OBJECTIVE: To provide a reference about choosing the methods of isolating exosomes derived from tumor cells including laryngocarcinima Hep-2 cells by comparing advantages and defects of two methods of isolation and extraction exosomes. METHOD: Previously, laryngocarcinoma Hep-2 cells were cultivated massively, then the cells were processed with hot shock in 42 degrees C for 1 h. Sucrose density gradient centrifugation ultrafiltration (method 1): cells culture supernatant 90 ml was gathered, the supernatant was clarified through a 3/0.8 μm small filter to remove impurities and fragments which in larger diameter. Then the filtering fluid was concentrated and purified through sucrose density gradient centrifugation and ultrafiltration, the concentrated fluid was obtained. Exosome Isolation Kit (method 2): cells culture supernatant 4 ml was gathered, the solutions of the kit were added into the supernatant in proper sequence, then filtered by the special column, the concentrated fluid was obtained. Both products are observed by high resolution transmission electron microscopy. RESULT: Both methods could isolate and extract exosomes feasibly. In single high power view of transmission electron microscopy, exosomes of method 1 disperse better, but lower density, and more impurity in background, exosomes of method 2 arrange closer, higher density, and less impurity. CONCLUSION Exosome isolation Kit require less supernatant, cost less time, process procedure briefly, harvest higher yield. It may become a new option of isolating exosomes derived from Laryngocarcinoma Hep-2 cells.
Cell Line, Tumor ; Exosomes ; ultrastructure ; Humans ; Laryngeal Neoplasms ; pathology ; Microscopy, Electron, Transmission

A method of complete acid hydrolysis combined with high performance anion exchange chromatography and pulsed amperometric detection was developed for the monosaccharide composition analysis of arabinoxylan from the seeds of Plantago asiatica L. The parameters including hydrolysis methods, acid types, acid concentration, hydrolysis temperature, hydrolysis time and placement time, which would affect the hydrolysis process, were optimized. The results showed that it would have a better hydrolysis effect for polysaccharide from the seeds of Plantago asiatica L. with 2 mol/L H2 SO4 in an atmospheric oil bath at 120℃for 2 hours. However, the placement time for diluted solution of the hydrolyzed polysaccharide should be less than 6 hours. The polysaccharide was mainly composed of Arabinose (8. 89%) and Xylose (41. 52%) and Galacturonic acid (0. 73%). Glcuronic acid (3. 44%) was detected simultaneously, and there were also trace amounts of Galatose and Glucose. The results were reproducible. Other arabinoxylans from Panicummiliaceum L. shell, Avena sativa L. bran and Hordeum vulgare L. were taken for monosaccharide compositions analysis under the optimal hydrolysis conditions and the analysis results were good. This study would provide a good reference for monosaccharides composition analysis of arabinoxylans from various sources.

OBJECTIVE: To study surgical techniques and clinical applications of the intranasal endoscopic combined middle meatus and expand prelacrimal recess-maxillary ainus approach for orbital fracture treatment. METHOD: A retrospective clinical analysis of 3 patients whose admitted for orbital floor fractures or medial wall fractures operated by the intranasal endoscopic middle meatus with expand prelacrimal recess-maxillary ainus approach surgical treatment was studied, and the treatment effects and the postoperative complications were analyzed. RESULT: All patients had been followed up for 6 to 12 months. All cases of diplopia symptom were disappeared, enophthalmos were totally corrected, no cases of complication were found. CONCLUSION Endonasal endoscopic combined middle meatus and expand prelacrimal recess-maxillary ainus approach for orbital fracture treatment have great and clear view. This approach with less tissue damage and high therapeutic effect makes the cost lower than other methods and complications will be decreased as well, it has a great advantage in the orbital fracture treatment.
Diplopia ; etiology ; therapy ; Endoscopy ; Enophthalmos ; etiology ; therapy ; Humans ; Maxillary Sinus ; surgery ; Nose ; Ophthalmologic Surgical Procedures ; methods ; Orbital Fractures ; complications ; surgery ; Postoperative Complications ; Reconstructive Surgical Procedures ; methods ; Retrospective Studies

Objective To investigate the clinical and molecular genetic changes in a Chinese family with oculopha?ryngeal muscular dystrophy(OPMD). Methods We collected the clinical data of the familial members and blood sam?ples from all available 16 familial members, including the proband. The samples were analyzed using modified poly?merase chain reaction amplification and direct sequence analysis. Results Male OPMD patients initially presented with ptosis, followed by pronunciation difficulty, dysphagia and limb weakness whereas female OPMD patients initially pre?sented with swallowing difficulty. Genetic test revealed the abnormal expansions of the GCG trinucleotide repeat from GCG6 to GCG10 in PABPN1 gene in 10 familial members. Conclusions The genetic test and prenatal diagnosis is the key for the prevention treatment of oculopharyngeal muscular dystrophy. The ptosis of eyelid may be the initial symptom for the male patients of oculopharyngeal muscular dystrophy with (GCG)10 mutation.

