Objective To explore the value and applicability of the activity of glucose-6 phosphate dehydrogenase(G-6PD) for the auxiliary diagnosis of thalassemia.Methods Nine hundred and forty samples verified by the agar gel hemoglobin electrophoresis and(or) gene diagnosis,blood count measurement,serum ferritin and G-6PD activity test were divided into 3 groups [820 cases of thalassemia in group A;40 cases of iron deficiency anemia(IDA) in group B;80 cases normal control group in group C] and the G-6PD activities of them were analyzed statistically.Results The activity of G-6PD of those samples were(35.23?7.11),(34.95?10.72),(26.64?10.85),(23.86?7.68),(19.89?5.99),(18.65?6.67),(16.75?5.49) NBTU respectively in HbH disease,?-thalassemia major,?-thalassemia intermedia,IDA,?-thalassemia minor,? combine with ?-thalassemia,?-thalassemia trait,there were significant differences compared with normal control group(Pa0.05).Conclusions G-6PD activity increase in both thalassemia and IDA group,it can be used in auxiliary diagnosis of thalassemia but had its serviceable range.It′s suitable for the auxiliary diagnosis of HbH disease and ?-thalassemia major,but not for the discrimination of gene category in thalassemia.

Animals ; Endoplasmic Reticulum ; metabolism ; Ischemia ; metabolism ; Male ; Molecular Chaperones ; metabolism ; Pressure Ulcer ; metabolism ; Rats ; Rats, Sprague-Dawley ; Subcutaneous Tissue ; blood supply

OBJECTIVETo study improvement of detection of paternally herited fetal mutant genes for β-globin in maternal plasma by PNA clamp to seek a noninvasive prenatal diagnostic method for β-thalassemia.

METHODSA total of 38 maternal blood samples were collected at 7 to 20 weeks of gestation, samples in which the father carried CD41-42 mutation and mother carried normal gene or the other point mutation for β-thalassemia were examined. The results of fetal DNA in amniotic fluid, cord blood or peripheral blood of newborns were used as the gold standard for comparison. In the study group, the total cell-free DNA was extracted from maternal plasma using QIAamp DNA Blood Mini Kit. After extraction, the total cell-free DNA was separated by agarose gel (1%) electrophoresis, and the cell-free DNA with a size of 100-300 bp was retrieved from the gel slice. Then, the retrieved DNA-free cell underwent PCR amplified with a PNA clamp. The genotype was confirmed by the conventional method (reverse dot blot hybridization), and the results were compared to gold standard. Simultaneously, two control groups with different PCR procedures were set up. The PCR procedure of control group A was amplified with the extracted total cell-free DNA and PNA clamp, and the PCR procedure of control group B was amplified with the retrieved size-fractionated DNA-free cell without PNA clamp.

RESULTSPlasma samples from 38 pregnant women were detected using PCR products for hybridization, the results were compared with the gold standard. Regarding the 21 samples confirmed by gold standard with fetal genotype 41-42M/N, 19, 8, 12 cases were detected as fetal genotype 41-42M in study group, control group A and control group B respectively, the sensitivity was 90.5% (19/21), 38.1% (8/21) and 57.1% (12/21) respectively;Concerning the 17 samples confirmed by gold standard with fetal normal genotype, the amount of false positive cases were 1, 2 and 1 respectively. The respective specificity was 94.1% (16/17), 94.1% (16/17) and 88.2% (15/17) respectively. The respective accuracies were 92.1% (35/38), 63.2% (24/38) and 71.1% (27/38) respectively. The difference in sensitivity and specificity was pairwise compared by means of McNemar's test. There was significant difference between new study group and control group A or control group B (all P﹤0.05).

CONCLUSIONThe method of detection of paternally inherited fetal mutation genes for β-thalassemia using small size of fetal DNA-free cell in maternal plasma with PNA clamp had several advantages of reliable sensitivity, specificity and accuracy, indicating its potential of clinical practicality.

Adolescent ; Adult ; DNA ; blood ; Female ; Humans ; Inheritance Patterns ; Mutant Proteins ; genetics ; Mutation ; Peptide Nucleic Acids ; Pregnancy ; Prenatal Diagnosis ; Young Adult ; beta-Globins ; genetics ; beta-Thalassemia ; diagnosis ; genetics

