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1.
Article in English | WPRIM | ID: wpr-785830

ABSTRACT

BACKGROUND AND OBJECTIVES: The exosomes released by mesenchymal stromal cells (MSCs) in classical FBS-containing media have been demonstrated as an alternative, cell-free therapy in various diseases including inflammatory bowel disease (IBD). It has been found that the function of exosomes is affected by culture condition. We previously developed a serum-free, xeno-free and chemically defined medium, and umbilical cord-derived MSCs in this medium retained the immunosuppressive capability.METHODS: In this study, we evaluated the immunosuppressive function of exosomes from MSCs (MSC-Exo) in defined medium and their therapeutic effect on treating colitis.RESULTS AND CONCLUSIONS: In vitro studies indicated that MSC-Exo reduced the concentration of pro-inflammatory cytokines IFN-γ, TNF-α and IL-1β, and increased the secretion of anti-inflammatory cytokines TGF-β1 and IL-10, but no significant change of inhibitory effect on peripheral blood mononuclear cells proliferation was shown. In vivo experimental colitis showed that administration of MSC-Exo was able to significantly ameliorate the disease activity index score, weight loss, colon shortening, and the histological colitis score through up-regulation anti-inflammatory responses and down-regulation of inflammatory responses. Moreover, the use of MSC-Exo (200 μg) led to an improved therapeutic efficacy when compared with MSCs at a dose of 1×10⁶ cells. Our findings indicate that the exosomes from MSCs in defined medium possess a certain degree of immunosuppressive effect in vitro and exhibit a therapeutic capability in a mouse model of DSS-induced colitis through suppressing inflammation mechanism.


Subject(s)
Animals , Colitis , Colon , Cytokines , Down-Regulation , Exosomes , In Vitro Techniques , Inflammation , Inflammatory Bowel Diseases , Interleukin-10 , Mesenchymal Stem Cells , Mice , Up-Regulation , Weight Loss
2.
International Eye Science ; (12): 709-711, 2015.
Article in Chinese | WPRIM | ID: wpr-637240

ABSTRACT

AIM: To explore the relationship between different timing of vitreous surgery and prognosis after the open globe injuries. METHODS:Fifty-eight cases (58 eyes) with open globe injuries were chosen in our hospital from June 2011 to June 2013, and randomly divided into observation group and control group to have vitreous surgery in early phase and normal time respectively. Thirty cases ( 30 eyes ) in the observation group were received vitreous surgery during 3d, and 28 cases (28 eyes) in the control group from one to two weeks. The vitreous surgery of two groups of patients were finished by the author herself. The ocular injury before surgery, the effect of surgery treatment and the incidence of complications after surgery of the two groups of patients were compared. RESULTS: The eye damage situation such as lens rupture, vitreous hemorrhage, retinal detachment and corneal laceration of two groups of patients before surgery were no significant statistical difference ( P >0.05). The postoperative therapeutic effect of observation group was better than the control group (χ2=6. 869, P<0.05 ); the postoperative complications of observation group was obviously lower than the control group (P<0. 05). CONCLUSION: The timing of vitreous surgery of patients with open globe injuries correlate with the prognosis, which prompts that better therapeutic effect can be achieved and the occurrence of complications can be reduced in early vitreous surgery of the patients with open globe injuries.

3.
Chinese Journal of Pediatrics ; (12): 915-919, 2013.
Article in Chinese | WPRIM | ID: wpr-288813

ABSTRACT

<p><b>OBJECTIVE</b>To reveal the molecular genetic pathogenesis of the glycogen storage disease type III (GSDIII) and to provide a prerequisite for prenatal gene diagnosis in future.</p><p><b>METHOD</b>All the coding regions as well as the border areas between exons and introns of the AGL gene and the parental relevant mutation sites were directly sequenced, so that to affirm the origin of the mutation. Then, detected novel heterozygous mutation was confirmed by cloning sequencing. Finally, definite diagnoses of the novel mutation were performed by a series of identification methods, including screening for the 100 normal controls by DHPLC in order to count the mutational frequency, analyze the conservative of the mutant amino acid sequence from 11 kinds of species and comprise the difference of the tertiary structure between the mutant protein and the normal one.</p><p><b>RESULT</b>The patient had compound heterozygous mutations, the c.100C>T (p.R34X) nonsense mutation and c. 1176_1178 del TCA deletion mutation. The p.R34X has been reported abroad, but the 1176_1178 del TCA/p.His392fs mutation is a novel one. The proband's father is heterozygous with the p.R34X mutation while his mother carries the c.1176_1178 del TCA mutation. The result from searching the dbSNP database, HGMD database and papers published in recent years showed that the c.1176_1178 del TCA is a novel mutation, but not an SNP. Conservative analysis results in 11 species indicate that the amino acid of the mutation site is highly conserved in the stage of evolution. Comparison results between the mutant protein and the normal one demonstrate that the deletion mutation results in the obvious variation of the spatial conformation of AGL protein.</p><p><b>CONCLUSION</b>The "c.1176_1178 del TCA (p.392delHis)" mutation is a novel pathogenic mutation. This mutation and the c.100C>T (p.R34X) is the cause that the proband suffer from the GSDIIIa disease. These two mutations are inherited from mother and father respectively. The methods from this paper can be used for further prenatal gene diagnosis.</p>


