Objective To investigate the prevalence of vitamin D deficiency and to examine its relationship with the severity and prognosis in the critically ill children. Methods A total of 83 critically ill children admitted from November 1,2010 to December 9,2010 to Pediatric Intensive Care Unit in Beijing Children's Hospital,Capital Medical University were enrolled in the study. Serum 1,25 - Dihydroxyvitamin D concentration was measured by using an en-zyme - linked immunosorbent assay(ELISA). Anthropometric parameters such as height/ length and weight of the chil-dren were measured. Data collection also included primary disease,Pediatric Critical Illness Score(PCIS),the Pediatric Risk of Mortality Ⅲ(PRISM Ⅲ)score,multiple organ dysfunction syndrome( MODS)rate,mechanical ventilation rate,time of hospital of stay and the 28 - day survival rate. Results There were 32 cases with vitamin D deficiency on admission,vitamin D deficiency rate on admission was 38. 6% ,and there was no statistically significant difference among different primary disease groups(P = 0. 815). Vitamin D deficiency rate of malnutrition group was lower than that of the normal group[60. 0%(12 / 20 cases)vs 40. 0%(8 / 20 cases),χ2 = 5. 989,P = 0. 014]. PCIS scores of those with a normal vitamin D status was higher than those of the vitamin D deficiency group,showing a significant difference [(80. 47 ± 6. 18)scores vs(77. 16 ± 7. 59)scores,P = 0. 022]. PCIS score was positively correlated with the vitamin D level(r = 0. 267,P = 0. 015). There was no statistically significant difference among the PRISM score,MODS rate, mechanical ventilation rate,hospital stay length and the 28th day survival rate between the normal vitamin D group and the vitamin D deficiency group(all P ﹥ 0. 05). Conclusions A high prevalence of vitamin D deficiency is found in the critically ill children. The prevalence of vitamin D deficiency in children with malnutrition is higher. Vitamin D status may be correlated to the severity of the critically ill children,but the association with the prognosis is not obvious.

Objective To evaluate the effect of human umbilical cord mesenchymal stem cells (hUCMSCs) treatment through intramuscular administration on the heart function and angiogenesis of the myocardium in dilated car-diomyopathy (DCM)rats induced by Adriamycin(ADR). Methods One hundred male SD rats were randomly divided into the normal group and the DCM group. Rats in the DCM group were treated with ADR by intraperitoneal injection of 2. 0 mg/ kg dose per week for 8 weeks in order to induce DCM. Sixty modeled surviving rats with DCM were randomly divided equally into 3 groups,and they were treated with hUCMSCs or DMEM by intramuscular injection. Rats in the DMEM group (20 cases)received intramuscular infusion 2 mL DMEM alone;rats in the low - dose group (20 cases) underwent intramuscular infusion of 1 × 106 hUCMSCs/ 2 mL in DMEM;rats in the high dose group (20 cases)under-went intramuscular infusion of 10 × 106 hUCMSCs/ 2 mL in DMEM. Echocardiography and plasma brain natriuretic pep-tide(BNP)were used to assess cardiac function in modeled rats. The morphological changes in myocardial cells were observed by using HE and Masson staining after ADR injection stopped for one week. Four weeks after administration of hUCMSCs,echocardiography was performed to evaluate the cardiac function,and plasma BNP level was detected by en-zyme immunoassay kit. Western blot was used to analyze the expression of vascular endothelial growth factor(VEGF)in myocardium of rats in each group. Myocardial microvessel density was detected by using anti - CD34 monoclonal antibody and transmission electron microscopy (TEM)were performed to observe the ultrastructure of microvessel. Results Left ventricular ejection (LVEF)and left ventricular fractional shortening (LVFS)in the DCM groups [(66. 17 ± 3. 54)％,(31. 33 ± 3. 20)％]were significantly decreased compared to those in the normal group [(77. 25 ± 3. 40)％,(41. 00 ± 2. 94)％],and the differences were statistically significant(t = 10. 620,10. 328,all P < 0. 05);the morphological changes in myocardial cells was observed by using HE and Masson staining. Pit - induced typical his-tological lesion of myocardial tissue was observed in the DCM group,such as congestion,edema,a disorganization of myocytes and focal necrosis and myocardial tissue with wispy,broad collagen fibers predominating in the matrix. Four weeks after administration of hUCMSCs,LVEF in the low dose group or the high dose group were significantly higher compared with those in the DMEM group[(72. 27 ± 2. 44)％ or (70. 92 ± 2. 68)％ vs. (62. 89 ± 2. 54)％],and the differences were statistically significant(t = 2. 145,2. 131,all P < 0. 05);and LVFS were significantly higher compared with that in the DMEM group [(34. 96 ± 2. 08)％ or (33. 49 ± 2. 19)％ vs. (30. 98 ± 2. 22)％],and the differences were statistically significant (t = 2. 491,4. 086,all P < 0. 05). The plasma level of BNP was significantly declined in the hUCMSCs treated rats as compared to those before treatment [low dose group (352. 68 ± 41. 25)ng/ L vs. (202. 68 ± 20. 38)ng/ L,t = 2. 052,P < 0. 05;high dose group (355. 79 ± 48. 32)ng/ L vs. (193. 62 ± 15. 41)ng/ L,t = 2. 074,P < 0. 05]. Quantitative analysis demonstrated that microvessel density was significantly hi-gher in low - dose and high - dose hUCMSCs treated DCM rats than that in the DMEM treated DCM rats [(84. 00 ± 19. 18)/ mm2 or (86. 67 ± 20. 88)/ mm2 vs. (27. 14 ± 13. 97)/ mm2 ,t = 2. 109,2. 101,all P < 0. 05];Western blot test showed that there had high expression of VEGF in myocardium and TEM in the high dose group,and vessel injury in DMEM treated rats were more serious than that of hUCMSCs treated rats. Conclusion It suggests that hUCMSCs in-tramuscular injection may improve heart function and angiogenesis of myocardium in DCM rats induced by adriamycin.