Objective To explore the change of dietary intake and nutritional status before and after hematopoietic stem cell transplantation (HSCT) in pediatric patients to assess the importance of nutritional interventions.Methods In this observational cohort study,65 children undergoing HSCT between January 2012 and November 2012 in the Department of Hematology and Oncology,Shanghai Children's Medical Center were enrolled.The data collected before preconditioning were considered as the baseline data.We also collected data twice a week between preconditioning and 30 days after HSCT,and once a week from 30 days to 100 days after HSCT.Dietary analysis and urea nitrogen analysis were conducted in parallel.Results The baseline level of energy intake was (5 844.9 ±2 490.4) kJ/d,protein intake was (56.4 ±28.6) g/d,fat intake was (49.7 ±38.9) g/d,and carbohydrate intake was (190.9 ± 91.1) g/d.With the hematopoietic reconstruction,the oral nutrients intake significantly decreased compared with the baseline levels (all P =0.000).During the recovery period after HSCT,the energy intake showed no significant difference when compared with the baseline level in the 6th postoperative week,protein in the 13th week,carbohydrate in the 4th week,and fat in the 6th week.The urine nitrogen was (3.9 ± 2.4) g/d before HSCT,which increased to (16.7 ± 11.0) g/d after preconditioning (P=0.000).In the 1st postoperative week,the weight (P =0.000),triceps skin fold thickness (P =0.003),mid-upper arm circumference (P =0.000),serum albumin (P =0.000) and prealbumin (P =0.000) of the patients all significantly decreased compared with the baseline levels.In the 9th postoperative week,the fat-free body weight percentage (P =0.010),muscle percentage (P =0.001) and protein percentage (P =0.000) were significantly lower than the baseline levels,while the body fat percentage was higher than the baseline level (P =0.002).Conclusions Children undergoing HSCT exhibit a marked reduction in nutrient intakes at the early stage of HSCT,which may gradually return normal during the recovery period.This process may be slow,especially for the protein,and therefore may affect the serum protein level in these pediatric patients.Thus,more careful nutrition guidance is necessary during HSCT for pediatric patients,emphasizing oral nutrients intakes,and high protein dietary or formula may be helpful.

AIM: To compare the effectiveness of botulinus toxin of type A and complete resection of the periorbital muscle on idiopathic blepharospasm.
● METHODS: Patients with idiopathic blepharospasm and having undergone either of two procedures from Dec. 2010 to Jun. 2015 were selected ( 60 patients ) . Among them, group A (30 patients, 60 eyes) underwent botulinus toxin of type A, group B (30 patients, 60 eyes) underwent complete resection of the periorbital muscle.
●RESULTS: ln group A, the patients with complete response, obvious response, partial response, and no response were 36(60. 0%), 20(33. 3%), 2(3. 3%) and 2 (3. 3%) cases respectively. ln group B, the patients with complete response, obvious response, partial response, and no response were 16(26. 7%), 24(40. 0), 12(20. 0%) and 8 ( 13. 3%) cases respectively. The difference was statistically significant ( Z = - 2. 968, P = 0. 003 ). The relapse rate of group A and group B were 93. 3% and 20. 0% after 6mo, the difference was statistically significant (χ2=32. 851, P<0. 001).
●CONCLUSION: The botulinus toxin injection of type A is effective for idiopathic blepharospasm. But recurrence rate is high after 6mo. Complete resection of the periorbital muscle have long-term efficacy for idiopathic blepharospasm. It′s a supplementary therapy after idiopathic blepharospasm recurrence.

Objective To explore the effect of aminolevulinic acid-based photodynamic therapy(ALA-PDT) on alloreactived peripheral blood mononuclear cells(PBMCs).Methods Human PBMCs from different healthy donor were collected and mixed in the one-way mixed lymphocyte culture(MLC) for 5 days. The cells were harvested and aminolevulinic acid(ALA) were added into ALA group and ALA+Light group with ultimate concentrations of 0.5 mmol/L,1.0 mmol/L,1.5 mmol/L,2.0 mmol/L and 2.5 mmol/L.After cultured for 2 hours, 4 hours and 6 hours respectively in 37 ℃ 5% carbon dioxide incubator,Light group and ALA+Light group were irradiated by light of 410 nm wavelength for 1 hour.The MLC cells were treated with the former stimulator cells for 48 hours.The survival of stimulator cells were detected using MTT colorimetric assay and the kill rates of treated cells were calculated.Results The kill rate of ALA+Light group on stimulators was apparently lower than those of Light group, ALA group and control group, (33.0?26.5)% vs (87.1?2.2)%,(89.2?2.5)%,(90.3?1.9)%(All P

Objective To observe the effect of Guanxinning Tablet (GXNT) on myocardial infarction and cardiac autonomic nervous function in rats with myocardial ischemia reperfusion injury (MI/RI).Methods Seventy SD rats were divided into 7 groups randomly (n=10);the sham group, the MI/RI group, 75 mg/kg, 150 mg/kg, 300 mg/kg and 600 mg/kg GXNT groups and 300 mg/kg Compound Danshen Tablets (DST) group.All rats were administered orally for 7 days, and then the MI/RI model was made by ligating the left anterior descending branch of coronary artery in rats.The changes of electrocardiogram were recorded and the electrocardiogram of J points and heart rate variability (HRV) parameters were analyzed.At the end of reperfusion, the myocardial infarct size was measured by using Evans blue and tetrazolium chloride (TTC) double staining, and pathological changes of myocardium were observed by HE staining.The changes of serum lactate dehydrogenase (LDH), creatine kinase (CK), malondialdehyde (MDA) and nitric oxide (NO) levels were also detected.Results Compared with MI/RI group, GXNT and DST groups were significantly reduced myocardial infarct size and inhibited the rising of serum LDH and CK activities (P < 0.05, P < 0.01), and also reduced the total or average value of J point during reperfusion (P < 0.05, P < 0.01).Meanwhile, GXNT and DST groups were markedly increased HRV and serum NO level as well as decreased serum MDA content (P < 0.05, P < 0.01), and improved myocardial tissue pathology.Conclusions GXNT can reduce the myocardial infarction in rats with MI/RI, and also improve the cardiac autonomic nervous function.

