ObjectiveTo explore the prevalence and spectrum of β-thalassemia mutations in Fujian province,and to provide a reference for prenatal diagnosis and genetic counseling in this population.Methods Two thousand three hundred and one blood samples were randomly selected from 9 different areas of Fujian province from May 2008 to December 2010.PCR and reverse dot blot hybridization (RDB) were adopted for detection of the 17 common types of mutation,and the frequency of each genotype of β-thalassemia mutations was calculated.The β-globin gene of unknown positive samples were analyzed directly with DNA sequencing.Results Three hundred and fifty-nine cases were detected with β-thalassemia mutations of the 2301 copy blood samples submitted,and the detection rate was 15.60％ (359/2301).Of the mutated genes,12 different mutations were identified,namely IVS-2-654(C→T),CD41-42(-TCTT),CD17(A→T),-28(A→G),CD27-28(+C),CD26(G→A),CD71-72(+A),IVS-1-1(G→T),CD43(G→T),-29(A→G),initiation codon ATG→AGG and CD36(-C).Mutation frequencies were 46.54％ (175/376),33.24％ (125/376),9.31％ (35/376),5.05％ (19/376),2.13％(8/376),1.33％(5/376),0.80％(3/376),0.27％(1/376),0.27％(1/376),0.27％(1/376),0.53％(2/376),and 0.27％(1/376),respectively.The most common mutations were IVS-2-654 (C→T) and CD41-42 (-TCTT),which accounted for 79.78％(300/376) of total genetic mutations.In addition,a novel β-globin gene mutation CD36 (-C) allele was detected for the first time,the deletion of a nucleotide C at code 36 within exon 2 lead to a frameshift mutation that could result in a premature termination at code 60.Conclusions β-thalassemia mutations in Fujian province are complex with significant genetic heterogeneity.We present for the first time the detection of a new β-thalassemia mutation in the population:CD36(-C),which provides valuable information for genetic counseling and prenatal diagnosis in Fujian province.

Objective To observe the effect of different thyroid hormone level on the expression of synaptotagmin Ⅰ(Syt Ⅰ) in adult rat hippocampus. Methods All 28 adult male SD rats were assigned randomly into hypothyroid, hyperthyroid and control group, hypothyroid group was established by daily intraperitoneal injections with propylthiou raci(PTU, 10.0 mg/kg body weight) for 6 weeks and hyperthyroid group with L-Thyroxine (L-T4, 0.5 mg/kg body weight) for 3 weeks. Radioimmunity method was used to assay the levels of serum T3 and T4, immunohistochemical S-P technology to assay the levels of Syt Ⅰ protein in hippoeampus CA1, CA3 and dentate gyrus (DG). The layers analyzed in the different subfields include the polymorphic cell layer(the stratum oriens, SO), pyramidal cell layer(PCL), stratum radiatum (SR), lacunosum-molecular layer (SLM) in CA1 and CA3, granular cell layer(GL) and molecular layer(ML) in DG. Results The levels of serum T3 and T4[(0.34±0.12), (41.03± 11.37)nmol/L]in the hypothyroid rats were significantly lower than those in the control group[(0.65±0.15), (55.20±10.68)nmol/L, P < 0.01 or < 0.05], and the positive granule of Syt Ⅰ was significantly lower in PCL and SR of CA1 and CA3, GL of DG. The average optical value responsible for Syt Ⅰ immunoreactivity was obviously reduced in SO(0.048±0.007), PCL(0.299±0.035), SR(0.042±0.007), SLM(0.038±0.006) of CA1, PCL(0.085± 0.019), SR(0.040±0.011), SLM (0.038±0.006) of CA3, GL (0.076±0.019) of DG than normal controls (0.068± 0.014, 0.376±0.053, 0.053±0.008,0.056±0.009,0.118±0.026,0.052±0.010,0.053±0.009,0.099±0.015; P< 0.01 or < 0.05). Serum T3 and T4 levels [(1.43±0.30), (157.18±19.95)nmol/L]of hyperthyroid rats were significantly higher than those of control group(P < 0.01). The value was reduced in PCL(0.322±0.050), SR(0.039±0.006), SLM (0.042±0.006) of CA1, PCL(0.098±0.034), SR(0.046±0.013), SLM(0.046±0.010) of CA3 and GL(0.085± 0.024), ML (0.042±0.009) of DG (P < 0.05 or < 0.01). Conclusion Adult-onset of hypothyroidism and hyperthyroidism can reversibly decrease the expression of Syt Ⅰ in CA1, CA3 and DG regions of hippocampus.