Objective: To understand the current situation of infrastructure construction and sanitation practice of school canteens in China and to provide a reference for improving food safety in school canteen and sanitation management level. Methods: Stratified sampling method was adopted to select the surveyed schools. Information regarding school canteen infrastructure and sanitation practice was collected through questionnaires. Chi-square test was used to compare the difference in passing rate on the survey indices among different school types. Results: In environmental sanitation, 91.0% of the school canteen was rated as good and in personal hygiene, 92.8% of the employees were rated as good. The overall health level of urban schools was better than that of township schools, and non-boarding schools was better than that of boarding schools, with statistically significant differences(P<0.01). 67.1% of school canteens provided with independent food outlets while 98.7% of school canteens without independent food stores. The qualified rate of washing facilities in the dining area was 94.7%, and the qualified rate of distance between the canteen and the pollution source was 92.5%. Nearly all the employees held canteen catering service license (99.0%) and the health qualification certificate (99.5%). Most of the school canteens (98.7%) established the safety rules and regulations, including the food retention sample system. Nearly 61.5% schools initiated the principal meal program. Conclusion Sanitation management in the canteens of primary and secondary schools in China is generally satisfactory, with some canteens still fail to meet the standard requirements. More supervision from schools and relevant government departments (market supervision, health and education departments) is needed to provide students with a clean and heathy campus canteen.

Objective: To investigate routine blood test results and secular changes among Tibetan children and adolescents aged from 3 to 19 in the plateau, and to provide the basis for reference range of routine blood test for this population. Methods: A total of 1 568 Tibetan children and adolescents aged from 3 to 19 living in Shigatse, Tibet were selected by cluster random sampling method. Routine blood test results and its secular trends were compared by age and gender. Results: Significantly differences were found in red blood cell(RBC), hemoglobin(HGB), hematocrit (HCT), mean corpuscular volume (MCV), mean corpuscular hemoglobin(MCH), white blood cell(WBC), neutrophil(NEU), neutrophil percentage (NEU%), lymphocyte(LYM), lymphocyte percentage(LYM%)，monocyte percentage(MON%)，eosinophil percentage(EOS%),basophil percentage(BAS%) and platelet(PLT) among the four age groups of 3-5, 6-12, 13-15, and 16-19 years ( F/H =60.22, 179.41, 249.45, 115.03, 74.90, 14.33 , 33.46, 78.90, 49.20, 97.29, 24.45,24.28,42.65,20.10, P <0.05). Among red blood cell indexes, RBC, HGB, HCT,MCH increased with age in boys( F =148.77, 493.04, 623.14, 249.92, P <0.05), but there was no similar trend in girls( F =1.37, 0.15, 2.94, 0.11, P >0.05). HCT showed significant sex differences among the four age groups of 3-5 years, 6-12 years, 13-15 years, and 16-19 years [(41.33±2.31)% vs (41.98±2.40)%; (43.28±2.60)% vs ( 43.75 ±2.36)%; (46.20±3.11)% vs (44.83±2.67)%; (51.10±4.15)% vs (43.61±4.70)%, t =-2.10, -2.88, 3.50, 10.82, P <0.05]. WBC, NEU, NEU%, LYM, LYM%, monocyte(MON), and MON% increased significantly with age in both boys and girls ( P <0.05). From the age of 12 to 13, RBC, HGB and HCT in Tibetan male and female adolescents showed an opposite trend and widened gradually. Conclusion Red blood cell index shows significantly different trends among Tibetan adolescents and children of different ages and genders. Regional nationality, age, gender, and other factors should be considered when developing the reference value range of blood routine index.

