ObjectiveTo explore the clinical features and causative genes of short stature children with unknown etiology， providing evidence for precise clinical diagnosis and treatment. MethodsThe study recruited children with suspected but undiagnosed short stature from the pediatric endocrinology department in our hospital between January 2018 and August 2022. A retrospective analysis was performed on the clinical manifestations， laboratory test and whole exome sequencing （WES） results. Causative genes were classified and analyzed according to different pathogenic mechanisms. ResultsA total of 48 children （30 boys and 18 girls） were enrolled， aged 7.73 ± 3.97 years， with a height standard deviation score （ HtSDS） of -3.63 ± 1.67. Of the patients， 33 （68.8%） suffered from facial anomalies， 31 （64.6%） from skeletal abnormalities， 26 ［54.2%， 61.5% of whom born small for gestational age （SGA）］ from perinatal abnormalities， 24 ［50.0%， 87.5% of whom with growth hormone （GH） peak concentration below normal］ from endocrine disorders and 21（43.8%） had a family history of short stature. Laboratory tests showed that GH peak concentration following stimulation test was （9.72 ± 7.25） ng/mL， IGF-1 standard deviation score was -0.82 ± 1.42， the difference between bone age and chronological age was -0.93 ± 1.39 years. Of the 25 cases with mutant genes found by WES， 14 （56.0%） had pathogenic mutation， 6 （24.0%） likely pathogenic mutation， and 5 （20.0%） mutation of uncertain significance. Pathogenic and likely pathogenic variants were identified in 14 genes， including 10 affecting intracellular signaling pathways （PTPN11， RAF1， RIT1， ARID1B， ANKRD11， CSNK2A1， SRCAP， CUL7， SMAD4 and FAM111A） and 4 affecting extracellular matrix （ECM） components or functions （ACAN， FBN1， COL10A1 and COMP）. ConclusionsA rare monogenic disease should be considered as the possible etiology for children with severe short stature accompanied by facial anomalies， disproportionate body types， skeletal abnormalities， SGA， GH peak concentration below normal and a family history of short stature. WES played an important role in identifying the monogenic causes of short stature. This study indicated that affecting growth plate cartilage formation through intracellular signaling pathways and ECM components or functions was the main mechanism of causative genes leading to severe short stature in children. Further research may help discover and study new pathogenic variants and gene functions.

Objective To investigate the relationship between serum pregnancy associated plasma protein A (PAAA‐A) ex‐pression and gene polymorphism with the severity of coronary lesions in the patients with coronary heart disease (CHD) .Methods Ninety‐eight patients with CHD in the Nanyang Municipal Central Hospital were selected as the observation group and divided into single vessel lesion group and multiple vessel lesion group according to coronary angiographic results .Ninety‐eight individuals un‐dergoing healthy physical examination were selected as the control group .The venous blood was collected at the visiting hospital in the observation group and at the physical examination in the control group for detecting the serum PAPP‐A protein level by ELISA .PAPP‐A gene and IVS6+ 95 polymorphism were analyzed by adopting polymerase chain reaction restriction fragment length polymorphism(PCR‐RELP) .Results Compared with the control group ,peripheral blood PAPP‐A protein level in the obser‐vation group was significantly increased(P<0 .05) ,moreover the PAPP‐A protein level in the multiple vessel lesion group was sig‐nificantly higher than that of the single vessel lesion group (P<0 .05) .The peripheral blood PAPP‐A level was positively correlated with the severity of CHD .Three genotypes existed in PAPP‐A gene IVS6+95 locus ,including CG heterozygous ,homozygous CC and GG homozygous type .The CC homozygous allele frequency in the patients with multiple vessel lesion was higher than that in the patients with single vessel lesion (P<0 .05) .Conclusion The PAPP‐A protein level and IVS6+95 polymorphism have a close relation with the severity of coronary lesions in patients with CHD .CC genotype may be one of genetic susceptibility gene markers in the patients with CHD .

