Objective To study the diagnostic significance and clinical value of genetic analysis in children with neonatal diabetes. Methods Gene mutation analysis was performed in four patients from Zhengzhou children ' s hospital with diagnosis of with neonatal diabetes. Therapeutic effect of glibenclamide in patients with or without gene mutation was compared. Results KCNJ11 gene mutation was found in two patients with neonatal diabetes. Glibenclamide was found only effective for blood glucose control in patients with KCNJ11 mutation. Therefore, Insulin remains the best therapeutic choice in patients without the genetic mutation. Conclusions Genetic mutation status may be useful in choosing treatment options of neonatal diabetic patients, therefore, should be performed in all children with neonatal diabetes.

The aim of this study was to detect the localization and level of tyrosine phosphorylated proteins during in vitro capacitation of guinea pig sperm. Sperm from mature guinea pigs were incubated in modified TALP under 5% CO_2 in air at 37 ℃. The capacitation effect was assessed by chlortetracycline (CTC) staining. Western blotting and indirect immunofluorescence were used to analyze the level and localization of tyrosine phosphorylation. The results showed that guinea pig sperm underwent a time-dependent increase in protein tyrosine phosphorylation during the in vitro capacitation and the percentage of protein tyrosine phosphorylated sperm increased from 36% to 92% from the beginning of incubation to 7h incubation. Also, there was a shift in the site of phosphotyrosine-specific fluorescence from the head of sperm to both the head and the flagellum of sperm. Moreover, there were three proteins phosphorylated in this experiment. After 0 to 0.5h incubation, the protein of 40kDa was detected by anti-phosphotyrosine monoclonal antibody, and the intensity of this protein increased in the following incubation. Then, after 1h incubation, another protein of 80kDa was found and the level of this protein reached the highest point at 3h. Also, in 3h incubation, a protein of 45kDa was detected and the intensity of this protein increased in the following incubation.

Aim To investigate the antioxidant capacity of crocetin and crocin in an in vivo system.Methods Column chromatography was applied to the seperation of crocetin and crocin-1 from gardenia.Crocetin(6.25,12.5 and 25.0 mg·kg~(-1)·d~(-1)) and crocin (18.7,37.5 and 75.0 mg·kg~(-1)·d~(-1)) were orally administered to kunming mice.Then,superoxide dismutase(SOD),glutathione peroxidase(GSH-Px),total antioxidant capacity(TAOC)and malondialdehyde(MDA)in mice were determined for the comparison of antioxidant activity of crocetin and crocin-1.Results Oral administration of crocetin and crocin for six weeks could enhance SOD of liver and kidney,GSH-Px of liver and TAOC of heart and kidney.In addition,it could decrease MDA of serum in mice.Conclusions The comparison of results suggests the evidence supporting the comparable antioxidant activity of crocetin and crocin.The results of the research also indicate that liver and kidney are two organs targeted for protection concerning endogenous antioxidant among various tissues.

OBJECTIVETo assess the efficacy of tendons of minimally invasive therapy (TMIT) combined drug therapy by comparing it with treatment by drug therapy alone on patients with knee osteoarthritis (KOA).

METHODSTotally 60 KOA patients were assigned to the treatment group and the control group according to random digit table, 30 in each group. Patients in the control group took Hydrochloric Acid Glucosamine Capsule and Celecoxib Capsule. Patients in the treatment group additionally received TMIT. The treatment course for all was 4 weeks. Scores for visual analogue scale (VAS) and the Western Ontario and McMaster Universities (WOMAC) Osteoarthritis Index were observed and recorded at week 1 and 4 after treatment by acupotomology mirror.

RESULTSCompared with before treatment, improvement was shown in VAS score, pain and stiffness degrees, activities and functions, and WOMAC scores at week 1 and 4 after treatment in all patients with statistical difference (P < 0.05). Besides, better effect was shown in the treatment group (P < 0.05).

CONCLUSIONSTMIT combined drug therapy could relieve KOA patients' pain, stiffness and joint activities, elevate the overall efficacy. TMIT was easily operated with less injury.