Subject(s)
Adult , Amino Acid Sequence , Base Sequence , Case-Control Studies , Child, Preschool , DNA Mutational Analysis , Exons , Female , Glycogen Debranching Enzyme System , Chemistry , Genetics , Glycogen Storage Disease Type III , Diagnosis , Genetics , Heterozygote , Humans , Male , Mutation , Pedigree , Polymerase Chain Reaction , Protein Conformation , Sequence Alignment
4.
National Journal of Andrology ; (12): 890-895, 2013.
Article in Chinese | WPRIM | ID: wpr-350817

ABSTRACT

<p><b>OBJECTIVE</b>To study the correlation of azoospermia and severe oligozoospermia with Y chromosome microdeletions, chromosome karyotype and reproductive hormones in male infertility patients.</p><p><b>METHODS</b>We collected semen samples from 63 patients with azoospermia, 49 with severe oligozoospermia and 60 men with normal semen parameters, and determined the incidence of Y chromosome microdeletions, chromosome karyotypes and the levels of reproductive hormones.</p><p><b>RESULTS</b>The incidence rate of Y chromosome microdeletions was 11.11% in the azoospermia and 8.16% in the severe oligozoospermia patients, as compared with 0 in the normal controls (P<0.05). The rate of chromosome abnormalities was 9.52% in the azoospermia group, with statistically significant differences from the severe oligozoospermia and normal control men (both 0) (P<0.05). The levels of FSH and LH were significantly higher in the azoospermia ([20.41 +/- 19.34] IU/L and [11.44 +/- 9.48] IU/L) and the severe oligozoospermia patients ([8.88 +/- 7.04] IU/L and [6.78 +/- 3.85] IU/L) than in the normal males ([3.88 +/- 2.21] IU/L and [4.63 +/- 1.51] IU/L) (P<0.05).</p><p><b>CONCLUSION</b>Examinations of genetics and reproductive hormones are necessary for infertile males with azoospermia and severe oligozoospermia, which may contribute to early diagnosis and treatment.</p>


Subject(s)
Adult , Azoospermia , Genetics , Case-Control Studies , Chromosome Deletion , Chromosomes, Human, Y , Genetics , Hormones , Blood , Humans , Infertility, Male , Karyotype , Karyotyping , Male , Oligospermia , Genetics , Semen , Sex Chromosome Aberrations , Sex Chromosome Disorders of Sex Development , Blood , Genetics , Sperm Count
5.
Article in Chinese | WPRIM | ID: wpr-350691

ABSTRACT

<p><b>OBJECTIVE</b>To establish a method of TLC identification for Dida commonly used in Tibetan medicine from different species.</p><p><b>METHOD</b>With silica gel G as the stationary phase, and chloroform-methanol (40: 1) as mobile phase, oleanolic acid from different species of Dida was separated and identified.</p><p><b>RESULT</b>Oleanolic acid was detected in 70 kinds of Dida derived from the Gentianaceae Swertia, Halenia, Gentianopsis, Lomatogonium, and Saxifragaceae saxifrage, except for the saxifrage, there are some differences among different genera or subjection.</p><p><b>CONCLUSION</b>This TLC method can be used for identification of oleanolic acid in Dida from different species except saxifrage.</p>


Subject(s)
Chromatography, High Pressure Liquid , Chromatography, Thin Layer , Methods , Drugs, Chinese Herbal , Chemistry , Medicine, Tibetan Traditional , Methods , Oleanolic Acid , Chemistry , Species Specificity
6.
Article in Chinese | WPRIM | ID: wpr-733062

ABSTRACT

Objective To discuss the magnetic resonance imaging(MRI) characteristics of spine and spinal cord deformity in children.Methods Thirty-three children with spine and spinal cord deformity underwent MRI examination from Jan.2010 to Dec.2012.The technique was to obtain spin echo(SE) sagittal T1WI and fast spin echo(FSE) T2WI as well as axial FSE T2WI.Coronal FSE T2WI was added when necessary.Results There were 17 cases with spine deformity,accounting for 51.5% of all patients,12 cases with both spine and spinal cord deformity,accounting for 36.4% of all patients,4 cases with sacral canal cyst,accounting for 12.1% of all patients.There were 4 cases of scoliosis,22 cases of scoliosis accompanying vertebral or spinal cord deformity,2 cases of atlas assimilation and basilar impression,1 case of bifid spine of the sacrum.Conclusions There are a great variety of paediatric spine and spinal cord deformities,and they always coexist.MRI can supply accurate diagnosis and useful information for clinic.

