BACKGROUND:Damage to nasal ciliated epithelial cels can lead to a severe injury in nasal biological function. Compared with other adult stem cels, human umbilical cord blood stem cels have better differentiation potential.
OBJECTIVE: To explore the feasibility of human umbilical cord blood stem cels differentiating into nasal ciliated epithelial cels through in vitro culture and induction techniques.
METHODS:Normal and healthy umbilical cord blood samples were colected to isolate human umbilical cord blood stem cels, folowed by identification and subculturein vitro. Umbilical cord blood stem cels at passage 3 were infected with recombinant adeno-associated virus carrying enhanced green fluorescent protein and cultured using air liquid interface culture method. Thereafter, PCR assay was employed for detecting MUCS expression in cultured stem cels at 1 and 2 weeks after induction, and immunofluorescent staining for FOXJ1 was performed at 3 weeks.
RESULTS AND CONCLUSION:After subculture, passage 3 umbilical cord blood stem cels that could express stem cel surface markers were visible in a uniform shape and had good refraction. After 3 hours of gene transfection, green fluorescence issued from the passage 3 cels were visible, and the cel positive rate was up to 96.2% until 48 hours, indicating good transfection efficiency. RT-PCR findings showed that MUC8 mRNA had no expression in the umbilical cord blood stem cels, but expressed strongly in the nasal ciliated epithelial cels, whose expression was weak at 1 week of culture and increased at 2 weeks. Additionaly, the positive expression of FOXJ1 red fluorescence was observed under the transfection of green fluorescent protein. These results suggest that human umbilical cord blood stem cels could differentiate into nasal epithelial cels under suitable conditions.

Objective To explore the relationship between obese children with benign acanthosis ni-gricans and insulin-resistant and type 2 diabetes mellitus. Methods Levels of glucose, insulin, and glucose/ insulin ratio were measured on fasting blood specimens, and anthropometric parameters including waist/hip ratio, fat mass, body fat percentage and body mass index were examined in 42 obese children with benign acanthosis nigricans, 60 cases of simple obesity and 20 healthy children controls. Glucose tolerance tests were performed in groups of obese children with benign acanthosis nigricans and simple obesity, respectively. Results Two of 42 obese children with benign acanthosis nigricans were diagnosed as type 2 diabetes mellitus. The rate of abnormal glucose tolerance and levels of blood sugar during 60 min and 120 min after glucose tolerance were significantly higher in acanthosis nigricans children than those in simple obesity (P

Objective To observe the polymorphism of nitric oxide synthase(NOS)gene and the changes of nitric oxide(NO)in cerebral infarct.Methods The prospective case-control study was employed.Cases contained 193 subjects with cerebral infarction of internal carotid artery system within 2 weeks.Controls contained 103 subjects which came from the normal health checkup.The polymorphism in intron 4 of endothelial nitric oxide synthase(eNOS)gene were detected.NO content was measured by Griess diazotization assay and NOS activity by enzynatic assay.Results There were 48 subjects with allele a in intron 4 of eNOS gene(eNOS4a)in cases and 12 in controls.The frequencies of eNOS4a in cases was higher than that in controls(χ2=8.86,P=0.003).Multivariate Logistic regression analysis confirmed that eNOS4a was an independent risk factors for cerebral infarction(P=0.032).NO production and NOS activity were 6.04(3.83～11.49)μmol/L,(2.97±1.47)U/ml,respectively in cases,and 6.89(4.64～12.43)μmol/L,(3.16±1.46)U/ml,respectively in controls.NO production in cases were significantly lower than that in controls(P=0.022).There was not differential in NOS activity between these two groups(P=0.517).NO production and NOS activity were 5.07(3.18～7.62)μmol/L,(2.77±1.13)U/ml,respectively in the subjects with eNOS4a,and 6.89(4.64～12.43)μmol/L,(3.12±1.54))U/ml,respectively in the subjects without eNOS4a.NO production in the subjects with eNOS4a were significantly lower than that in the subjects without eNOS4a(P=0.001).The NOS activities were not significantly different in subjects with or without eNOS4a(P=0.100).Conclusion eNOS4a possibly exerted some effects on cerebral infarction by diminishing NO.