Celecoxib ; Drug Therapy, Combination ; methods ; Humans ; Knee Joint ; Osteoarthritis, Knee ; drug therapy ; Pain ; Pain Measurement ; Tendons ; Treatment Outcome

OBJECTIVETo observe the the expression of endoplasmic reticulum stress (ERS) related factors in deep tissue injury (DTI) at pressure ulcer rat and to investigate the ERS mechanism of DTI in muscle tissue and protective effect of 4-phenylbutyric acid (4-PBA) in local tissue.

METHODSFifty male SD rats were randomly devided into control group, model group, experimental group NS group and PBA group, the experimental groups were divided into 4 d, 7 d, 14 d and 21 d group according to the observation time (n = 5). Rats in the PBA group were administrated with gastric perfusion of 4-PBA after the modeling; the NS group was given normal saline of the same quantity. Using HE staining to observe morphologic character. The expression of glucose regulated protein 78 (GRP78), CHOP, Caspase 12 were detected by immunohistochernical staining. Cell apoptosis was detected by TUNEL assay.

RESULTSHE staining results showed that each group demonstrated compression injury compared with control group: cellular swelling, ompaction of nuclear, and apoptosis in muscle tissue. The new muscle fiber in 4-PBA group fused faster than those in NS group. The number of TUNEL positive cells peaked at 4 day after compression, then got decreased on day 7 in muscle tissue, apoptosis positive cells were diminished after 4-PBA treatment. The immunohistochemical staining results showed that the expression of protein GRP78, CHOP, Caspase 12 peakd 4 d after modeling and decreased gradually. The GRP78, CHOP, Caspase 12 protein expression were significantly higher than those of PBA group at all time points (P < 0.05).

CONCLUSIONCell apoptosis induced by endoplasmic reticulum stress took part in deep tissue injury resulting of pressure ulcer, which mechanism might be related to reducing apoptosis mediated by CHOP, Caspase 12.

Animals ; Apoptosis ; Caspase 12 ; metabolism ; Endoplasmic Reticulum Stress ; Heat-Shock Proteins ; metabolism ; Male ; Muscle, Skeletal ; pathology ; Phenylbutyrates ; pharmacology ; Pressure Ulcer ; physiopathology ; Proteomics ; Rats ; Rats, Sprague-Dawley ; Transcription Factor CHOP ; metabolism

The objective of this study was to investigate the immunophenotype of T-lineage acute lymphoid leukemia (T-ALL) and to find valuable immunologic markers in T-ALL diagnosis and therapy. Four-color multiparametric flow cytometry(FCM) with CD45/SSC gating was used for immunophenotyping of 95 patients with newly diagnosed T-ALL. The results demonstrated that T-ALL occurred more frequently in males younger than 30 years of age and was usually accompanied by a high WBC count and tumor mass at diagnosis. Univariate analysis showed an influence on achievement of CR1 for age (< 30 years) but not for WBC count and tumor mass. According to WHO (2008) classification of tumors of haematopoietic and lymphoid tissues, 87 patients with confirmed subtype included 27 cases of Pro-T-ALL (31.0%), 31 cases of Pre-T-ALL (35.6%), 23 cases of cortical-T-ALL (26.4%), 6 cases of medullary-T-ALL (6.9%). CD34 expression in Pro-T-ALL was significantly higher than that of Pre-T-ALL (p = 0.001). After the first chemotherapy, the complete remission rate in Pro-T-ALL was statistically lower than that of Pre-T-ALL. Besides, the complete remission rate of immature T-ALL (including Pro-T-ALL and Pre-T-ALL) was also significantly lower than that in mature T-ALL (including cortical-T-ALL and medullary-T-ALL). Myeloid antigen (CD13, CD33) expression was associated with T-ALL subtype and treatment effect. While 66.7% of CD13(+) patients belonged to Pre-T-ALL, most (60.0%) of CD33(+) patients were classified into Pro-T-ALL; CD13 expression had no effect on CR1 rate whereas CD33(+) patients had worse treatment effect compared with CD33(-) groups (p = 0.001). Notably, the expression of CD117 reached up to 26.7% and the positive cases were primarily distributed in pro-T-TAll and pre-T-ALL. It is found that CD117 expression in CD34(-) group was homogeneous and CD117 expression level was less than 10% in 73.2% patients, but CD117 expression level in CD34(+) group was not homogenous, in which group the CD117 expression levels < 10%, 10% - 20% and > 20% were 44.2%, 17.3% and 38.5% respectively. As compared with CD34(-) group, the proportion of patients with CD117 expression levels < 10%, > 20% in CD34(+) group was higher, and there was significant difference between these 2 group. It is concluded that immunophenotype has great value in T-ALL diagnosis, classification as well as treatment. Flow cytometry provides access to find valuable immunologic markers for T-ALL biological research.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; CD13 Antigens ; metabolism ; Child ; Child, Preschool ; Female ; Humans ; Immunophenotyping ; Infant ; Male ; Middle Aged ; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma ; classification ; immunology ; therapy ; Proto-Oncogene Proteins c-kit ; metabolism ; Sialic Acid Binding Ig-like Lectin 3 ; metabolism ; Young Adult

