Child, Preschool ; Female ; Home Care Services ; Humans ; Infant ; Male ; Positive-Pressure Respiration ; instrumentation ; methods ; Risk Factors ; Sleep Apnea, Obstructive ; prevention & control ; therapy ; Telemedicine ; instrumentation ; methods

Background To optimize the culture method of human retinal microvascular endothelial cells is very important for the study of retinal angiogenesis disease.Human retinal microvascular endothelial cells have been successfully cultured in previous studies,but further improvement of the culture method to harvest higher yields and purity cells is still needed.Objective This study was to design a modified method to isolate and purify human retinal microvascular endothelial cells much easily and quickly,and to compare the expression of specific markers of vascular endothelial cells,factor Ⅷ and CD31/CD34 in the cells.Methods The use of human donor eyeballs was approved by the Ethic Commission of Zhongshan Ophthalmic Center of Sun Yat-sen University.The retina tissue from healthy donor was isolated and digested by the two-step digestion method with 2％ trypsin and 0.133％ collagenase Ⅳ.Human retinal microvascular endothelial cells were collected and plated in 60 mm dishes coated by 0.1％ fibronectin and cultured in endothelial cell-specialized medium supplemented with 10％ fetal bovine serum,0.3 mg/L β-endothelial cell growth factor (ECGF) and 100 ng/L sodium heparin.During the culturing,the growth situation of the cells was monitored by morphological observation,and immunohistochemical staining was performed to probe vascular endothelial cell-specific membrane protein CD31,CD34 and factor Ⅷ for identification of the cell purity.Results Human retinal microvascular endothelial cells were isolated successfully from the retina by the twostep digestion method.The primary cultured cells adhered to well 72 hours later and achieved confluence with the typical cobblestone appearance 9 to 10 days after cultured.The cells exhibited the blue nuclei and reddish cytoplasm by regular haematoxylin and eosin stain and showed a strong positive response for CD31,CD34 and factor Ⅷ by immunohistochemistry.The positive dye of CD31 and CD34 was lower than Ⅷ factor in both endothelial cells.Conclusions Modified culture method of human retinal microvascular endothelial cells can improve cell culture result and purify target cells.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Squamous Cell ; metabolism ; pathology ; radiotherapy ; Esophageal Neoplasms ; metabolism ; pathology ; radiotherapy ; Female ; Follow-Up Studies ; Gene Expression Regulation, Neoplastic ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Inhibitor of Apoptosis Proteins ; Lymphatic Metastasis ; Male ; Microtubule-Associated Proteins ; metabolism ; Middle Aged ; Neoplasm Staging ; Paraffin Embedding ; Survival Rate ; Vascular Endothelial Growth Factor A ; metabolism

Objective To describe the clinical, neuroimaging and genetic profiles of amyotrophic lateral sclerosis with frontotemporal lobe degeneration ( ALS-FTLD).Methods From August 2011 to May 2015, patients with FTLD or other types of neurodegenerative dementia were physically examined in detail and electromyography was performed to those with suspected dysarthria, limb atrophy or weakness.Cognitive and behavioral screenings were performed to all ALS patients.Patients with ALS-FTLD entered further analysis of neuroimaging and genetics.Results Among the 8 patients diagnosed as ALS-FTLD, 4 patients began with personality change or amnesia, while diseases in the remaining 4 cases began with limb weakness or dysarthria.Dementia type of 7 cases was behavioral variant FTLD ( bvFTD) and 1 case was diagnosed as semantic dementia.Electromyography of all the 8 patients showed diffuse neurogenic changes.Constructional neuroimaging of 6 patients showed cerebral atrophy predominantly in frontal and temporal lobes.Fluorodeoxyglucose-positron emission tomography was conducted in 5 patients, indicating hypometabolism mainly in frontal and ( or) temporal lobes.NeuroQ analysis revealed that bilateral frontal lobes were the most hypometabolic areas for ALS-FTLD.Among 4 patients who underwent genetic screening, 1 patient was C9ORF72 mutation carrier.Conclusions bvFTD is the major type of dementia in the context of ALS.Metabolic neuroimaging could assist accurate diagnosis, and it reveals that bilateral frontal lobes are the most hypometabolic areas for ALS-FTLD.C9ORF72 gene mutation is an important pathogenic mutation for ALS-FTLD, although it is rare in Chinese population.

