Objective To learn the gene mutation type and prevalence of thalassemia in the Taishan area and to provide basis for the prevention and treatment of local thalassemia .Methods 1 383 patients who visited the hospital for thalassemia genetic test from January to December 2014 were enrolled for this study .α‐andβ‐genes were determined by using PCR and membrane hybridization methods .Results Of the 1 383 patients ,595 were diagnosed with thalassemia (positive rate was 43 .02% ) ,including 386 cases (27 .91% ) of α‐thalassemia ,183 cases(13 .23% ) of β‐thalassemia and 26 cases(1 .88% ) of αβ‐thalassemia .- -SEA/αα(accounted for 64 .25% ) was the major genotype of α‐thalassemia ,the major genotype of β‐thalassemia wasβ41 -42/β(accounted for 28 .42% ) , and - -SEA/αα,β41 -42/βwas the major genotype of αβ‐thalassemia (accounted for 19 .23% ) .Conclusion Taishan is located in a high risk area of thalassemia ,it is of great significance to perform genetic diagnosis of thalassemia among reproductive population so as to reduce the birth rate of children with thalassemia in the area .

Objective Mitochondrial dysfunction, cell energy metabolism, and oxidative stress play important roles in sepsis-induced acute brain injury.This study was to investigate the effects of Xingnaojing Injection (XNJ) on brain mitochondrial oxidative stress in rats with lipopolysaccharide (LPS)-induced sepsis.Methods Totally, 252 male SD rats were randomly divided into a normal control group, 3 LPS-induced sepsis model groups (LPS 6, 24, and 48 h), and 3 XNJ treatment groups (XNJ 6, 24, and 48 h), with 36 in each group.After treatment, the mitochondrial membrane potential (MMP) was monitored by flow cytometry and the levels of manganese superoxide dismutase (Mn-SOD), malondialdehyde (MDA), nitric oxide (NO), and nitric oxide synthase (NOS) were determined by chromatometry.Results The MMP was significantly increased in the XNJ 6 h group as compared with the LPS 6 h group (0.80±0.11 vs 0.54±0.19, P<0.05).In the LPS and XNJ groups, the levels of MDA and NOS reached the peak at 6 hours and then dropped gradually, while those of NO and Mn-SOD rose to the peak at 24 hours followed by a gradual fall.Statistically significant differences were observed in the levels of MDA, NOS and NO between the LPS 6h and XNJ 6 h groups (P<0.05), as well as in those of NOS, NO and Mn-SOD between the LPS 24 h and XNJ 24 h groups (P<0.05).Conclusion Xingnaojing Injection can elevate the level of the brain mitochondrial membrane potential, improve anti-oxidation indexes in the mitochondria, and protect brain mitochondria in sepsis rats.

Uncoupling protein 2 (UCP2) is a proton transporter which presents in the mitochondrial inner membrane.Recently studies found that UCP2 plays important roles in regulation of reactive oxygen species production,maintenance of mitochondrial function,as well as inflammation and blood glucose control.These features have important relevance with the pathophysiologic mechanism of sepsis.

Objective:To explore whether thioredoin-2 (Trx-2) is involved in the development of cataract and to study the effect of Trx-2 on hydrogen peroxide (H2O2)-induced injury in human lens epithelial cells.Methods:A total of 10 volunteers (removing the lens due totraumatism) and 30 patients received phacoemulsification (age more than 60 years) were selected.The expression of Trx-2 protein in lens epithelial cells from cataract patients and volunteers were detected by the immunohistochemical streptavidin-peroxidase (SP) method.SRA01/04 cells were cultured and were divided into six groups according to different treatment:a control group,H2O2-treated groups at 20,50 or 100 μmol/L,a negative control group (transfected with pCMV6 plasmid plus 100 μmol/L H2O2),and a Trx-2 overexpression group (transfected with pCMV6-Trx-2 plasmid plus 100 μmol/L H2O2).Methyl thiazolyltetrazolium (MTT) assay and flow cytometry was performed to measure the cell viability and apoptosis for SRA01/04 cells,respectively.The activities of superoxide dismutase (SOD) and catalase (CAT),the content of glutathione (GSH) and malondialdehyde (MDA) in human lens epithelial cells were measured via chemical chromatometry.Western blot was used to measure the protein levels of Trx-2,B-cell lymphoma 2 protein (Bcl-2),Bcl-2 associated X protein (Bax) and caspase-3.Results:Compared with the volunteers,the expression of Trx-2 was significantly decreased in lens epithelial cells in patients with cataract (P＜0.05).Compared with the control group,the expression of Trx-2 protein in the 20,50 or 100 μmol/L H2O2 groups was decreased (all P＜0.05).Compared with the control group,the cell survival rates were decreased in the 100 μmol/L H2O2 group and the negative control group (both P＜0.05),along with enhanced apoptotic rates,inhibited cellular SOD activities and CAT activities,reduced GSH contents,augmented MDA contents,down-regulated Trx-2 and Bcl-2 expression and up-regulated Bax and caspase-3 expression (all P＜0.05).Compared with the negative control group,the cell survival rate was increased in the Trx-2 overexpression group (P＜0.05),along with suppressed apoptosis,increased SOD activities and CAT activities,elevated GSH contents,decreased MDA content,up-regulated Trx-2 and Bcl-2 expression and down-regulated Bax and caspase-3 expression (P＜0.05).Conclusion:Trx-2 might be involved in the apoptosis of lens epithelial cells in patients with cataract.The overexpression of Trx-2 obviously attenuated H2O2-induced injury of human lens epithelial cells,which might be associated with the inhibition of H2O2-mediated oxidative stress.