OBJECTIVETo evaluate the effect of palmitic acid (PA) on oxidative stress and activation of inflammasomes in hepatocytes.

METHODSTo test the dose-dependent effect of PA on normal murine hepatocytes AML12, the cells were treated with 0, 0.15, 0.25 and 0.4 mmol/L of palmitic acid (PA). The cells were also divided into blank control group, 0.25 mmol/L PA group and 0.25 mmol/L PA+N-acetylcysteine (NAC) group to examine the effect of reactive oxygen species (ROS) on the activation of inflammasomes. After 24 h of treatment, lipid accumulation, total ROS, mitochondrial ROS, expression and localization of NOX4, and expressions of inflammasomes and IL-1β were detected in the hepatocytes.

RESULTSCompared with the control cells, PA treatment of the cells significantly increased cytoplasmic lipid accumulation, concentrations of total ROS (12 463.09±2.72 vs 6691.23±2.45, P=0.00) and mitochondrial ROS (64.98±0.94 vs 45.04±0.92, P=0.00), and the expressions of NOX4, NLRP3, ASC, caspase-1, and IL-1β (1603.52±1.32 vs 2629.33±2.57, P=0.00). The mitochondria and NOX4 were found to be co-localized in the cytoplasm. NAC obviously reduced cellular ROS level stimulated by PA (7782.15±2.87 vs 5445.6±1.17, P=0.00) and suppressed the expressions of NLRP3, ASC and caspase-1.

CONCLUSIONPA treatment can stimulate lipid accumulation in hepatocytes and induce oxidative stress through NOX4 and mitochondria pathway to activate inflammasomes and stimulate the secretion of IL-1β.

Acetylcysteine ; pharmacology ; Animals ; Carrier Proteins ; metabolism ; Caspase 1 ; metabolism ; Cells, Cultured ; Hepatocytes ; drug effects ; metabolism ; Inflammasomes ; drug effects ; metabolism ; Interleukin-1beta ; metabolism ; Mice ; Mitochondria ; drug effects ; NADPH Oxidase 4 ; NADPH Oxidases ; metabolism ; NLR Family, Pyrin Domain-Containing 3 Protein ; Oxidative Stress ; Palmitic Acid ; pharmacology ; Reactive Oxygen Species ; metabolism

Objective To evaluate the clinical effects and toxicity of escitalopram oxalate versus paroxetine in the treatment of unipolar depre-ssion.Methods 71patientswithconfirmeddiagnosisofunipolardepre-ssion ( UD ) were recruited and randomized divided in to escitalopram oxalate ( EO ) group with escitalopram oxalate 10 mg? d-1 ( max 20 mg? d-1 ) orally, and paroxetine ( PA ) group with paroxetine 20 mg? d-1 ( max 50 mg? d -1 ) orally, with 12 weeks treatment.At the time point of 2 , 4 , 8 and 12 -week the hamilton depression scale (HAMD), self-rating depression scale (SDS) and treatment emergent symptom scale ( TESS ) score were recorded and compared between the two groups. Results The HAMD and SDS score were significant decreased ( P<0.05 ) after 4 weeks treatment in the EO group , and sig-nificant decreased ( P<0.05 ) after 2 week in the PA group.The clinical efficacy was 72.97%and 70.27% according to HAMD and SDS in the EO group.The clinical efficacy was 70.59% and 67.65% according to HAMD and SDS in the PA group .No statistical difference was found between the two groups for the clinical efficacy (P>0.05).There was no significant difference between two group on incidence of adverse drug reaction ( P>0.05 ).Conclusion There was no significant difference for escitalopram oxalate and paroxetine in the treatment of unipolar depression , but paroxetine work quickly than escitalopram oxalate .

OBJECTIVETo evaluate the efficacy and safety of tourniquet in total knee arthroplasty.

METHODStudies on comparison between with and without tourniquet in total knee arthroplasty were identified from Medline, PubMed, EMASE, Cochrane Library, CBM, Highwire, CNKI, VIP, Articles Digital Periodicals.All the randomized controlled trials were included for meta-analysis with RevMan 4.2.2 software.