OBJECTIVE:To prepare dexibuprofen sustained-release pellets,and to analyze the drug release behavior in vitro. METHODS:Centrifugal granulation powder layering-eudragit dispersion coating method was used to prepare dexibuprofen sus-tained-release pellets using 3%HPMC as adhesive agent. The formula of the pellets was optimized by orthogonal test with weight ra-tio of sucrose to dexibuprofen,weight ratio of HPMC to Eudragit NE30D and coating weight as factors,using 1,4 and 10 h accu-mulated release rate (Q) as index. The release of the drug from the pellets was analyzed. RESULTS:The optimized formulation was that the proportion of sucrose to drug was 1:10,the weight ratio of HPMC to Eudragit NE30D was 1.5:1,the increased weight of coating material was 8%. Q1 h,Q4 h and Q10 h of prepared pellets were 21%,57% and 89%,respectively(n=3). The co-rrelation coefficient of zero-order,one-order and Higuchi equation release model were 0.956 6,0.989 9,0.996 5. CONCLUSIONS:Prepared pellets show good sustained-release effect in vitro. Drug release of pellets is more in accordance with Higuchi equation.

Objective To study the influence of residual methylene blue after plasma viral inactivation on the human immune cell function by using the peripheral blood mononuclear cell(PBMC) .Methods PBMC were isolated by adopting the Ficoll‐Hypaque density gradient centrifugation method and co‐cultured for 72 h in presence of specific T cell stimulating factors(Anti‐CD3/28 and Anti‐CD28) ,with or without different concentration of methylene blue .The culture supernatant was collected and detected the cyto‐kines secretion situation by ELISA .After 66 h culture ,CCK‐8 dye was added and continueously cultured for 4-6 h ,the prolifera‐tion was determined at A450 .Results The high‐concentration doses of methylene blue (1 .25 ,2 .5 ,5 μmol/L groups) had signifi‐cantly inhibiting effect on the proliferation of PBMC stimulated by Anti‐CD3/28(P< 0 .01) ,its OD value was decreased from 0 .897 ± 0 .385 to 0 .632 ± 0 .334 ,0 .524 ± 0 .254 and 0 .445 ± 0 .287 respectively ,showing certain dose dependent effect .The high concentrations of methylene blue (1 .25 ,2 .5 ,5 μmol/L groups) could down‐regulate interleukin(IL)‐17a ,IL‐10 and interferon (IFN)‐γ secreted by anti‐CD28 induced PBMC ,moreover showing a dose dependent effect .1 .25 ,2 .5 ,5 μmol/L methylene blue af‐fected the IL‐17a level secreted by PBMC from (406 ± 57)pg/mL descending to (276 ± 38) ,(192 ± 31) ,(134 ± 24)pg/mL respec‐tively ;affected PBMC to secrete IL‐10 ,its level was reduced from (184 ± 15) pg/mL to (132 ± 13) ,(110 ± 12) ,(42 ± 8)pg/mL ;af‐fected PBMC to secrete IFN‐γ,its level was deduced from (4 512 ± 187)pg/mL to (2 876 ± 143) ,(2 234 ± 153) ,(1 988 ± 112)pg/mL respectively .Conclusion High concentrations of methylene blue (≥1 .25 μmol/L ) has the significant inhibiting effect on the proliferation and cytokine secretion functions of PBMC .In other words ,the residual methylene blue concentration in viral inactiva‐tion plasma (≤0 .33 μmol/L) has no obvious effect on the immune function of PBMC ,but whether this concentration of methylene blue having the effect on human pure T cell immune function needs to be further evaluated and studied .