Objective To observe the final adult height (FAH) outcome and influencing factors in Turner′s Syndrome(TS) children treated with recombinant human growth hormone ( rhGH ).Methods Thirty TS children treated with rhGH were compared with 16 TS children without rhGH treatment and were followed up to achieve their FAH.Comparisons were made regarding predicted adult height (PAH),height standard deviation score for chronological age( HtSDScA ),height SDS for BA( HtSDSRA ),and growth velocity ( GV ) between rhGH treatment and without treatment groups and between the onset and by the end of rhGH treatment group.The factors determining FAH were also evaluated.Results FAH in rhGH treatment group was obviously improved as compared with untreatment group[ ( 149.5±6.3 vs 142.4±5.2) cm,P＜0.01 ].FAH in treatment group was positively correlated with height standard deviation score for chronological age ( Ht0 SDSCA ),Hto SDS for BA ( Hto SDSBA ),height age ( HA0 ) at preliminary diagnosis,and correlated with duration of rhGH therapy,duration of estrogen-free rhGH therapy,and PAH0SDS at preliminary diagnosis.Stepwise regression analysis indicated that duration of estrogen-free rhGH therapy and PAH0 SDS were the variables with the greatest identified influence on FAH (F =11.56 and F =86.91,P＜ 0.01 ).FAH in the 45,XO group was significantly different from the mosaicism group (45,XO/46,XX ) [ ( 147.2 ± 6.3 vs 153.3±6.4) cm,P =0.038].Conclusion rhGH treatment is efficacious in improving FAH of TS children,but a variability in the magnitude of the response to rhGH is recognized.Duration of estrogen-free rhGH therapy and PAH0SDS are the variables with the greatest identified influence on FAH,and karyotype may be one of the influence factors.rhGH treatment should be initiated as early as possible and sufficient course of estrogen-free rhGH therapy is needed to yield a satisfactory FAH.

Objective To evaluate the effect of dexmedetomidine on autophagy in the hippocampal neurons of rats with traumatic brain injury (TBI).Methods Adult male Sprague-Dawley rats,aged 12-16 weeks,weighing 340-370 g,were randomly divided into 3 groups (n=80 each) using a random number table:sham operation group (group S),traumatic brain injury group (group TBI) and dexmedetomidine group (group Dex).The rats were subjected to a diffuse cortical impact injury caused by a modified weight-drop device to induce TBI.Dexmedetomidine 15 μg/kg was injected intravenously immediately after TBI in Dex group.At 24 and 48 h after TBI,neurological deficit score (NDS) was assessed,Morris water maze test was performed,and brains were removed for detection of brain water content in the brain tissue.At 6,12,24 and 48 h after TBI,the expression of hippocampal LC3]Ⅱ was determined using Western blot analysis.Results Compared with group S,brain water content and NDS were significantly increased at 24 and 48 h after TBI,the escape latency was prolonged,and the expression of hippocampal LC3 Ⅱ was upregulated at 6,12,24 and 48 h after TBI in TBI group.Compared with TBI group,brain water content and NDS were significantly decreased at 24 and 48 h after TBI,the escape latency was shortened,and the expression of hippocampal LC3 Ⅱ was down-regulated at 6,12,24 and 48 h after TBI in Dex group.Conclusion The mechanism by which dexmedetomidine reduces TBI is related to inhibition of autophagy in the hippocampal neurons of rats.

OBJECTIVE: To study the regulatory effect of IL-35 on the balance of Treg/Th17 cells in AR patients. METHOD: In this study, 30 cases were randomly selected from outpatients of otolaryngological department in the second hospital of Hebei Medical university who were diagnosed as AR. Another 20 healthy cases enrolled from physical examination branch of our hospital were control group. The expression level of IL-35 and IL-17 in peripheral blood were detected by using ELISA and defeced CD4+CD25+Foxp3+ T cell and CD4+IL-17+T cell expression level were identified via flow cytometry. RESULT: The expression level of IL-35 in AR group was obviously lower than that in control group, and the difference was a statistically significance (t = -8.145, P < 0.01). The expression level of IL-17 in AR group was obviously higher than that in control group, and the difference was a statistically significance (t = 14.969, P < 0.01). There was a remarkable negative correlation between the IL-35 and IL-17 expression in the serum of AR group (r = -0.773, P < 0.01). The percentage of CD4+CD25+Foxp3+ T cell in CD4+ T cell was significant lower in AR group than that in control group (t = -13.678, P < 0.01). The percentage of CD4+IL-17+ T cell in CD4+ T cell was much higher in AR group than that in control group (t = 5.632, P < 0.01). There was a remarkable negative correlation between the Treg and Th17 expression in the peripheral blood of AR group (r = -0.613, P < 0.01). There was a positive correlation between the expression of CD4+ CD25+Foxp3+ T cell and IL-35. There was a negative correlation between the IL-35 and Th17 in AR group (r = 0. -594, P < 0.01). CONCLUSION The lower expression of IL-35 was related to the incidence of AR, and it was an important cytokines for that. The lower expression of IL-35 may inhibit the proliferation of Treg cells, lead to hyper function of Th17 cells, increase secretion of s IL-17 and result in unbalance of Treg/Th17 cells; these may be the important mechanism of the occurrence of AR, thus regulation of IL-35 may become a new target for the immunological therapy of AR.
Case-Control Studies ; Enzyme-Linked Immunosorbent Assay ; Flow Cytometry ; Humans ; Interleukin-17 ; blood ; Interleukins ; Rhinitis, Allergic ; immunology ; T-Lymphocytes, Regulatory ; immunology ; Th17 Cells ; immunology