OBJECTIVETo compare therapeutic effects between surgical and conservative treatment for postoperative lumbar discitis.

METHODSFrom January 2004 to January 2009, 41 patients (17 males and 24 females ranging the age from 37 to 68 years with an average of 53.6 years) with postoperative lumbar discitis were retrospectively studied and divided into two groups. There were 19 patients in operation group, 22 patients in conservative group. Clinical data and features,image data, laboratory examinations, antibiotics utilization, hospital stays and sequelae were recorded and analyzed. Visual analogue scales system (VAS) and Oswestry disability index (ODI) were applied to evaluate therapeutic effects.

RESULTSAll patients were followed up over 2 years. Imaging revealed good bone fusion and no occurrence of discitis. VAS score and ODI at 1 month, 1 year and 2 years were significantly improved after treatment (P < 0.01). While VAS and ODI in operation group at 1 month were improved more than that of conservative group (P < 0.01), and there was no significant difference between two groups at 1 year and 2 years (P > 0.01).

CONCLUSIONSurgical and conservative treatment for postoperative lumbar discitis is effective. Surgical treatment is superior to conservative treatment in a short time, while conservative treatment can achieve long-term satisfactory curative effects.

Adult ; Aged ; Anti-Bacterial Agents ; pharmacology ; therapeutic use ; Discitis ; drug therapy ; surgery ; Female ; Humans ; Lumbar Vertebrae ; drug effects ; surgery ; Male ; Middle Aged ; Postoperative Complications ; drug therapy ; surgery ; Retrospective Studies ; Treatment Outcome

Objective To construct a recombinant expression vector for expression of the function-al domains of dengue virus serotype 1 ( DENV1 ) envelope ( E ) protein in native soluble form. Methods The genes encoding the functional domains of DENV1-E protein (1-394 aa) were amplified with PCR and then cloned into the Psectag2B-Fc eukaryotic expression vector.The 293T cells were transfected with the recombinant vector by cationic lipid-based delivery.The cell clones expressing the fusion DENV1-E-Fc protein were screened out with 2 mg/ml of Zeocin.Immunofluorescence assay ( IFA) was performed to analyze the antigenicity and integrity of the fusion protein.The fusion proteins were purified from cell lysate with Protein-G and further identified by Western blot assay.Results The soluble form of fusion protein with a molecular weight of about 90×103 was obtained at a yield of about 25 μg per 1×107 cells.The results of IFA indicated that the fusion protein kept its integrity with right conformational epitopes.The fusion protein was successfully expressed with the advantage of good specificity as indicated by IFA and Western blot assay. Conclusion The recombinant fusion protein in soluble form was successfully expressed in eukaryotic ex-pression system, which paved the way for further investigation on the function of DENV1 E protein and its protective epitopes.

Objective To investigate whether single nucleotide polymorphisms(SNPs)in the ABCC4(rs7320375,rs7329490,rs7986087),FCGR2A(rs1801274)and BLK(rs2254546)region could be susceptibility locus for Kawasaki disease(KD)in children from southern China.Methods This study was performed as a case-control study.The samples,92 individuals with KD and 194 healthy controls from southern China,were collected at Guangzhou Women and Childrens′Medical Center from October,2013 to November,2015,and the SNPs were genotyped by using the Sequenom MassArray system.The genotype distribution and allele frequency of the SNPs were analyzed using chi-square test and Fisher′s exact test.Results The genotype distribution of FCGR2A(rs1801274)in patients with KD were as follows: GG 4.3％(4/92),AG 33.7％(31/92),AA 62％(57/92),correspondingly in healthy controls were GG 48.5％(94/194),AG 41.2％(80/194),AA 10.3％(20/194)respectively,and a significant difference was found between KD patients and controls(x2=98.17,P=0.000).A allele frequency of FCGR2A(rs1801274)in KD patients(78.8％,145/184)was higher than that in controls(30.9％,x2=0.120,P=0.000).The genotype distribution of BLK(rs2254546)in patients with KD were as follows: GG 67.4％(62/92),AG 28.3％(26/92),AA 4.3％(4/92),correspondingly in healthy controls were GG 52.1％(101/194),AG 43.8％(85/194),AA 4.1％(8/194)respectively,significant differences were found between KD patients and controls(x2=6.47,P=0.039).G allele frequency of BLK(rs2254546)in KD patients(81.5％,150/184)was higher than that in controls(74.0％,287/388,x2=1.553,P=0.047).Conclusions For the children in southern China,FCGR2A SNPs(rs1801274)may be associated with the susceptibility to KD,and the A allele may increase the risk of KD.BLK SNPs(rs2254546)is also found to be associated with the susceptibility to KD,and the G allele may increase the risk of KD.

