β-Thalassemia is an inheritance anaemia disease due to the defect in β- globin gene. Gene therapy is considered to be the only method which could cure this disease. Adeno-associated virus type 2 (AAV2) has benn gaining more attention as a vector in human gene therapy for its non pathogenic character and broad host range. Although, the efficacy of recombinant AAV2 (rAAV2) in transducing human hematopoietic stem cells has been investigated by researchers, the results were varied from different laboratory. The view was proposed recently that it may be resulted from helper virus in their packaging system. Respecting this, the packaging system without helper virus was used to produce rAAV2. Human early fetal liver hematopoietic cells not only possess many superior peculiarity compared to hematopoietic cells of bone marrow or cord blood, but also the inherent β-globin gene in the cells is not expressed. Studies on the AAV2 transduction of human fetal liver hematopoietic cells and mediated expression of β- globin gene in vivo were performed and the potential role of AAV2 in β-thalassemia gene therapy was analyzed. The rAAV2 containing a normal human β-globin gene (rAAV2-β-globin) without helper virus contamination were produced. The viral titer, purity and the ability of mediating expression of β-globin gene were detected in vitro. Then, human early fetal liver hematopoietic cells were isolated and were further transducted with the rAAV2-β-globin, followed by transplantation into sublethally irradiated BALB/C nude mice to analyze the β-globin gene expression. The results showed that the high titer and purity of rAAV2-β-globin had the ability of mediating β-globin gene expression in vitro. In 8 recipient BALB/C nude mice, the β-globin gene expression were detected in the 2 mice marrow by RT-PCR. The results suggested that rAAV2 could transduce human fetal liver hematopoietic cells and mediate the β-globin gene expression in BALB/C nude mice, meanwhile the expression level of the gene was still rather low. It is necessary to perform further research on AAV2 biology before applying in β-thalassemia gene therapy.

?-Thalassemia is an inheritance anaemia disease due to the defect in?- globin gene. Gene therapy is considered to be the only method which could cure this disease. Adeno-associated virus type 2 (AAV2) has benn gaining more attention as a vector in human gene therapy for its non pathogenic character and broad host range. Although, the efficacy of recombinant AAV2 (rAAV2) in transducing human hematopoietic stem cells has been investigated by researchers, the results were varied from different laboratory. The view was proposed recently that it may be resulted from helper virus in their packaging system. Respecting this, the packaging system without helper virus was used to produce rAAV2. Human early fetal liver hematopoietic cells not only possess many superior peculiarity compared to hematopoietic cells of bone marrow or cord blood, but also the inherent?-globin gene in the cells is not expressed. Studies on the AAV2 transduction of human fetal liver hematopoietic cells and mediated expression of ?- globin gene in vivo were performed and the potential role of AAV2 in?-thalassemia gene therapy was analyzed. The rAAV2 containing a normal human?-globin gene (rAAV2-?-globin) without helper virus contamination were produced. The viral titer, purity and the ability of mediating expression of?-globin gene were detected in vitro. Then, human early fetal liver hematopoietic cells were isolated and were further transducted with the rAAV2-?-globin, followed by transplantation into sublethally irradiated BALB/C nude mice to analyze the?-globin gene expression. The results showed that the high titer and purity of rAAV2-?-globin had the ability of mediating ?-globin gene expression in vitro. In 8 recipient BALB/C nude mice, the ?-globin gene expression were detected in the 2 mice marrow by RT-PCR. The results suggested that rAAV2 could transduce human fetal liver hematopoietic cells and mediate the ?-globin gene expression in BALB/C nude mice, meanwhile the expression level of the gene was still rather low. It is necessary to perform further research on AAV2 biology before applying in ?-thalassemia gene therapy.

Objective To evaluate the relationship between the existence of DME and the SFCT in DR pa-tients. Methods We collected 70 cases which were clinical diabetic involving 122 eyes. They are 36 men with 64 eyes and 34 women with 58 eyes. The average age is 56.7 ± 9.7 year-old. All patients had been examined by oph-thalmologic examination. Grouping: patients without diabetic retinopathy (NDR) group; mild-to-moderate NPDR group; severe NPDR group; PDR group. The last three groups were divided into two subgroup groupsrespectively , which were cases with and without significant macular edema (CSME + / CSME). EDI was used to measure the SFCT. SPSS 17.0 software was used for statistical analysis. Results The average SFCT of NDR group, mild-to-moderate NPDR group, severe NPDR group and the PDR group respectively were (282.94 ± 104.21)μm, (313.62 ± 94.40)μm, (382.44 ± 76.91)μm, (335.00 ± 73.82)μm. Compared with the NDR group, SFCT was thicker than the other three groups, and difference was statistically significant (F = 2.786, P = 2.786). There were no statisti-cally significant difference of SFCT between the mild-to-moderate NPDR/CSME - group and mild-to-moderate NPDR/CSME + group, severe NPDR/CSME - group and severe NPDR/CSME + group PDR/CSME-group, PDR/CSME + group. Conclusions The SFCT of DR patients could potentially be thickenedas disease became serious. When considered the same degree DR, there is no obvious difference of SFCT between patients with CSME and without CSME.