Purpose To evaluate the correlations between high resolution CT (HRCT) findings and IASLC/ATS/ERS pathological classification of ground glass nodule (GGN). Materials and Methods 121 patients with confirmed GGN were selected, and divided into benign group (22 cases), PIL group (21 cases), microinvasive carcinoma group (26 cases) and invasive carcinoma group (52 cases), then the imaging, pathology and prognosis data of patients with pulmonary GGN were reviewed, and the differences among GGN of different pathological types were analyzed.Results Maximum diameter, margin, vacuole sign, solid component, shape and blood vessels through of GGN were significantly different among the four groups (χ2=9.945-31.068,P<0.05). Maximum diameter and margin were significantly different between invasive adenocarcinoma and other groups (P<0.008); vacuole sign of the benign group was significantly different with other groups (P<0.008); the existence of solid component and shape were significantly different between invasive adenocarcinoma and minimally invasive adenocarcinoma (P<0.008); there was significant difference of blood vessels through between invasive adenocarcinoma and benign lesions (P<0.008). Among the 121 lesions, no metastasis except one invasive adenocarcinoma case complicated with distant metastasis.Conclusion Maximum diameter of GGN greater than 16.35 mm, with spiculation or lobulation represent invasive adenocarcinoma; vacuole sign within the GGN represent malignancy; with solid component and irregular shape can be used to identify invasive adenocarcinoma from minimally invasive adenocarcinoma; while blood vessels through can be used to identify invasive adenocarcinoma from benign lesions; the prognosis of GGNs is well with only 0.83% probability of distant metastasis.

OBJECTIVENYD-SP5 is a newly cloned gene highly expressed in human testes, which consists of 3 598 nucleotides including a 1 027-amino acid open reading frame. It is a human-mouse homologous gene. The domain analysis indicated that the NYD-SP5 protein is a transmembrane protein. This study aimed to design and establish recombinant plasmids of small hairpin interfering RNA (shRNA) against NYD-SP5, and to pave the way for the analysis of the function of NYD-SP5 in the testis using the transgenic mouse model.

METHODSFour sequences of oligonucleotides with the small hairpin structure were designed based on the NYD-SP5 mRNA sequence. Recombinant plasmids were constructed by cloning these oligonucleotides into pGPU6/GFP/Neo vectors. Interfering plasmids against GAPDH were established as positive controls and those targeting non-specific genes used as negative controls. The positive constructs were verified by enzyme digestion and sequencing.

RESULTSPlasmid screening and sequencing showed the sequences of the recombinant plasmids to be the same as the shRNA transcribed sequences, which indicated the successful establishment of the recombinant vectors.

CONCLUSIONThe shRNA expression vector targeting NYD-SP5 could be established successfully.

Animals ; Base Sequence ; Genetic Vectors ; Humans ; Male ; Mice ; Mice, Transgenic ; Molecular Sequence Data ; Plasmids ; Proteins ; genetics ; RNA, Small Interfering ; genetics

To explore the antagonistic effect of gingerols against the inflammation induced by lectin from Pinellia ternata. In this study, ELISA method was used to determine the effect of different extracts from gingerols on the release of inflammatory factor TNF-α from macrophages induced by lectin from P. ternata. The fluorescence probe was used to determine the effect of gingerols on the changes in ROS of macrophages induced by lectin from P. ternata. The western-blot method was applied to study the effect of gingerols on the increase in expression of cell receptor interacting protein RIP3 in macrophages induced by lectin from P. ternata. The scanning electron microscope (SEM) was used to study the effect of gingerols on morphological changes in macrophages induced by lectin from P. ternata. According to the results, gingerols can significantly inhibit the release of inflammatory factor from macrophages induced by lectin from P. ternata, ROS overproduction and increase in RIP3 expression. SEM results showed that gingerols can inhibit the cytomorphosis and necrocytosis induced by lectin from P. ternata. Fresh ginger's detoxication may be related to gingerols' effects in inhibiing release of inflammatory factor, ROS overproduction and increase in RIP3 expression caused by macrophages induced by lectin from P. ternata, which are mainly inflammatory development.
Animals ; Catechols ; pharmacology ; Cells, Cultured ; Drug Antagonism ; Fatty Alcohols ; pharmacology ; Ginger ; chemistry ; Lectins ; toxicity ; Macrophages ; drug effects ; metabolism ; Male ; Mice ; Mice, Inbred ICR ; Pinellia ; chemistry ; toxicity ; Reactive Oxygen Species ; metabolism ; Receptor-Interacting Protein Serine-Threonine Kinases ; genetics ; metabolism ; Tumor Necrosis Factor-alpha ; genetics ; metabolism