genes form 26 probe sets had the same-phase regulating tendency,including 20 up-regulating probe sets and 6 down-regulating probe sets.The homodromous expressing tendency was seen in Akap7,Asam,Casp3,Cxcr4,Egr1,Ennp2,Fabp7,Inpp5d,Rpsa and Vamp1 genes between microarray analysis and RT-PCR.However,reverse expressions were found in the P45/P0 of Gpr88 and Stk32c genes,showing the up-regulation in the microarray analysis and down-regulation in RT-PCR.The concordant rate of gene expression between microarray analysis and RT-PCR was 94.44％.The expressing genes mainly functioned nervous system development,(metal) ion binding/transport,metabolism,regulation of neuronal synaptic plasticity.Conclusions New relevant candidate genes of age-associated rat visual cortex can be identified by microarray analysis,which provide a clue for the research of visual plasticity.

Objective To explore the effect and mechanism of superficial vein puncture for the first time in obese children auxiliary with venous enhancement.Methods From January 201 5 to July 201 5,1 20 children (3 -7 years old)with obesity were randomly divided into control group and observation group,60 cases in each group.The control group used conventional methods to select limbs superficial vein puncture,the observation group guided by visualizer to select limbs superficial vein puncture.Results The superficial vein puncture success rate of observation group was 96.67% for the first time,the puncture time was (78.95 ±6.68)s;Success rate of control group was 78.33%,the puncture time was (98.44 ±34.1 3)s,the differences between two groups were statistically significant (χ2 =1 0.385,t =21 .787,all P <0.05).Conclusion Obese children with venous visualizer superficial vein punc-ture success rate increased significantly,for the first time the puncture time shortened.

OBJECTIVE: To investigate the expression and significance of miRNA-324-3p and its target gene WNT2B in tissue specimens of nasopharyngeal carcinoma (NPC) specimens. METHOD: qRT-PCR was used to detect the expression of miRNA-324-3p and WNT2B mRNA, and Western blot was applied to assay the expression of WNT2B protein in 39 cases of NPC specimens and 21 cases of non-carcinoma epithelium. The relationship between their expression levels and clinicopathological characteristics and their correlation with clinical pathological parameters was analyzed. RESULT: The expression of miRNA-324-3p was significantly down-regulated decreased but WNT2B mRNA/protein increased obviously in NPC specimens (P < 0.01). A negative correlation between miRNA-324-3p and WNT2B was spotted (P < 0.05). The expression levels of these markers were closely correlated with T stage, clinic stage and cervical lymph node metastasis (P < 0.05). CONCLUSION The loss of miRNA-324-3p and ectopic WNT2B might co-induce the initiation and progression of NPC.
Carcinoma ; Glycoproteins ; genetics ; metabolism ; Humans ; Lymphatic Metastasis ; MicroRNAs ; metabolism ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms ; metabolism ; Neoplasm Proteins ; metabolism ; RNA, Messenger ; metabolism ; Wnt Proteins ; genetics ; metabolism

This study examined the methylation difference in AIRE and RASSF1A between maternal and placental DNA, and the implication of this difference in the identification of free fetal DNA in maternal plasma and in prenatal diagnosis of trisomy 21. Maternal plasma samples were collected from 388 singleton pregnancies, and placental or chorionic villus tissues from 112 of them. Methylation-specific PCR (MSP) and methylation-sensitive restriction enzyme digestion followed by fluorescent quantitative PCR (MSRE + PCR) were employed to detect the maternal-fetal methylation difference in AIRE and RASSF1A. Diagnosis of trisomy 21 was established according to the ratio of fetal-specific AIRE to RASSF1A in maternal plasma. Both methods confirmed that AIRE and RASSF1A were hypomethylated in maternal blood cells but hypermethylated in placental or chorionic villus tissues. Moreover, the differential methylation for each locus could be seen during the whole pregnant period. The positive rates of fetal AIRE and RASSF1A in maternal plasma were found to be 78.1% and 82.1% by MSP and 94.8% and 96.9% by MSRE + PCR. MSRE + PCR was superior to MSP in the identification of fetal-specific hypermethylated sequences (P<0.05). Based on the data from 266 euploidy pregnancies, the 95% reference interval of the fetal AIRE/RASSF1A ratio in maternal plasma was 0.33-1.77, which was taken as the reference value for determining the numbers of fetal chromosome 21 in 102 pregnancies. The accuracy rate in 98 euploidy pregnancies was 96.9% (95/98). Three of the four trisomy 21 pregnancies were confirmed with this method. It was concluded that hypermethylated AIRE and RASSF1A may serve as fetal-specific markers for the identification of fetal DNA in maternal plasma and may be used for noninvasive prenatal diagnosis of trisomy 21.