Objective:The effects of IL-13 ( Interleukin-13 ) on SDF-1 ( Stromal cell derived factor 1 ) and EGF ( Epidermal growth factor) expression in fibroblasts co-cultured with breast cancer cells were investigated to explore the mechanism for IL-13 in the development of breast cancer.Methods:The co-culture of human breast cancer cell line MDA-MB-231 with the human skin fibroblast line CCC-ESF-1 ( ESF) was used in vitro and in tumor-burdened nude mice.The effects of IL-13 on SDF-1 and EGF expression in the co-cultured fibroblasts in vitro were analyzed using reverse transcription quantitative polymerase chain reaction ( RT-qPCR ) , flow cytometry and Western blot assay.The proliferation of the co-cultured human breast cancer cells in vitro was detected by Cell counting kit-8(CCK-8).The effects of IL-13 on SDF-1 and EGF expression in the fibroblasts of tumor tissue of tumor-burdened nude mice were analyzed using immunofluorescence and laser confocal microscope, and the tumor volumes were examined.Results: IL-13 could up-regulate SDF-1 and EGF expression in the fibroblasts co-cultured with breast cancer cells in vitro,and promoted the proliferation of the co-cultured breast cancer cells.In tumor-burdened nude mice,IL-13 enhanced SDF-1 and EGF expression of fibroblasts in tumor tissue, and accelerated tumor growth.Conclusion:IL-13 up-regulates SDF-1 and EGF expression of fibroblasts co-cultured with breast cancer cells.The molecular mechanism of promoting effect of IL-13 on breast cancer relates to SDF-1 and EGF of fibroblasts in breast cancer stroma.

Pathology is an important basic course in higher vocational nursing,with a wealth of humanistic education.But in the actual teaching process,the humanistic education is often ignored because of too much basic knowledge teaching.Our teaching and research section used questionnaire to find the gaps between current situation,goal of humanistic education and students' needs.By enhancing teachers' humanistic awareness and discovering the entrance to humanistic education,we targeted to professional ideological education,guided students to pay attention to humanistic education and helped them to cultivate humanistic caring capability,and students all had good evaluation on it.It helped to solve the problem of both teaching and education to students in short time under pressure.

Background Stem cell transplantation represents a promising treatment option for patients suffering from degenerative disorders.Accumulating evidences indicate that mesenchymal stem cells (MSCs) are able to differentiate into retinal pigment epithelial (RPE)-like cells.However,MSCs are difficult to obtain.Human adipose mesenchymal stem cells (ADSCs) are proved to have similar properties to MSCs,but relevant study is less.Objective This study was to assess the feasibility of human ADSCs differentiating into RPE-like cells and the safety of its application in vivo.Methods The third generation of human ADSCs were incubated into 6-well plate,and 100 ng/ml epithelial growth factor,50 μ mol/L taurine and 5×10-7 mol/L retinoic acid were added into the medium 12 hours after cultured to induce the cells,and conventional cultured cells were used as the control group.Induced cells were traced with PKH26,and Pan-cytoke ratin (Pan-CK) monoclonal antibody was used to identify the cells under the fluorescence microscope.Induced RPE-like cell suspension of 1 μl was intravetreally injected in the right eyes of 6 BALB/c mice,and equal volume of PBS was used in the same way in another 6 mice.The animals were sacrificed 1 month after injection,and the retinal morphology was examined by histopathology under the optical microscope.The ultrastructure of retinal ganglion cells (RGCs) was examined by the transmission electron microscope.The use and care of the animals complied with Regulations for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission.Results Cultured human ADSCs grew well with the slender polygone shape.Cell membranes showed the red fluorescence for PKH26 after induced.In addition,Pan-CK was expressed in the cell membranes with the red fluorescence in the induced cells,but the response was absent in the control cells.One month after intravitreal injection,induced cells located on the retinal surface,and the retinal morphology was clear under the optical microscope.No abnormality in RGCs was seen under the transmission electron microscope.Conclusions Human ADSCs can differentiate into RPE-like cells after induction.PKH26 can mark induced cells well.There is no adverse effect of induced cells on retina after intravitreal injection in a short-term duration in mice.

Objective To explore the application effect of teach-learn-do integration teaching mode on the course of ‘Fundamental nursing technique’ teaching. Methods We randomly selected one class from each of two 2011 grade and 2012 grade classes as control group and another two classes as experimental group. The students in the experimental group underwent reform teaching method comparing with traditional teaching method used in the control group. In the experimental classes, we reduced the rate of theory and increased the the practice, used the self-written teach-learn-do material, built up double-teaching team, utilized diverse teaching methods. The students in the control group were taken the learning of theory first and then practice this traditional teaching mode. After the course, the theory and skill assessment had been evaluated and compared between two groups. Results The scores of two classes in the experimental group acquired (90. 46 ± 5. 65) score, (87. 41 ± 5. 37) score better than (88. 46 ± 6. 35), (85. 53 ± 6. 45) of the control group, the skill scores (89. 52 ± 2. 30), (88. 48 ± 2. 48) in the experimental group compared with (88. 61 ± 2. 01), (87. 06 ± 3.66) in the control group (t =2. 327, 2. 983, 2. 296, 3. 412;P <0. 05). Conclusions Teach-learn-do teaching method can effectively resolve the gap between theory and practice, enhance the teaching quality of‘Fundamental nursing technique’ , and is adapt the teaching of ‘Fundamental Nursing Technique’ .