OBJECTIVE: To study the association of interleukin-10 (IL-10) -1082A/G, -819C/T, and -592C/A polymorphisms with IL-10 level and the severity of enterovirus 71 (EV71) infection in children. METHODS: A total of 137 children with hand-foot-mouth disease due to EV71 infection were enrolled as EV71 infection group, which was further divided into mild group with 91 children and severe group with 46 children, and 122 healthy children who underwent physical examination were enrolled as healthy control group. Related clinical data were collected. ELISA was used to measure the serum level of IL-10, and polymerase chain reaction-restriction fragment length polymorphism was used to analyze IL-10 -1082A/G, -819C/T and -592C/A polymorphisms. RESULTS: Compared with the healthy control group, the children with EV71 infection had significantly higher frequency of -1082 AA genotype and A allele (P<0.05). Among the children with EV71 infection, the severe group had significantly higher frequency of -1082 AA genotype and A allele than the mild group (P<0.05), while there was no significant difference in the distribution of IL-10 -819C/T and IL-10 -592C/A polymorphisms between the two groups (P>0.05). The severe group had a significantly higher serum level of IL-10 than the mild group and the healthy control group. IL-10 -1082 AA genotype, -819 TT genotype, and -592 AA genotype were associated with the low expression of IL-10 (P<0.05). As for haplotype, the EV71 infection group had a significantly lower frequency of GCC haplotype than the healthy control group (P<0.05). In the severe group, the children with ATA haplotype had a significantly lower IL-10 level than those with other haplotypes, and the children with GCC haplotype had a significantly higher IL-10 level than those with other haplotypes (P<0.05). There was no significant difference in IL-10 level between children with different haplotypes in the mild group and the healthy control group (P>0.05). CONCLUSIONS IL-10 gene polymorphisms are associated with IL-10 expression and the severity of EV71 infection in children.
Child ; Enterovirus A, Human ; Enterovirus Infections ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Haplotypes ; Humans ; Interleukin-10 ; genetics ; Polymorphism, Single Nucleotide

Objective: To observe the clinical efficacy of mind-refreshing and balance-restoring needling method combined with Frenkel exercises in treating ataxia after cerebral stroke. Methods: The recruited 120 patients were randomized into an observation group and a control group, with 60 cases in each group. The control group was intervened by mind-refreshing and balance-restoring needling method, while the observation group was given additional lower-limb Frenkel exercises. Before and after treatment and at the follow-up, the ataxic lower-limb function was scored using Berg balance scale (BBS) and international cooperative ataxia rating scale (ICARS), and Barthel index (BI) was adopted to score the activities of daily living (ADL). Results: After treatment, the markedly effective rate was 70.2％ and the total effective rate was 96.5％ in the observation group, versus 39.7％ and 87.9％ in the control group, and the differences in the markedly effective rate and the total effective rate were statistically significant (P<0.01, P<0.05). The intra-group comparisons showed that the BBS, ICARS and BI scores after treatment and at the follow-up were significantly different from those before treatment in both groups (all P<0.01).There were significant differences in the BBS score between the two groups after treatment and at the follow-up (P<0.05, P<0.01); the between-group differences in the ICARS and BI scores were statistically insignificant after treatment (both P>0.05), while the between-group differences in the ICARS and BI scores were statistically significant at the follow-up (both P<0.05). The interaction effects between the scoring time of BBS and BI and the group factor were statistically significant (P<0.01, P<0.05). Conclusion: Mind-refreshing and balance-restoring needling can effectively improve the lower-limb ataxic symptoms and ADL after stroke; when combined with Fenkel exercises, this needling method can produce more significant efficacy.

Objective To investigate the differences of blood glycosylated hemoglobin (HbA1c) levels between the patients with acute cerebral infarction and healthy controls, and explore the relation between HbA1c level and both neurological deficits scores and imaging of lesions in number. Methods One hundred and eighty-six patients with acute cerebral infarction within 1 week were performed neurological deficits scales after the admission; the HbA1c level was measured within 24 h admission; brain MRI scan was performed on these patients 48 h after onset or stabilization. Glucose tolerance test was taken at the rehabilitation of infarction (except for having a clear history of diabetes before). At the same time, 160 healthy controls were checked on the level of HbA1c and taken the glucose tolerance test. The differences of blood HbA1c levels between the patients with acute cerebral infarction and healthy controls were investigated; and the relation between HbA1c level and both neurological deficits scores and imaging of lesions in number was explored. Results The HbA1c level in patients with acute cerebral infarction (6.982%±1.803%) was significantly higher than that in the controls (5.128%±0.592%, P<0.05). The level of HbA1c in patients with cerebral infarction and the neurological deficits scores were positively correlated (r=0.760, P<0.05). The level of HbA1c in patients with 2 lesions (6.635%±0.427%) was obviously higher than that in patients with 1 lesion (5.803%±0.307%, P<0.05); The level of HbA1c in patients with 3 or more lesions (8.571%±0.519%) was obviously higher than that in patients with 1 or 2 lesions (P<0.05). Conclusion Diabetes is a major risk factor for cerebral infarction. High HbA1c level might cause a series of cerebrovascular diseases, thus it is an important factor in the happening of cerebral infarction and HbA1c level is an important indicator of the early assessment of the severity of the diseases. The incidence of cerebral infarction can be decreased by controlling hyperglycaemia, lowering the HbA1c levels.