This study examined the impact of 935MHz phone-simulating electromagnetic radiation on embryo implantation of pregnant mice. Each 7-week-old Kunming (KM) female white mouse was set up with a KM male mouse in a single cage for mating overnight after induction of ovulation. In the first three days of pregnancy, the pregnant mice was exposed to electromagnetic radiation at low-intensity (150 μW/cm(2), ranging from 130 to 200 μW/cm(2), for 2- or 4-h exposure every day), mid-intensity (570 μW/cm(2), ranging from 400 to 700 μW/cm(2), for 2- or 4-h exposure every day) or high-intensity (1400 μW/cm(2), ranging from 1200 to 1500 μW/cm(2), for 2- or 4-h exposure every day), respectively. On the day 4 after gestation (known as the window of murine embryo implantation), the endometrium was collected and the suspension of endometrial glandular cells was made. Laser scanning microscopy was employed to detect the mitochondrial membrane potential and intracellular calcium ion concentration. In high-intensity, 2- and 4-h groups, mitochondrial membrane potential of endometrial glandular cells was significantly lower than that in the normal control group (P<0.05). The calcium ion concentration was increased in low-intensity 2-h group but decreased in high-intensity 4-h group as compared with the normal control group (P<0.05). However, no significant difference was found in mitochondrial membrane potential of endometrial glandular cells between low- or mid-intensity groups and the normal control group, indicating stronger intensity of the electromagnetic radiation and longer length of the radiation are required to inflict a remarkable functional and structural damage to mitochondrial membrane. Our data demonstrated that electromagnetic radiation with a 935-MHz phone for 4 h conspicuously decreased mitochondrial membrane potential and lowered the calcium ion concentration of endometrial glandular cells. It is suggested that high-intensity electromagnetic radiation is very likely to induce the death of embryonic cells and decrease the chance of their implantation, thereby posing a high risk to pregnancy.

Objective To investigate the role of adenosine kinase (ADK) small interfering RNA (siRNA) modified corneal endothelial cells on the proliferation and secretion of regulatory T cells (Treg cells).Methods The experiment was divided into transfected group and non-transfected group,FAM-ADK siRNA was transfected into corneal endothelial cells with Lipofectamine 3000 in transfected group,and the average mass concentration of adenosine was detected by high performance liquid chromatography in both groups.The experiment was divided into control group (transfected with control siRNA),rapamycin group (added 10 ng/ml rapamycin to culture supernatant after transfected with control siRNA) and ADK siRNA group (added 10 ng/ml rapamycin to culture supernatant after transfected with ADK siRNA).TUNEL staining was used to detect the proportion of apoptosis.Flow cytometry was used to detect the expressions of intercellular adhesion molecule-1 (ICAM-1),vascular cell adhesion molecule-1 (VCAM-1) and E-selectin in corneal endothelial cells.The cells were divided into single cultured group (lymphocytes were cultured separately),co-culture group (lymphocytes were co-cultured with corneal endothelial cells) and ADK siRNA group (lymphocytes were co-cultured with ADK siRNA transfected corneal endothelial cells),and the proportion of CD4+ CD25+ Foxp3 + Treg cells was detected by flow cytometry,and the content of interleukin-10 (IL-10) and transforming growth factor-β (TGF-β) in the supernatant was detected by ELISA in the three groups.The access and use of clinical specimens was approved by Ethic Committee of Tianjing Taida Hospital (TD201705140901).Results Flow cytometry showed that 95.1％ of the endothelial cells expressed siRNA.The average mass concentration of adenosine in the supernatant of the transfection group was significantly higher than that in the control group ([38.020±6.658] ng/ml vs.[1.663 ±0.581] ng/ml) (t =5.437,P =0.006).TUNEL staining showed that the average apoptotic cell proportion of corneal endothelial cells in the rapamycin group was significantly higher than that in the control group,and the average apoptotic cell proportion of corneal endothelial cells in the ADK siRNA group was significantly lower than that in the rapamycin group,with significant differences between them (t =3.763,P =0.020;t =4.405,P =0.012).Flow cytometry showed that the average fluorescence intensities of ICAM-1,VCAM-1 and E-selectin in the ADK siRNA group was weaker than that in the control group (4.060± 1.179 vs.11.600±2.427,3.600 ± 1.234 vs.11.030 ± 2.291,5.223 ± 1.734 vs.24.270 ± 4.332),with significant differences between them (t =2.794,P =0.049;t =2.857,P =0.046;t =4.081,P =0.015).The proportions of Treg cells in the single cultured group,co-culture group and ADK siRNA group was significantly different (F =12.890,P =0.007),and the proportion of Treg cells in the ADK siRNA group was significantly lower than that in the co-culture group (t =3.650,P =0.022).ELISA assay showed that,the contents of IL-10 and TGF-β in the supernatant in the single cultured group,co-culture group and ADK siRNA group were significantly different (F =20.960,P =0.003;F =27.320,P=0.001),and the content of IL-10 and TGF-β in the supernatant in the ADK siRNA group was significantly higher than that in the co-culture group,respectively (t =4.492,P =0.011;t =5.280,P =0.006).Conclusions ADK siRNA modified corneal endothelial cells can induce Treg cells to proliferate and secrete IL-10 and TGF-β,which provides a new method for induction of corneal allograft immune tolerance.