7.
Chinese Journal of Pediatrics ; (12): 293-296, 2010.
Article in Chinese | WPRIM | ID: wpr-245413

ABSTRACT

<p><b>OBJECTIVE</b>To explore mutation of Cited2 gene coding strand in Chinese patients with congenital heart disease (CHD).</p><p><b>METHODS</b>DNA was extracted from the blood samples of 120 nonhomologous and various CHD patients and 100 healthy children. The sequence of coding regions of Cited2 was amplified by PCR and compared to those in the GeneBank after sequencing to identify the mutations. The family of the samples who have Cited2 mutations were investigated as well. Clustal W software was applied for conservative analysis of the altered amino acids.</p><p><b>RESULTS</b>Three new mutations of Cited2 coding strand were found in 4 CHD patients. Two point mutations were first identified respectively in two patients, one patient with mirror image dextrocardia and tetralogy of Fallot (c.550 G > A), another with aortic stenosis (c.574 A > G). Apart from this, the same deletion (c.573-578del6) was first detected in another two patients, one with ventricular septal defect and atrial septal defect, the other with aortic stenosis and pulmonary stenosis. All the mutations resulted in the protein changes (p.Gly184Ser; p.Ser192Gly; p.Ser192fs). None of these changes were detected in the control group.</p><p><b>CONCLUSION</b>This study showed that there are 3 brand-new gene mutations as demonstrated by sequencing of Cited2 gene in Chinese CHD patients with a broad phenotype spectrum. Serine-glycine rich junction (SGJ) is considered as the mutation hot spot. Cited2 mutations may be one of the causes of the development of CHD in human.</p>


Subject(s)
Case-Control Studies , Child , Child, Preschool , Female , Heart Defects, Congenital , Genetics , Humans , Infant , Infant, Newborn , Male , Mutation , Repressor Proteins , Genetics , Trans-Activators , Genetics
8.
Article in Chinese | WPRIM | ID: wpr-332454

ABSTRACT

<p><b>OBJECTIVE</b>To develop a new platform for genotyping human papillomavirus(HPV) and to investigate its effect in clinical application.</p><p><b>METHODS</b>By combining L1 consensus PCR and multiplex hybridization using a Luminex xMAP system-based suspension array, we developed a rapid high-throughput assay,the HPV DNA suspension array (HPV-SA), capable of simultaneously typing 30 HPVs, including 18 high-risk HPV genotypes and 12 low-risk HPV genotypes. 810 clinical specimens were used to investigate the effect of HPV-SA. Veracity of the genotyping result was verified by E7 type-specific PCR-DNA sequencing.</p><p><b>RESULTS</b>Among the 810 clinical specimens, 243 were found to be HPV positive,including high-risk HPV subtypes 16, 18, 26, 31, 33, 39, 45, 51, 53, 56, 58, 59, 66, 68, 73 and 82,and low-risk HPV6, 11, 34, 54, 61, 67, 70 and 84. The sensitivity tested by standard samples was up to 10 copies of HPV DNA.</p><p><b>CONCLUSION</b>The HPV-SA developed here showed high sensitivity and specificity, suitable to be applied in clinical practice for HPV diagnosis and investigation on the prevalence of HPV sub-types.</p>


Subject(s)
Adult , Aged , Clinical Laboratory Techniques , Female , Genotype , Humans , Middle Aged , Nucleic Acid Hybridization , Methods , Oligonucleotide Array Sequence Analysis , Methods , Papillomaviridae , Classification , Genetics , Papillomavirus Vaccines , Classification , Genetics , Polymerase Chain Reaction , Methods , Sensitivity and Specificity , Sequence Analysis, DNA , Suspensions , Uterine Cervical Diseases , Virology , Young Adult
9.
National Journal of Andrology ; (12): 608-611, 2006.
Article in Chinese | WPRIM | ID: wpr-343562