RESULTSNineteen studies involving 15 in foreign languages, 4 in Chinese were identified. There were 1159 cases of knee replacement patients. The results of meta-analysis indicated that there were statistical difference between two groups on intraoperative blood loss (P = 0.000), the number of deep venous thrombosis (P = 0.020), thigh pain (P = 0.000), knee hematoma (P = 0.030), wound infection (P = 0.040), skin ecchymosis area (P = 0.000), and the increasing rate of knee circumference of 3 days after the operation (P = 0.000), while there were no statistical differences with respect to the total blood loss (P = 0.100), the number of blood transfusions (P = 0.150), operation time (P = 0.120), length of hospital stay (P = 0.350), the number of pulmonary embolism (P = 0.310), and skin blisters (P = 0.170).

CONCLUSIONSThe tourniquet for total knee arthroplasty can reduce intraoperative blood loss, but can not reduce total blood loss and the number of blood transfusions transfusion, can not improve operative efficiency, can not shorten the hospitalization time and promote the knee joint functional recovery. Furthermore the tourniquet increases the probability of occurrence on deep vein thrombosis, wound infection, hematoma and ecchymosis knee, it also causes knee swelling and thigh pain. It suggests minimize to use tourniquet in total knee arthroplasty.

Arthroplasty, Replacement, Knee ; adverse effects ; methods ; Humans ; Safety ; Tourniquets ; adverse effects

OBJECTIVETo improve the diagnosis and treatment of testicular teratoma in children by analysis of clinical data.

METHODSWe retrospectively analyzed the clinical data about 64 cases of testicular teratoma treated in the Children's Hospital of Chongqing Medical University from 1995 to 2014.

RESULTSSixty-one of the cases presented painless scrotal mass with a sense of bearing down and the other 3 cases were confirmed because of empty scrotum diagnosed as cryptorchidism. The level of serum alpha fetal protein ( AFP) was obviously increased in 46 cases but normal in the other 18 preoperatively. Ultrasonography manifested abnormal inhomogeneous echo zones with calcification or necrosis. X-ray examination presented patchy or curvilinear high-density shadows in 28 cases. Forty-one of the patients underwent testis-sparing surgery (TSS) , 20 received high inguinal orchiectomy, and 3 refused surgical treatment. Pathological examination revealed 3 mature germinal layers in the 49 cases of mature teratoma and immature germinal tissue, including the original neural tube, and 11 cases of immature teratoma. The mature cases were exempted from chemotherapy, while the immature cases received the combination of cisplatin, etoposide, and bleomycin (PEB). The patients were followed up for 2 years postoperatively, which revealed no recurrence or metastasis.

CONCLUSIONMost children with testicular teratoma presented painless scrotal mass with a sense of bearing down and with abnormal serum AFP in most cases. Ultrasonography and plain radiography of the scrotum contribute to the diagnosis of the tumor. TSS is the main treatment option and intraoperative frozen-section can help the surgeons decide on the surgical mode. Postoperative chemotherapy is necessitated for immature teratoma but not for mature cases.

Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Bleomycin ; administration & dosage ; Child ; Cisplatin ; administration & dosage ; Cryptorchidism ; diagnosis ; Etoposide ; administration & dosage ; Gonadal Dysgenesis, 46,XY ; diagnosis ; Humans ; Male ; Orchiectomy ; methods ; Retrospective Studies ; Scrotum ; Teratoma ; blood ; diagnosis ; pathology ; therapy ; Testicular Neoplasms ; blood ; diagnosis ; pathology ; therapy ; Testis ; abnormalities ; alpha-Fetoproteins ; analysis

BACKGROUND: Vascularized bone tissue engineering is a hotspot of current research and early vascularization of tissue-engineered bone also becomes an urgent problem to be solved. Addition of bioactive factors and cell co-culture method both contribute to promoting the early vascularization. OBJECTIVE: To investigate the pro-angiogenic effect of human umbilical vein endothelial cells (HUVECs) transfected with vascular endothelial growth factor 165 (VEGF165) co-cultured with bone marrow mesenchymal stem cells (BMSCs). METHODS: HUVECs were transfected with VEGF165 and then co-cultured with BMSCs which were purified from mouse femoral bone marrow. There were six groups in this experiment: (1) AdVEGF165-HUVECs+BMSCs, (2)AdVEGF165-HUVECs, (3) AdGFP-HUVECs+BMSCs, (4) AdGFP-HUVECs, (5) HUVECs+BMSCs, and (6) HUVECs (blank control). Cell counting kit-8 detection was applied to analyze the proliferative ability of HUVECs. The abilities of HUVECs migration and vascularization were then detected by crystal violet staining and matrigel determination, respectively. RESULTS AND CONCLUSION: Compared with the blank control group, the proliferation of HUVECs was significantly increased in the other groups except for AdGFP-HUVECs group (P < 0.05). Compared with the blank control group, the migration and vascularization abilities of HUVECs were significantly stronger in the group of AdVEGF-HUVECs+BMSCs (P < 0.05). To conclude, the co-culture of HUVECs transfected with VEGF165 and BMSCs could promote early vascularization effectively.