To report a case of Noonan-like syndrome with loose anagen hair, and to detect gene mutations in the family. A 3-year-old female patient presented with a special facial appearance, short stature, deep palmar and plantar creases, and dark skin. The scalp hair was sparse and thin, dermoscopy showed black spot sign, single-hair follicular units, hair shafts varying in the diameter, upright hair and new vellus hair, and constriction of hair shafts and nodular hair were occasionally observed. Peripheral blood samples were collected from the proband and her parents, and the genomic DNA was extracted for whole-exome sequencing. A heterozygous missense mutation c.4A>G was identified in exon 2 of the SHOC2 gene in the proband, resulting in the substitution of serine by glycine at amino acid position 2 (p.S2G) . The mutation was not identified in either of her parents. Combined with the clinical phenotype and gene mutation, the diagnosis of Noonan-like syndrome with loose anagen hair was confirmed.

Objective: To investigate the specificity of endogenous metabolic profile in plasma of patients with occupational acute methyl acetate poisoning using non-targeted metabolomics. Methods: A total of six patients with occupational acute methyl acetate poisoning were selected as the poisoning group, while 10 healthy workers without occupational exposure history of chemical hazards in the same industry were selected as the control group using the judgment sampling method. Metabolites in patient plasma of the two groups were detected using ultra-high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry, and non-targeted metabolomics analysis was performed. Principal component analysis and partial least squares discriminant analysis were used to identify differential metabolites and analyze their metabolic pathways. Results: There were significant differences in metabolite profiles in patient plasma between poisoning group and control group. A total of 195 differentially expressed metabolites were screened in plasma of patients in poisoning group, including 119 upregulated and 76 downregulated metabolites. Lipid substances (lipids and lipid-like molecules) accounted for the highest proportion (21.5%). The differential metabolites of poisoning group were related to folate biosynthesis, amino acid metabolism, pyrimidine metabolism, sphingolipid biosynthesis and other metabolic pathways in plasma compared with the control group (all P<0.05). Conclusion: Occupational acute methyl acetate poisoning affects metabolism of the body. The folic acid biosynthesis, amino acid and lipid metabolism and other pathways may be involved in the occurrence and development of poisoning.

Objective: To summarize the skill and experience of transnasal endoscopic operation for retrobulbar lesions. Methods: Seven patients aged from 25 to 67 years old diagnosed as retrobulbar lesions who underwent transnasal endoscopic operation in Department of Otorhinolaryngology Head and Neck Surgery, Xiangya Hospital between January 2013 and October 2016 were retrospectively analyzed. Two males and five females were included in this study. Five patients underwent transnasal endoscopic operation via media rectus-inferior rectus space, with the other 2 cases via media rectus-superior rectus space. Results: Total lesion removal was achieved in 6 of 7 patients, while 1 patient underwent subtotal removal of the lesion. The visual acuity and visual field improved in 3 cases. The pathological examination showed hemangioma(5 cases), bone cyst(1 case) and fibroma(1 case). All patients were followed up for 9 months to 4 years without complications such as eye movement disorder or blindness, except for 1 case with preoperatively proptosis occurred postoperatively transient diplopia. There was no recurrence in 6 patients with total lesion removal, and the patient underwent subtotal removal of fibroma did not undertake operation again. Conclusion Transnasal endoscopic operation for retrobulbar lesions is a minimally invasive, safe and effective operatiiv method, which could be taken via different surgical approaches according to the size and location of the lesion.

OBJECTIVEThe aim of the present study was to assess the association between the polymorphisms of CD36 gene and acute coronary syndrome(ACS).

METHODSGenotypes of CD36 single nucleotide polymorphisms were detected under PCR-RFLP in 522 patients with ACS and 1 215 controls.

RESULTSFor people under Han and Uygur ethnicities, the distribution of genotypes and allele of rs1722505 was significantly different between ACS and the controls(all P < 0.05). For Han population, the frequency of A allele of rs17154181 was significantly lower in ACS group than that in the control group(P = 0.034). Results from logistic regression analysis showed that the AA+AG genotype of rs1722505 was significantly higher in ACS patients than that in controls both between the Hans and the Uygurs(OR = 1.436, 95%CI:1.047-1.970, P = 0.025;OR = 1.589, 95%CI:1.009-2.473, P = 0.046, respectively). For Han people,AA+AG genotype of rs17154181 was significantly lower in ACS patients than that in controls(OR = 0.667, 95% CI:0.494-0.900, P = 0.008).

CONCLUSIONData from the present study suggested that the CD36 gene might serve as a genetic marker of ACS in both Han and Uygur populations.

Acute Coronary Syndrome ; epidemiology ; genetics ; Alleles ; CD36 Antigens ; genetics ; China ; epidemiology ; Ethnic Groups ; genetics ; Gene Frequency ; Genotype ; Humans ; Polymorphism, Single Nucleotide