OBJECTIVETo explore the clinical effects of total vertebral column resection combined with anterior mesh cage support in treating severe congenital kyphoscoliosis.

METHODSFrom April 2008 to April 2012,21 patients with severe congenital kyphoscoliosis were treated with total vertebral column resection and internal fixation through posterior approach combined with anterior mesh cage support. There were 8 males and 13 females with an average age of 19.4 years old (ranged from 10 to 35). And 6 cases were thoracic segments deformity,13 cases were thoracolumbar segments and 2 cases were lumbar segments, of them, 2 cases were accompanied with Chairs deformity, 6 cases with diastematomyelia, 4 cases with syringomyelia,and 1 case with neurofibromatosis. According to the Frankel grade system, 3 cases were grade C, 5 cases grade D and 13 cases grade E. Blood loss, operative time, and perioperative complications were recorded. Coronal and sagittal Cobb angle, apical vertebral offset distance, sagittal offset, the relative height of shoulders, razor back deformities were measured and analyzed before and after operation.

RESULTSThe average operative time was 5.2 h (3.5 to 6.5 h) and blood loss was 2,500 ml (1,400 to 4,900 ml). The 2nd day after operation, apical vertebral offset distance, sagittal offset, the relative height of shoulders, razor back deformities had obviously improved than preoperative (P < 0.05). There was no significant difference in above items between postoperative on the 2nd day and final follow-up (P > 0.05). The corrective rate of kyphosis and scoliosis were (60.97 +/- 6.30)% and (62.24 +/- 5.82)%, respectively. On the first day after surgery,2 cases of Frankel grade E aggravated to grade D, and obtained recovery at 2 week after conservative treatment. And 1 case palinesthesia later,grade D aggravated to grade C and obtained recovery after revision surgery in time. One case complicated with permanent blindness of left eye, 1 case occurred injury of pleura and 2 cases had cerebrospinal fluid leak during operation. All patients were followed up from 9 to 31 months with an av- erage of 18.6 months. At final follow-up,all patients obtained bone union, Frankel grade D in 4 cases and grade E in 17 cases, no correction loss and internal fixation loosening was found.

CONCLUSIONTotal vertebral column resection combined with anterior mesh cage support can effectively correct kyphosis and scoliosis in severe congenital kyphoscoliosis and can avoid injury of spine cord by spinal crispation, but intraoperative position and neurologic complications should still be considered.