OBJECTIVETo detect hot point mutations of ATP7B gene in Hunan Han patients with Wilson' disease (WD).

METHODSThe genomic DNA of 22 WD patients was extracted and exons 5, 8, 12, 13 were amplified by PCR. Screening for the mutations was done by direct sequencing and analysed by BLAST.

RESULTSFifteen of the 22 patients were found with mutations. Ten heterozygous Arg778Leu (2273G --> T) mutations were found in exon 8, all of them were accompanied with 2250C --> G polymorphism (Leu770Leu). Seven patients were found with 2855G --> A (Arg952Lys) polymorphism (4 heterozygous and 3 homozygous), 3 of them had Arg778Leu mutation in exon 8 and one with heterozygous mutation Gly943Asp (2828G --> A) in exon 12 simultaneously. Only one patient was found with heterozygous Pro992Leu (2975C --> T) mutation in exon 13. No mutations were found in exon 5.

CONCLUSIONArg778Leu is the hot point mutation of ATP7B gene in Hunan Han patients with Wilson' disease while exon 5 is not.

Adenosine Triphosphatases ; genetics ; Adolescent ; Asian Continental Ancestry Group ; genetics ; Cation Transport Proteins ; genetics ; Child ; Copper-transporting ATPases ; DNA ; genetics ; DNA Mutational Analysis ; Exons ; Hepatolenticular Degeneration ; ethnology ; genetics ; Humans ; Mutation

OBJECTIVETo investigate the effects of exposure of paraquat and maneb on the behavior, the morphology and electrical activity of the Substantia nigra and striatum, and to discuss the relationship between this two pesticides and Parkinson's disease.

METHODS37 rats were divided randomly into 3 groups: control group (n = 11), paraquat (10 mg/kg) group (n = 13) and combinative group of paraquat (10 mg/kg) and maneb (30 mg/kg) (n = 13), and were exposed twice a week for 6 weeks by intraperitoneal injection. The behavior of animals in the declined-plane, the vertical-grid and the open-field test were observed. The morphology of substantia nigral neurons were investigated by HE pathology. The spontaneous discharge of striatum neurons were recorded after exposure.

RESULTSCompared to the control group and the pre-exposure group, both the numbers of animals sliding down from the declined-plane and the latency of rats' moving on the vertical-grid significantly increased, and the animals' autonomic movement decreased significantly (P < 0.05, P < 0.001). After the combinative exposure, the neurons of the Substantial nigra pars compacta (SNPc) were progressively impaired, the cell density of the paraquat group [(82.17 ± 12.91) n/mm(2)] and the combined group [(41.15 ± 6.44) n/mm(2)] were lower than that in control group (143.10 ± 20.85 n/mm(2)] (P < 0.01). In the paraquat group (5.97 ± 7.30 Hz) and the combined group [(6.95 ± 9.87) Hz], the average discharge rates of the striatum neurons were increased significantly compared to the control group [(1.78 ± 5.05) Hz] (P < 0.01). The bursting discharge was increased significantly in the combined group (22.3%) compared to the control group (9.8%) and the paraquat group (5.6%) (P < 0.05, P < 0.01).

CONCLUSIONThe co-exposure of paraquat and maneb could induce similar symptoms to Parkinsonism syndrome of rats such as rigidity, moving reduction and etc, and the combined exposure had a certain enhanced effect compared to alone paraquat exposure. The combinative exposure of paraquat and maneb could cause neural loss in SNPc and it is involved with the enhanced electrophysiological activity in striatum. The synergy toxicity of paraquat and maneb in nigrostriatal system is related to Parkinson's disease.

Animals ; Corpus Striatum ; drug effects ; Male ; Maneb ; toxicity ; Paraquat ; toxicity ; Parkinsonian Disorders ; chemically induced ; Pesticides ; toxicity ; Rats ; Rats, Sprague-Dawley ; Substantia Nigra ; drug effects

Adult ; Female ; Humans ; Middle Aged ; Nevus, Blue ; pathology ; Uterine Cervical Neoplasms ; pathology