Objective To explore the value of amino-terminal propeptide of C-type natriuretic peptide (NTproCNP) in evaluating the efficacy of therapy with recombinant human growth hormone ( rhGH ) in patients with idiopathic short stature (ISS) and isolated growth hormone deficiency ( IGHD ).Methods Forty-eight prepubertal children( IGHD n=25,ISS n=23 ) treated for at least 1 year with rhGH were included.Serum insulin-like growth factor- Ⅰ ( IGF- Ⅰ ) and NTproCNP levels were measured before starting treatment and 6 months later.Twelve months after starting treatment,all patients were assessed and annual growth velocity ( GV ),height standard deviation score ( HTSDS),and gained HTSDS (△HTSDS) were recorded.Results In GHD group,positive relationships between GV and change of IGF- ISDS( △IGF- ISDS ),GV and change of NTproCNP concentrations(△NTproCNP) were found( r=0.407,P=0.044 ;r=0.490,P=0.013 ).GH peak value was also positively associated with IGF- ISDS and NTproCNP before therapy ( r =0.558,P =0.004; r =0.630,P =0.001 ).△IGF- ISDS and △NTproCNP were positively associated after therapy ( r =0.466,P =0.019 ).In ISS group,GV was associated with △NTproCNP ( r=0.845,P＜ 0.01 ).Conclusions NTproCNP is a novel biomarker of growth as its level increases during growth-promoting treatment.Furthermore,IGF- Ⅰ is also valuable in evaluating the efficacy of rhGH therapy in short stature patients.

Objectives To evaluate final adult height(FAH), lipid profile, sexual development, and quality of life in individuals with childhood-onset growth hormone deficiency (CO-GHD) during the transition from childhood to adulthood, to reassess the function of GH-IGF-I axis, and to explore effective managements for different types of GHD in each period. Methods Totally 80 CO-GHD patients were divided into 2 groups; 22 patients with isolated growth hormone deficiency ( IGHD) and 58 patients with multiple pituitary hormone deficiencies (MPHD); 62 male (age ≥18 years) and 18 female ( age ≥ 16 years) patients. The clinical and biochemical parameters, education and occupation, rhGH, and other hormones therapy in the past were followed up. Results rhGH replacement improved FAH of patients with GHD. The incidences of either hyperlipidemia (39.0% , 47.4%) or fatty liver disease (26.8%, 31.6%) showed no statistically significant changes between 2 groups with and without rhGH replacement. Mean value of IGF-I SDS was significantly higher in IGHD group than that in MPHD group (-1.43±0. 31,-3. 01 ±0. 66) ,and also IGFBP3(-2. 10±0. 33,-3. 17±0. 19,all P< 0.05 ). Patients with IGHD had normal sexual development, but the incidence of sexual dysfunction accounted for 79.7% in MPHD group. Conclusions rhGH improves FAH of individuals with CO-GHD. Patients with CO-GHD should be followed during the transition period; GHD patients carry a high risk of metabolic abnormalities in the adulthood; IGHD female can give birth to offsprings; patients with MPHD have gonadotrophin deficiency of varying degrees.

OBJECTIVETo investigate the clinical symptoms and potential mutations in the PLA2G6 gene for a child with infantile neuroaxonal dystrophy.

METHODSClinical data of the patient was collected. The coding regions of PLA2G6 gene was subjected to Sanger sequencing using blood DNA from the patient and her parents.

RESULTSThe patient has presented with psychomotor regression and hypotonia, followed by development of tetraparesis. A novel homozygous mutation G68A in the PLA2G6 gene was found by DNA sequencing, while her parents were both heterozygous carriers.

CONCLUSIONThe psychomotor regression and tetraparesis of the patient was caused by infantile neuroaxonal dystrophy due to a novel homozygous mutation in the PLA2G6 gene, which was inherited from her parents.

Adult ; Base Sequence ; Brain ; diagnostic imaging ; Child, Preschool ; DNA Mutational Analysis ; Female ; Group VI Phospholipases A2 ; genetics ; Homozygote ; Humans ; Magnetic Resonance Imaging ; Male ; Molecular Sequence Data ; Mutation ; Neuroaxonal Dystrophies ; diagnostic imaging ; genetics ; Radiography

Parkinson's disease (PD) is a common degenerative disease of the central nervous system, but no drug has been found to be surely able to protect neurons so far, delay onset or slow progression of the disease. Currently there are a variety of Chinese formulas, single herb medicines, active fractions and monomers showed prophylactic and therapeutic effect on PD animal models. The mechanisms include protection of substantia nigra cells, improvement of neurotransmitter content, anti-oxidation, immune regulation, enhancement of Western medicine efficacy, reduction of side effects, etc. All these mechanisms may play integrated effect and slow disease progression. In particular, Chinese medicine compound may have some advantages in neuroprotective treatment of PD, because a variety of active ingredients can exert multi-links, multi-levels and multi-targets integrated regulation effect on human body. However, the level and standard of Chinese medicine studies on PD animal still need to be improved.
Animals ; Antioxidants ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Humans ; Medicine, Chinese Traditional ; methods ; Neuroprotective Agents ; pharmacology ; Parkinson Disease ; drug therapy ; immunology ; Substantia Nigra ; drug effects ; metabolism