Objective To study the expression and significance of bcl-2 protein in esophageal squamous cell cancer and the surrounding tissues.Methods EnVision method was used to analyze the expression of bcl-2 protein in tissues of esophageal squamous cell cancer and the surrounding tissues from 62 patients.Results Expressions rates of bcl-2 protein were 80.3 ％ (49/61),45.9 ％ (28/61) and 67.7 ％ (20/62) in simple hyperplasia,high grade intraepithelial neoplasia and squamous cell carcinoma tissues,respectively,but 3.3 ％ (1/30) in normal mucosa tissue.There were significant differences between normal esophageal mucosa group and other groups (x2 =54.437,P ＜ 0.01).The expression of bcl-2 had no differentiation in tissue differentiation grade and degree of invasion of carcinoma (x2 =0.219,x2 =5.878,P ＞ 0.05).But it had significant relationship between the expression of bcl-2 and lymph node metastasis (x2 =4.120,P ＜ 0.05).Conclusion bcl-2 may predicting the occurrence of esophageal squamous cell cancer in early stage,and may be regarded as an useful index for prognosis.

Objective To study the effect and mechanism of electroaeupuncture at acupoint Guanyuan on expression of brain-derived neurotrophic factor (BDNF) and tropomyosine receptor kinase B (TrkB) in spinal cord and excitability of detrusor of rat with urinary retention after spinal cord injury. Methods The model of urinary retention after spinal cord injury was established by using weight drop method. Eighty female Sprague-Dawley rats were randomly divided into four groups, namely, a normal, a sham, a model and an electroacupuncture (EA) group, with 20 rats in each group. The EA group was given electroacupuncture at aeupoint Guanyuau once a day for 10 days in total, while the other groups was given no treatment. After 10 days, the expression level of BDNF and TrkB in spinal cord of injury site was measured by immunohistochemistry staining, and excitability of detrusor was measured by using the muscle strips in vitro. Results It was revealed that, with regard to the expression level of BDNF and TrkB in spinal cord,the model group exceeded the normal and sham group, the EA group significantly exceeded the other groups ( P < 0. 05 ). With regard to the excitability of detrusor, the model group was significant-ly lower than the normal and sham group. The EA group significantly exceeded the model group ( P < 0.05 ). Conclusion Electroacupuncture at acupoint Guanyuan can raise the expression level of BDNF and TrkB in spinal cord and increase the excitability of detrusor in rats with urinary retention after spinal cord injury.

Objective:To investigate the clinical and genetic mutation characteristics of a pedigree with familial exudative vitreoretinopathy (FEVR) associated with a LRP5 gene mutation. Methods:A pedigree investigation was performed in a two-generation Chinese Han family with FEVR, which was diagnosed in The First Affiliated Hospital of Xi'an Jiaotong University.Three family members, the proband and his parents, underwent ophthalmic examination, including visual acuity, intraocular pressure, slit-lamp microscopy, fundoscopy and wild-field fundus fluorescein angiography (FFA), to clinically characterize the FEVR phenotype.Peripheral blood of the families were collected for high-throughput sequencing and bioinformatics analysis to identify the pathogenic gene.Sanger sequencing verification was conducted for the detected mutation.The pathogenicity of identified mutations was analyzed according to the guidelines of the American Association of Medical Genetics (ACMG) with software such as Mutation Taster, Polyphen-2, PROVEN and REVEL.This study adhered to the Declaration of Helsinki and was approved by an Ethics Committee of The First Affiliated Hospital of Xi'an Jiaotong University (No.2017-740). Written informed consent was obtained from each subject.Results:The proband was a 27-year-old male.Uncorrected visual acuity (UCVA) was 1.0 for his right eye and 1.2 for his left eye.Fundoscopy showed vascular tortuosity and vasodilation in the temporal peripheral retina of both eyes.FFA indicated that hairbrush-like vasculature and non-perfusion lesion in peripheral retina.The best-corrected visual acuity (BCVA) of the proband's mother, a 51-year old female, was 1.0 for the both eyes.Temporal retinal vascular tortuosity in her left eye was found with fluorescein leakage detected by FFA.The BCVA of the proband's father, a 56-year-old male, was 1.0 for both eyes.Leopard fundus and optic disc atrophy were visible.FFA result was normal.The result of genetic test showed that there were two novel gene mutations in the family with LRP5 gene c. 4110T>G(p.Cys1370Trp) and FSCN2 gene c. 1495G>A(p.Gly499Ser). According to guidelines of ACMG, LPR5 c. 4110T>G was a mutation with uncertain significance.Multiple software, such as MutationTaster, Polyphen-2, PROVEN and REVEL predicted that LRP5 c. 4110T>G mutation might cause detrimental effects on genes or gene products.REVEL scale was 0.93, which indicated that it might be a pathogenic variant. Conclusions:LRP5 gene c. 4110T>G(p.Cys1370Trp) may be a novel mutation for FEVR, which enriches the mutation spectrum of LRP5 gene.