ObjectiveTo evaluate the curative effect difference between Tong's anterior mesh suspension and modified tension-free vaginal tape-obturator (TVT-O). Methods75 SUI cases were random divided into two groups: Group A (35 cases) were accepted Tong's anterior mesh suspension and group B (40cases) were accepted modified tension-free vaginal tape- obturator. Curative effects, operation safety,and complications and so on, were recorded and compared. ResultsThere had no statistical differences between these two groups on operation time, bleeding lose, operation injury, immediate postoperative urinary retention andcure rates, but the operation expense of group A was cheaper than group B [ (980. 74 ±212.45)yuan vs (2879.06 ±467. 13)yuan , P ＜0.05). ConclusionThe curative effect between two methods were similar. It's a little complicated and fit to be popularized in large hospital to modified tensionfree vaginal tape-obturator. Tong's anterior mesh suspension was performed under completely direct vision,not through pelvic cavity, and it was an economic, convenient and easy way, which not only fit to be popularized in basic hospital, but also a surgical remedial measure to the failures who accepted these operations,such as TVT, SPARC, IVS, MONARC, TOT, TVT-O, etc.

Objective To investigate the polymorphism of human cytomegalovirus (HCMV) UL144 gene of the low passage clinical isolates in Guangzhou and explore the role of UL144 gene in HCMV pathogenicity. Methods The clinical isolates of HCMV were obtained from the urine sample collected from those infants with intra-uterus HCMV infection in Guangzhou. The virus genome DNA was extracted. According to the genome sequence of Toledo, primers for UL144 gene were designed and used to amplify the complete open reading frames (ORF) of the UL144 gene in our 3 different clinical isolates. These ORFs of the UL144 gene were cloned into pMD18-T vector and their sequences were confirmed by sequencing. Bioinformatics methods were used subsequently to analyze the polymorphisms of these genes in different stains. Results Three HCMV low passage clinical isolates were successfully isolated, named D2, D3 and D52. As shown by PCR, all of these three strains contained UL144 ORF region. Three complete ORFs were amplified in total and their sequences were submitted to GenBank (Accession No.: DQ180368, DQ180382 and DQ180355). In D2, D3 and D52 isolates, their UL144 ORFs consisted of 531 nucleotides. DNA sequences were quite conservative,all variability were base substitution, and the amino acid sequences were high conservative, the rate of amino acid variability was 1.1%. There were no additional or deleted sites of posttranslational modification of UL144 protein in all clinical isolates. There were some differences in the secondary structure among different isolates. The isoelectric point of UL144 protein of all clinical isolates was 8.97. Conclusions All DNA and deduced amino acid sequences of UL144 gene share great similarity among Guangzhou HCMV clinical strains regardless of their polymorphism. It implies that maybe UL144 gene plays an important role in congenital infection.

OBJECTIVETo test whether the tumor vessel density (TVD) from the enhanced spiral CT can preoperatively predict nodal status and distant metastasis of colorectal cancer.

METHODSForty cases of colorectal cancer patients who received surgical treatment were included in this study. The three dimensional tumor vessels were reconstructed by an enhanced CT 64-slice spiral CT and its AW4.4 image processing platform. The TVD was measured by the 1000 high-resolution color graphics pathological analysis system. The TVD level was compared between different tumor size, classification, and TNM stage. The postoperative pathological staging was taken as golden standard.

RESULTSThe sensitivity, specificity and accuracy for direct prediction of lymph node metastasis by the enhanced CT 64-slice spiral CT was 74.1%(20/27), 53.8%(7/13) and 67.5%(27/40) respectively. The TVD from the reconstructed three dimensional tumor vessels in the group with lymph node metastasis was significantly higher than that without metastasis(0.070±0.046 vs. 0.037±0.013, P<0.05). The TVD in the distant metastasis group was significantly higher than that without distant metastasis (0.130±0.032 vs. 0.049±0.030, P<0.01). No difference of TVD was found between different tumor size, invasion depth, and differentiation type.

CONCLUSIONTVD level from the reconstructed three dimensional tumor vessels can indicate lymph node and distant metastasis of colorectal cancer.