ABSTRACT

<p><b>OBJECTIVE</b>To evaluate the role of flow cytometry in semen assessment.</p><p><b>METHODS</b>Semen samples from 104 infertile male patients (as the case group) and 10 fertilized donors (as the control group) were analyzed for the volume of ejaculate and sperm concentration, motility and atypical morphology by computer-assisted semen analysis (CASA), the viability, chromatin structure and mitochondrial membrane potential (MMP) of the sperm stained by SYBR-14/PI, AO and JC-1 respectively, and assessed with flow cytometry. The results were analyzed through SAS software.</p><p><b>RESULTS</b>U tests indicated that the semen from the infertile patients had not only lower concentration (U = 2.51, P = 0.0143), lower motility (U = 3.44, P = 0.001) and higher rate of atypical morphology (U = -5.88, P < 0.0001), but also lower viability (U = 4.72, P < 0.0001), MMP (U = -2.53, P = 0.0309), and chromatin integrity (alpha t: U = -3.82, P = 0.0003; SD alpha t: U = -3.98, P = 0.0001; COMP alpha t: U = -3.57, P = 0.0005). The multiple stepwise regression analysis revealed that sperm motility was positively correlated with sperm membrane integrity (t = 1.66, P = 0.1016), sperm MMP (t = 3.33, P = 0.0014) and sperm acrosome integrity (t = 3.24, P = 0.0019), while sperm MMP was negatively correlated with the rates of sperm neck and tail defects (t = -3.44, P = 0.001).</p><p><b>CONCLUSION</b>Flow cytometry plays a significant role in the evaluation of the quality of human sperm, and can be adopted as a useful tool in the diagnosis and treatment of male infertility.</p>


Subject(s)
Adult , Case-Control Studies , Cell Survival , Physiology , Chromatin , Pathology , Flow Cytometry , Humans , Infertility, Male , Male , Membrane Potentials , Middle Aged , Sperm Motility , Spermatozoa , Congenital Abnormalities , Pathology , Physiology
10.
Article in Chinese | WPRIM | ID: wpr-291832

ABSTRACT

<p><b>OBJECTIVE</b>To monitor the level of phthalates in human biological samples.</p><p><b>METHODS</b>The concentrations of three commonly-used phthalate (di-ethyl phthalate, DEP; di-n-butyl phthalate, DBP; di-2-ethylhexyl phthalate, DEHP) in the human biological samples were measured by using reversed-phase HPLC. The blood serum samples were collected from 52 women and 8 men, semen specimens from 36 men, and fat samples from 6 women and 5 men. All these people were randomly selected, from 23 to 50 years of age and residing in Shanghai. We also measured hormone levels of serum and conventional indices of semen specimens.</p><p><b>RESULTS</b>The three phthalates were detected in most of the biological samples, with median levels of 5.71 mg/L (ND-37.91 mg/L) in blood serum, 0.30 mg/L (0.08 -1.32 mg/L) in semen specimens, and 0.72 mg/kg (ND-2.19 mg/kg) in fat samples. The spearman correlation coefficients between concentrations of phthalates and levels of hormone in serum were 0.442 for DBP and E(2), and -0.486 for DEP and testosterone. There was a positive association between liquefied time of semen and semen concentrations of phthalates. The correlation coefficients were 0.456 for DEP, 0.475 for DBP, and 0.457 for DEHP, respectively. There was no significant difference between semen concentrations of phthalates and sperm density.</p><p><b>CONCLUSION</b>These results suggest that people residing in Shanghai area are exposed to phthalates (particularly to DBP and DEHP) though the level is still relatively low.</p>


Subject(s)
Adipose Tissue , Metabolism , Adult , Aged , China , DNA-Binding Proteins , Blood , Diethylhexyl Phthalate , Blood , Environmental Exposure , Enzyme-Linked Immunosorbent Assay , Female , Hormones , Blood , Humans , Male , Middle Aged , Phthalic Acids , Blood , Pyrrolidines , Blood , Semen , Metabolism , Transcription Factors , Blood
11.
Article in Chinese | WPRIM | ID: wpr-686747

ABSTRACT

Objective To compare and evaluate the effect of PBL in clinical teaching of Pediatrics.Methods Among students of Grade 2002 in our university,two types of PBL,pre-learning and case-discussion,were used in their clinical learning of Pediatrics. And then,their effects were evaluated and compared with those of traditional learning method.Results More than 60% of the students agreed with PBL methods,and they considered PBL favorable to practice scientific logical thinking of clinical affairs,to increase their capabilities of learning,oral expression,communication and cooperation.The teachers agreed with PBL methods too for the better learning effect resulting from PBL.Conclusion PBL fits the needs of medical learning reformation.To train new type of doctors in century 21st,it is necessary to use kinds of new learning methods,including PBL methods and standardized patient (SP)in clinical teaching.

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