Adolescent ; Adult ; Child ; Female ; Humans ; Kyphosis ; complications ; congenital ; diagnostic imaging ; surgery ; Male ; Retrospective Studies ; Scoliosis ; complications ; congenital ; diagnostic imaging ; surgery ; Spine ; surgery ; Tomography, X-Ray Computed ; Young Adult

The aim of this study was to investigate apoptotic effect of 2-methoxyestradiol (2-ME2) on K562 cells and its mechanism. K562 cells were treated with different concentrations of 2-ME2. MTT assay was used to examine the effect inducing growth inhibition. DNA fragmentation assay and Annexin V-FITC/PI staining were used to detect the effect of apoptosis. The change of mitochondrial transmembrane potential was analyzed by flow cytometry. The expressions of related gene mRNA and/or proteins were detected by RT-PCR and Western blot respectively. The results indicated that the 2-ME2 inhibited proliferation of K562 cells in a time- and dose-dependent manners and the concentration of 50% growth inhibition (IC(50)) was 2 micromol/L at 48 hours. 2-ME2 induced DNA ladder and significantly increased apoptosis in K562 cells when exposed to 2 micromol/L of 2-ME2 for 24, 48 and 72 hours, the result of Annexin-V/PI staining showed that rates of the apoptotic cells were 13.78%, 22.32% and 29.43% respectively, which was remarkably higher than that of control (1.78%) (p < 0.05). The FCM analysis showed that the mitochondrial transmembrane potential in K562 cells lowered after exposed to 1, 2 and 4 micromol/L of 2-ME2 for 24 hours. 2-ME2 down-regulated the expression of bcr/abl and bcl-2, up-regulated the expression of bax mRNA, and down-regulated protein expressions of bcl-2, procaspase-3, procaspase-9, PARP (116 kD) and p-Akt, and up-regulated expression of cytoplasmic Cyto-C and PARP 85 kD apoptosis-related cleavage fragment protein, but had no effect on total Akt protein in K562 cells after treated with 2 micromol/L of 2-ME2 for 24, 48 and 72 hours. It is concluded that the 2-ME2 can induce the apoptosis and inhibit the proliferation of K562 cells by increasing the ratio of bax/bcl-2, reducing the mitochondrial transmembrane potential, releasing cytochrome C to cytoplasm, initiating the mitochondrial apoptosis pathway and leading in turn to caspase-3 activation. These findings suggest that interfere PI3K/Akt signal pathway via down-regulating the expression of bcr/abl mRNA is implicated in the effect of 2-ME2 on K562 cells.
Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Cytochromes c ; metabolism ; Down-Regulation ; Estradiol ; analogs & derivatives ; pharmacology ; Humans ; K562 Cells ; Proto-Oncogene Proteins c-akt ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; RNA, Messenger ; genetics

OBJECTIVETo study the status of growth and development and the relationship between growth disorders and iron overload in children with beta-thalassemia major.

METHODSFifty children with beta-thalassemia major and who received blood transfusion therapy regularly (age: 9 months-17 years) were enrolled. They were subjected to a thorough history taking, clinical examinations, and laboratory examinations, including complete blood count, alanine transferasa (ALT) and serum ferritin. The physical growth parameters, such as height and weight, were compared with the reference values of Chinese children.

RESULTSTwenty-four patients (48%) were of short stature with height under the 3th percentile. Among them, 15 cases presented with their height and weight both under the 3th percentile. Spontaneous sex development was seen in 7 cases out of 21 over 10-year-old patients. No sex development was found in 4 out of 8 patients who were over 14 years old. The patients with a height under the 10th percentile (n=31) had higher serum ferritin levels (8239.2+/-5865.5 mg/L vs 5028.1+/-3885.7 mg/L; P<0.05) and lower hemoglobin levels (68.2+/-12.3 g/L vs 79.7+/-14.5 g/L; P<0.05) as well as hepatomegaly when compared with those patients with a height over the 10th percentile (n=19). Serum ferritin levels in 20 patients with a weight under the 10th percentile were significantly higher than those in 30 patients with a height over the 10th percentile (9165.5+/-6042.5 mg/L vs 5567.3+/-4447.3 mg/L; P<0.05).

CONCLUSIONSShort stature, low weight and sex development delay are common in children with beta-thalassemia major. This may be related to iron overload.

Adolescent ; Blood Transfusion ; Child, Preschool ; Female ; Growth Disorders ; etiology ; Humans ; Infant ; Iron Overload ; complications ; Male ; Sexual Maturation ; Young Adult ; beta-Thalassemia ; complications ; therapy