OBJECTIVE: To Study the clinical features of congenital microtia and atresia . To evaluate the methods and results of the same microtia surgery, ear canal and middle ear reconstruction. METHOD: Statistically analysis of the data of the hospitalization microtia 62 ears of 58 cases of patient in our department from January 2005 to October 2010 waw conducted. These patients with congenital ear malformations are associated with aural atresia, ossicular chain abnormalities, severe conduction Deafness. All patients received preoperative temporal bone CT examination and reconstruction, hearing examination. Operation was given in two phases, first operation aim to form a line of ear, ear canal reconstruction, ear reconstruction, the second one aim to line of ear skin graft, cranial angle of the ear reconstruction. The preoperative and postoperative data were retrospectively analyzed. RESULT: The auricle plus external auditory canal, middle ear reconstruction came out with a good shape of the ear and the ear canal in close proximity to the normal population. Most patients' hearing were improved after surgery. CONCLUSION Surgeries of patients with congenital ear malformations and aural atresia should be carefully designed according to the three-dimensional reconstruction of multislice spiral CT reconstruction, which can provide information about surgery approach and middle ear abnormality. The whole ear shape and hearing ear after reconstruction are improved after the surgery.
Adolescent ; Angioplasty ; Congenital Microtia ; surgery ; Ear ; Ear Auricle ; abnormalities ; surgery ; Ear Canal ; abnormalities ; surgery ; Ear, External ; abnormalities ; surgery ; Hearing ; Hearing Loss ; surgery ; Humans ; Reconstructive Surgical Procedures ; methods ; Retrospective Studies ; Skin Transplantation ; Tomography, X-Ray Computed