Objective To investigate the expression of neive growth factor (NGF) in the ectopic endometrium in adenomyosis patients,and explore the relationship between NGF expression and innervation or pain scales.Methods From Mar.2009 to Oct.2009,45 adenomyosis patients undergoing hysterectomy in Obstetrics and Gynecology Hospital of Fudan University were enrolled in this study,which were classified into 33 cases in pain group and 12 cases in non-pain group based on symptom.The degree of dysmenoreal,chronic pelvic pain and dyspareunia was evaluated by visual analogue scale,including no pain,mild to moderate pain and severe pain group.In the mean time,26 patients with leiomyoma or cervical intraepithelial neoplasia Ⅲ (CIN Ⅲ)undergoing hysterectomy were defined as control group.Ectopic endometrium from experimental group and eutopic endometrium from control group were collected in the surgery.The expression of NGF was examined by immunohistochemistry.The density of protein gene product (PGP)9.5 positive nerve fibers was detected by immuno-fluorescence.Results The NGF level and the density of PGP 9.5 positive nerve fibers in adenomyosis pain group (0.25 ± 0.08,16 ± 8) were higher than adenomyosis painless (0.19 ± 0.05,P =0.007 ; 11 ± 5,P =0.018) and control group (0.18 ± 0.05,P =0.000;9 ± 4,P =0.000).The NGF level and the density of PGP9.5 positive nerve fibers in severe dysmenorrheal group(0.29 ± 0.07,19 ± 10) were higher than mild to moderate dysmenorrheal (0.22 ± 0.07,P =0.018 ; 13 ± 4,P =0.035) and painless group (0.18 ± 0.05,P =0.000 ; 11 ± 5,P =0.006) of adenomyosis patients.There was no difference of NGF level and the density of PGP 9.5 positive nerve fibers in chronic pelvic pain group and no chronic pelvic pain group of adenomyosis patients,so was dyspareunia group and no dyspareunia group.Conclusion The increased NGF level of adenomyosis nodules and improving innervation might be involved in the mechanism of adenomyosis related pain.

Objective To analyze the change of life expectancy (LE) among residents of Fujian province over the past two decade and to evaluate the impact of age and causes of death on the differences related to LE.Methods Mortality data from Causes of Death Surveillance System of Ministry of Health in Fujian province during 1990-2010 were used to calculate the life expectancy,and Arriaga decomposition method was applied to quantitatively evaluate its changes,based on the age-specific and cause-specific mortality.Results The LE in urban and rural residents of Fujian province had an increase of 5.82 and 11.67 years during the past two decade,respectively,which were higher in urban residents than in rural residents.Although the increment of LE was higher in rural populations than in urban population,the difference of LE between urban and rural was seen narrowed.The contribution rate of children on LE was reducing and the contribution rate of children below 14 years old to LE in rural resident declined from 78.29％ to 31.23％.Middle age and elderly populations had become the major ones that causing the change of LE.The impact of cancer,respiratory diseases and cerebrovascular diseases on LE in urban residents was reducing,while the influence of infectious disease,nervous system disease and cardiac disease was increasing,causing the LE of urban resident increased 1.54,0.67 and 0.49 years,respectively.The impact of respiratory diseases,digestive system diseases,as well as injury and poisoning on LE in the rural residents was reducing,while the effect of cancer,cerebrovascular and cardiac disease increased,causing the LE in the rural resident increased 1.23 years.Different causes of death had different impact on the LE in each age group.Conclusion To reduce the mortality rate in the middle-aged and elderly population and to control the incidence of non-communicable diseases could help to improve the LE in the residents of Fujian province.