Objective:To investigate the expression status of Sonic Hedgehog signaling genes and molecules in human hepatocellular carcinomas(HCC),and to explore the relationship between these genes and clinical prognosis.Methods:HCC tissue and adjacent normal tissue from 29 HCC patients were assayed for the expression of hedgehog signaling genes by reverse transcription-polymerase chain reaction chain reaction(RT-PCR) techniques and for the expression of hedgehog signaling molecules by immunohistochemistry.The expressions of Shh,Ptch,Smoh,Gli-1 mRNA were assayed as well as Shh,Ptch proteins in 29 cases of HCC and in 29 liver tissues adjacent to the tumor.Results:Expression of Shh mRNA was detectable in about 51% of HCCs examined.Consistent with this,hedgehog target genes Ptch,Smoh and Gli-1 mRNA were expressed in over 68%,48% and 62% of the tumors,respectively,and the expressions of Shh and Ptch proteins in HCC tumor tissues correlated with those of Shh and Ptch mRNA in tumor tissues(P=0.041 and P=0.035).This suggested that the hedgehog pathway was frequently activated in HCCs.The simultaneous expression of Gli-1 in HCC and liver tissues adjacent to the tumor had significantly relationship with poor prognosis.Conclusion:Hedgehog signaling activation is an important event for development of human HCCs.It also suggests that markers for hedgehog signaling activation may be useful for the determination of prognosis.

Objective To constract and evaluate the cationic anticancer peptide Temporin-1CEa liposomes and evaluate anti-breast cancer activity in vitro.Methods The polyethylene glycol (PEG)-modified liposomes containing Temporin-1CEa, one recently discovered cationic anticancer peptide ( CAP) , were constructed by using reverse-phase evaporation method.The encapsulation efficiency, particle size and Zeta potential of the Temporin-1CEa-containing liposomes (Temporin-1CEa-LIP) were characterized.In addition, that had the furhter evaluated of the stability and specific toxicity against human breast cancer MCF-7 cells in vitro.Results The data suggested that the PEG-modified liposomes served a promising drug delivery system for CAPs, those indicated by the encapsulation efficiency was (55.57 ±1.56)%, the particle size was (105.3 ±1.37) nm and the Zeta potential was ( -16.17 ±0.964) mV.Moreover, the in vitro test also indicated that Temporin-1CEa-LIP exerted good stability in serum, and it could be efficiently uptaken by MCF-7 cells.Most importantly, after 24h exposure, Temporin-1CEa-LIP showed toxicity against MCF-7 cells, as potent as Temporin-1CEa. Conclusion The results demonstrates that the PEG-modified liposome is a good drug-delivery system and Temporin-1CEa-LIP could serve as potential anti-tumor candidate for cancer therapy.

OBJECTIVETo develop and evaluate the short version of patient reported outcomes (PROs) questionnaire for gastric stuffiness (Wei Pi) patients with modern test theory and technologies, hoping to provide testing tools for related clinical practice and scientific researches with higher quality and less administrative and response burdens.

METHODSUsing descriptive study design, clinical data were collected with sociological questionnaire and previous developed full items version of PROs instrument for gastric stuffiness (Wei Pi) patients via field and online surveys between Sep 2011 and Mar 2012. The statistical analysis group identified the termination parameters firstly, and then selected items with discrimination, fitting residual, item information curve (IIC) , item characteristic curve (ICC), and the rank of computerized adaptive testing (CAT) select proportion, etc. After assumption evaluation of item response theory (IRT), IIC, ICC, difficulty coefficient distribution, items-response relation and thresholds, etc. were used for psychometric evaluation of instrument.

RESULTSA total of 331 patients [Ages: 31.99 +/- 10.29 yrs; Male: 186 (56.3%)] were enrolled in statistical analysis. The test termination criterion was Max SE = 0.2 or Max items number =16. After items selection, a 15-item short version of instrument, which contains symptoms facet (8 items) and impact facet (7 items) was generated. With good unidimensionality, local independence, and monotonicity, the IC and ICC in IRT analysis showed good working capability of the questionnaire. The difficulty coefficient distribution and items-response relation were also rational, as well as response thresholds.

CONCLUSIONSThe short version of PROs instrument for adult gastric stuffiness (Wei Pi) patients was successfully developed and assessed. The instrument with good methodological and reporting quality could be used in clinical and scientific evaluating their symptoms and impact.

Adult ; Computer Simulation ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Psychometrics ; Stomach Diseases ; diagnosis ; therapy ; Surveys and Questionnaires ; Young Adult