To construct, express and purify Exendin-4 analogue and detect its biological activity in vivo. Insert gene sequence into fusion partner ofpED plasmid which is helped to purification, entitled the new recombinant plasmid 5 Exendin-4 analogue polypeptide gene and fusion partner gene was linked by acid hydrolysisgene, transformed to E. coli BL21 and the fusion protein was induced by lactose. After acid hydrolysis, the Exendin-4 analogue polypeptide separated from fusion chaperon. Anion charge chromatography were used to further purification. 6 to 8 week-old ICR mice were injected (s.c) with Exendin-4 analogue, blood glucose and plasma insulin level was detected in different period after oral glucose tolerance test. The results show that high expression of inclusion body was induced by lactose, which accounted for 40% of germ proteins, the Exendin-4 analogue was obtained with the purity of 91.8% after being purified by anion charge chromatography. Bioactivity assay showed that the level of blood glucose of mouse which treated with exendin-4 analogue was obviously decreased to normal (P < 0.01), and the level of plasma insulin was increased obviously (P < 0.01).
Animals ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Gene Transfer Techniques ; Hypoglycemic Agents ; metabolism ; pharmacology ; Insulin ; blood ; Male ; Mice ; Mice, Inbred ICR ; Peptides ; genetics ; pharmacology ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; pharmacology ; Venoms ; biosynthesis ; genetics ; pharmacology

Objective To investigate the application and the effects of individualized hollow resin plugs in the prevention of external auditory canal stenosis after canaloplasty of external auditory meatus for microtia patients . Methods The clinical data of 65 patients (68 ears) with microtia and aural atresia hospitalized in our department from January ,2006 to December ,2015 were summarized .Otoplasty of external ears and canaloplasty of external auditory meatus were operated on all patients under general anesthesia .Thigh flap transplantation was lined in the bony canal .One month later ,individualized hollow resin plugs were made and worn for 6 to 12 months ,and fol-lowed up for 1～2 years .Results The shapes of the reconstructed auricles were satisfactory .Sixty-four ears with wide external auditory canal openings were recorded in 68 ears .Preoperative hearing was 70 .45 ± 5 .5 dB HL ,and postoperative hearing was 55 .55 ± 5 .2 dB HL .Their hearing was not affected .External auditory canal restenosis occurred in 4 ears .Conclusion The individualized hollow resin plugs do not affect the hearing in use ,and can effec-tively prevent the restenosis of the external auditory canal after microtia reconstruction .

Objective:To evaluate the effects of human umbilical cord mesenchymal stem cells-derived exosomes (hUCMSCs-ex) injection on cardiac function and myocardial fibrosis in dilated cardiomyopathy (DCM) rats induced by Adriamycin(ADR).Methods:One hundred male SD rats were randomly divided into the normal group (20 rats) and the DCM group (80 rats). The rats in DCM group were treated with ADR by intravenous injection to induce DCM.DCM rats were randomly divided equally into DCM group, low-dose group, medium-dose group and high-dose group which were received intravenous injection 1 mL/kg Dulbecco′s modified eagle medium(DMEM), 20 μg/kg, 100 μg/kg and 250 μg/kg exosomes.After modeling, 10 rats in normal group and 30 rats in DCM group were randomly selected to receive echocardiography to evaluate the cardiac function.After exosomes treatment, 10 rats were randomly selected form each group for echocardiography to evaluate the cardiac function.The morphological changes in myocardial cells were observed by using Masson staining in each group; Western blot detection between groups of rats was used to analyze the expression of myocardial collagen Ⅰ type(COLⅠ), Smad2 and alpha smooth muscle actin (α-SMA).Results:Left ventricular ejection fraction(LVEF) and left ventricular fraction shortening (LVFS)in the DCM group [(64.30±3.51)% and (38.70±2.85)%] were significantly lower than those of the normal group [(78.80±1.52)% and (50.60±1.50)%], and the differences were statistically significant ( t=20.518, 22.311, all P<0.01). The left ventricular end-diastolic diameter(LVEDD) and left ventricular end-systolic diameter (LVESD) [(4.62±0.13) mm and (3.40±0.12) mm] of the DCM group were significantly higher than those of the normal group[(3.29±0.24) mm and (3.16±0.33) mm], and the differences were statistically significant( t=2.854, 3.800, all P<0.01). After exosomes treatment, LVEF[(84.3±2.6)% and (83.4±3.2)%] in the medium-dose and high-dose groups were significantly higher than that in the DCM group [(79.2±2.4)%], and the diffe-rences were statistically significant(all P<0.01). Masson staining found that collagen fibers were less in exosomes treating group than those in the DCM group; Western blot test showed that high-dose exosomes can reduce the expression of α-SMA and Smad2, high-dose and low-dose exosomes can both significantly reduce the expression of COLⅠ. Conclusions:It suggests that exosomes intravenous injection from hUCMSCs-ex can significantly improve myocardial fibrosis in DCM rats induced by ADR and cardiac function.