OBJECTIVE:To study the chemical constituents of the ethyl acetate extraction portion of Polygonum bistorta,pro-vide scientific basis for the basic research of its effective substances. METHODS:Taking the ethyl acetate extraction portion of P. bistorta 95％ ethanol extract,positive phase and reversed phase column chromatography were used for repeated isolation and purifi-cation,and the structures of the purified compounds were identified on the basis of physicochemical properties or spectral data. RE-SULTS:11 monomer compounds were isolated and identified as β-sitosterol(1),dihydromyricetin(2),quercetin(3),kaempferol (4),epicatechin(5),chlorogenic acid(6),rutin(7),gallic acid(8),1,2,3,4-tetrahydro-8-hydroxy-4-isopropyl-1-methylnaph-thalene-6-carboxylic acid (9),protocatechuic acid (10) and ellagic acid (11). CONCLUSIONS:The literature is confirmed that compound 2 is isolated from P. bistorta for the first time and compound 9 is isolated from Polygonum for the first time. The related research results have provided experimental references for further clarifying the effective substances of P. bistorta.

Objective To compare the risk factors of breast cancer in Ningxia Hui and Han women,and to provide evidence for the prevention and control of breast cancer.Methods The female patients of breast cancer treated at the Affiliated Hospital of Ningxia Medical University between May 2013 and July 2014 were chosen for case study,while other patients treated at the same hospital and during the same period who did not have breast cancer were selected as a control. An epidemiological survey was conducted using the same questionnaire among the two groups.The survey involved general demographic information,menstrual history,reproductive history,life habit and family history of cancer.Risk factors of breast cancer in Hui and Han nationalities were analyzed by logistic regression analysis.Results Logistic regression analysis showed that the abortion number(OR =2.631,P =0.028)was a risk factor for the occurrence of breast cancer in women of the Hui nationality,while physical exercise (OR =0.177,P =0.040)was a protective factor.Tumor suffered by immediate family members (OR =4.249,P =0.014),abortion number (OR =1.602,P =0.001 ),the age of the first childbirth (OR =1.253,P =0.001 )and the age of first marriage(OR =1.223,P =0.001 )were the major risk factors while physical exercise (OR =0.422,P =0.001 )was a protective factor against breast cancer in Han nationality. Conclusion The risk factors of breast cancer in the women of Hui and Han nationalities are consistent in terms of the total number of abortions and physical exercise.Compared with the Hui people,the age of first marriage,the age of first child birth,and tumor suffered by immediate family members also play a role in the occurrence of breast cancer in the Han na-tionality.

Objective This study aimed to explore the clinical characteristics and treatment of congenital central hypoventilation syndrome(CCHS),to raise awareness on the CCHS for early diagnosis and treatment.Methods The clinical data of 4 babies with CCHS were analyzed,all of the babies were repeated bruising,hypercapnia,and difficult-to-wean.After the relevant checks,we precluded the primary diseases of heart,lung and neuromuscular dysfunction that lead to low ventilation,and detected the major pathogenic genes,Phox2b of CCHS,refer to literature and diagnostic criteria of CCHS.Results Four babies all had typical clinical features of CCHS:enough ventilation when awake.But when they were asleep,their respiratory rate slowed down,hypoventilation,cyanosis and hypercapnia appeared,and they can not be awaked by hypercapnia and hypoxemia caused by hypoventilation.Genetic testing confirmed the presence of Phox2b mutations.Two infants were supported by non-invasive biphasic positive airway pressure(BiPAP),an infant was discharged home at 3 months of age with non-invasive BiPAP,and an infant was discharged home at 1 month of age.Both of them were monitored and treated at home,and showed normal development.Conclusion For babies who were repeated bruising,hypercapnia,and difficult-to-wean,but no primary diseases of heart,lung and neuromuscular dysfunction that lead to low ventilation,CCHS must be considered.Genetic testing of Phox2b can be used as an important diagnostic tool,and non-invasive BiPAP is one of the efficacious methods in the treatment of CCHS.

Objective To compare the difference between GUSS swallowing evaluation and traditional evaluation as a symbol of pulling out stomach tube.Methods 60 patients with cerebral hemorrhage combined with indwelling tube were divided into two groups,the GUSS group and the traditional group,according to the symbol of pulling out the gastric tube.Each group contained 30 cases.The GUSS score ≥ 15 points was used as the symbol of pulling out the gastric tube in the GUSS group,while being able to eat 200 ml liquid without any trouble in two days was used as the symbol in the traditional group.Then we compared the incidence of gastric tube reinsertion and aspiration pneumonitis between the two groups.Results No gastric tube reinsertion and aspiration pneumonitis occurred in the GUSS group,while 7 cases accepted gastric tube reinsertion and 3 cases diagnosed as aspiration pneumonitis in the traditional group.The incidence of gastric tube reinsertion and aspiration pneumonitis in the GUSS group were obviously lower than those in the traditional group.Conclusions GUSS evaluation score ≥ 15 points could be used as a symbol of pulling out gastric tube.