OBJECTIVE To prep are Legumain enzyme and mitochondrial double-stage targeted harmine （HM） liposome （KA@HM-LPS）and preliminary evaluate its pharmaceutical properties ，in vitro antitumor effect and biocompatibility. METHODS Firstly，the preparation and homogenization methods of KA@HM-LPS was screened ，and prepared liposomes were characterized. Secondly，the serum stability ，in vitro release rate ，hemolysis percentage of KA@HM-LPS and cell survival rate under KA@BLPS were determines respectively. Finally ，the cell surivival rate ，mitochondrial targeting and inhibitory effects on cell migration and invasion of KA@HM-LPS were determined. RESULTS KA@HM-LPS was prepared by the thin-film dispersion method ，with encapsulation efficiency of （90.50 ± 0.62）％ . The extrusion moulding method was selected as homogenization method of KA@HM-LPS. The particle size ，polydispersity index ，and Zeta potential of KA@HM-LPS were （211.40±11.67）nm，0.316± 0.014 and（－14.20±0.49）mV，respectively. In 37 ℃，10％ FBS，the particle size of KA@HM-LPS kept stable after 12 h. In vitro release curve of KA@HM-LPS in 20％ plasma conformed to Weibull distribution and had the property of sustained release. When HM concentration was 160 μg/mL，the hemolysis percentage of KA@HM-LPS was （4.23±0.19）％，which was much lower than that of free HM ，with safety. When the mass concentration of KA@BLPS reaches 400 μg/mL，the survival rate of LO 2 cells was （94.40 ± 6.12）％ ，and the biocompatibility was good. Cell test results in vitro showed that ，inhibitory effect of KA@HM-LPS on liver cancer cells with overexpression of Legumain enzyme （LGMN -SK-Hep-1） was significantly higher than that of normal liver cancer cells SK-Hep- 1； compared with SK-Hep-1，LGMN +-SK-Hep-1 cells had a higher uptake efficiency of the liposome ；KA@HM-LPS could significantly inhibit the migration and invasion of LGMN +- SK-Hep-1 cells. CONCLUSIONS KA@HM-LPS is prepared successfully ，which can effectively inhibit the migration and invasion of liver cancer cells with Legumain enzyme overexpression ，and improve the blood compatibility of HM.

Objective: To explore the utility of pharyngeal pH monitoring which positive standard is Ryan index in diagnosis of laryngopharyngeal reflux disease. Methods: In a retrospective study, clinical data of 590 patients who had symptoms laryngopharyngeal reflux disease from February 2016 to March 2017 were analyzed. All patients were received electronic laryngoscopy, assessment of reflux symptom index(RSI) and reflux finding score(RFS), and pharyngeal pH monitoring. SPSS 19.0 software was used to analyze the date. Results: There were 94 patients whose Ryan index were positive(15.93%). Among the 94 patients, 70 were positive during upright, 12 during supine and 12 during both upright and supine. There were 40 patients(6.78%)with pH decline events related to symptoms, while those Ryan index were normal. There were 536(90.85%), 417(70.68%), 233(39.49%) and 117(19.83%) patients with pH<6.5, pH<6.0, pH<5.5 and pH<5.0 events respectively. The positive rate of RSI, RFS, RSI and RFS, RSI or RFS were 44.24%, 16.78%, 7.12%, 53.90% respectively. The RFS score in Ryan index positive group was higher than that in Ryan index negative group[(8.2±2.4) vs (4.0±2.9), u=5.424, P<0.05], while the RSI score in Ryan index positive group was not statistically different from that in Ryan index negative group[(11.3±6.2) vs (12.7±5.8), t=1.247, P=0.167]. Conclusions Pharyngeal pH monitoring is an objective and non-invasive method which can reflect laryngopharyngeal reflux directly. However, with the Ryan index as a criterion for the diagnosis of laryngopharyngeal reflux disease, partial patients may be missed. Further studies are needed to obtain more accurate and objective laryngopharyngeal pH statistical index for diagnosis of laryngopharyngeal reflux disease.