Objective To investigate the interaction of human testis sperm-associated antigen 4 like(SPAG4L) protein containing Sad,UNC-84(SUN) domain with nuclear envelop spectrin repeat proteins 3 (Nesprin-3) containing Klarsicht,ANC-1 and Syne homology (KASH) domain.Methods SPAC-4L protein domains were analyzed with the bioinformatics method.After the SPAG4L plasmid was transfected into Ntera-2 cells,the subcellular localization of SPAG4L was observed.The interaction of SPAG4L with Nesprin-3 was determined by immunofluorescence,co-immunoprecipitation(co-IP) and bimolecular fluorescence complementation (BiFC) technology,respectively.Results Bioinformatics analysis results showed that there was a transmembrane structure in SPAG4L protein.Subcellular localization results demonstrated that SPAG4L protein located in the nuclear membrane and cytoplasm.Immunofluorescence,Co-IP,and BiFC results showed that there was an interaction between SPAG4L and Nesprin-3,and that a LINC(linkers of the nucleoskeleton to the cytoskeletan) complex was formed.Conclusion SPAG4L may interact with Nesprin-3 to form a LINC complex,which is important for understanding the function of SPAG4L protein in spermatogenesis.

ATP Binding Cassette Transporter, Sub-Family B ; metabolism ; Apoptosis ; genetics ; Carcinoma, Non-Small-Cell Lung ; genetics ; metabolism ; pathology ; Genes, MDR ; Humans ; Lung Neoplasms ; genetics ; metabolism ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; RNA, Messenger ; biosynthesis ; genetics ; Telomerase ; biosynthesis ; genetics ; Tumor Suppressor Protein p53 ; metabolism ; Vault Ribonucleoprotein Particles ; metabolism

OBJECTIVETo compare mid-term clinical outcomes between amputation and limb salvage in treating severe open tibial fractures with type Gustilo III B, III C.

METHODSFrom July 2007 to June 2010,68 patients with severe open tibial fractures with type Gustilo III B, III C treated by amputation and limb salvage were retrospectively analyzed. In amputation group, there were 26 males and 12 females with an average age of (44.9±16.3) years old; and 21 cases were type Gustilo (III B, 17 cases were Gustilo III C; amputation were performed in accordance with soft tissue injury degree of shank, fracture types and surgical exploration. In limb salvageg group, there were 21 males and 9 females with an average age of (43.5±14.7) years old; and 23 cases were type Gustilo III B, 7 cases were Gustilo III C; the method of internal fixation and and wound healing were performed in accordance with patients's specific condition. Operative time, blood loss, hospital stay and postoperative infection was compared between two groups; time of loading and rate of return to work was compared; VAS scoring was used to evaluate condition of pain; SF-36 health queationaire was used to assess postoperative life quality.

RESULTSTotally 60 patients were followed up (33 cases in amputation group and 27 cases in limb salvage group) with an average time of 49.1 months. Operative time, blood loss, hospital stay and postoperative infection in amputation and limb salvage group respectively was (109.0±25.7) min, (245.0±58.6) min; (168.0±49.0) ml, (311.0±137.0) ml; (13.8±2.7) d, (28.8±13.1) d; 7.9%, 36.7%. At the final following-up, there was no significance meaning between two groups in VAS scoring and rate of return to work, but time of loading in amputation group was shorter than that of in limb salvage group. Physiological function in amputation group was better than limb salvage group, while body pain was worse; and there was no signicance meaning in psychological health between two groups.

CONCLUSIONAmputation and limb salvage both can treat severe open tibial fractures, and mid-term clinical outcomes between two groups has equivalent efficacy.

Adolescent ; Adult ; Aged ; Amputation ; methods ; Case-Control Studies ; Female ; Humans ; Limb Salvage ; methods ; Male ; Middle Aged ; Tibial Fractures ; surgery

Objective To investigate the expressions of nitric oxide synthase (NOS) protein and mRNA in the lung of rats with hepatopulmonary syndrome. Methods Male Sprague-Dawley rats were divided into four groups: sham operation (SO), intrahepatic portal hypertension (IHPH), prehepatic portal hypertension (PHPH) and portasymstimic shunt (PCS). Two weeks after preparation of rat models, the following measurements were performed: arterial blood gas analysis; the concentrations of NO in lungs; in situ hybridization of ecNOS and iNOS mRNA expressions in lung tissue sections with digoxin-labeled ecNOS and iNOS oligonucleotide probes; expressions of ecNOS and iNOS proteins by immunohistochemisty; image and semiquantitative analysis of the expressions of ecNOS, iNOS and their mRNA. Results PaO_ 2 was (73.85?6.51) mmHg in IHPH rats, significantly lower than that in PHPH, PCS and SO rats97.39? 1.33, 95.23?2.22 and (99.05?0.75)mmHg, respectively.The level of lung NO of IHPH was(19.78?5.33)?mol per gram of protein,much higher than that of PHPH, PCS and SO 13.21?3.99,13.89?3.16 and (8.71?1.68)?mol per gram of protein,respectively. In capillary endothelia, positive expressions of ecNOS mRNA and ecNOS protein in IHPH(4.96?0.82,4.11?0.28) were significantly higher than those of PHPH (1.81? 0.39, 1.63?0.18), PCS (1.88?0.53,1.83?0.16)and SO(1.19?0.32,0.98?0.20). Conclusion The expressions of NOS protein and mRNA in the lung of rats with hepatopulmonary syndrome were increased, and the level of lung NO was elevated, which seems to play an important role in the pathogenesis of hepatopulmonary syndrome.

[ ABSTRACT] AIM:To investigate the effects of sinapic acid ( SA) on the proliferation and apoptosis of rat vas-cular smooth muscle cells (VSMCs) induced by high glucose (HG).METHODS:Cultured A7r5 cells were randomly di-vided and treated as indicated.The cell viability was determined by MTT assay.DNA synthesis was measured by BrdU as-say.Cell cycle progression and cell apoptotic rate were determined by flow cytometry analysis.The levels of reactive oxygen species (ROS) were detected by ELISA.The protein levels of cyclin D1, P21, P27, phosphorylated protein kinase C (p-PKC), p-P38 andβ-actin were evaluated by Western blot.RESULTS:Compared with control group, the viability of A7r5 cells was significantly enhanced, the DNA synthesis was increased, the cell cycle progression was promoted, the levels of ROS were elevated, the cell apoptotic rate was reduced, the protein expression of P21 and P27 was decreased, and the pro-tein levels of cyclin D1, p-PKC and p-P38 were increased in HG group (all P<0.05).These effects were reversed by SA (0.1, 1 and 10 μmol/L) treatment in a dose-dependent manner (all P<0.05).Both P38 inhibitor SB203580 and PKC inhibitor chelerythrine significantly inhibit HG-induced PKC/P38 activation and cell viability ( P <0.05).CONCLU-SION:SA inhibits HG-induced VSMCs proliferation and promotes cell apoptosis via reducing PKC/P38 activation.

OBJECTIVETo explore the DNA methylation levels of genome in cFb transdifferentiation induced by SiO2 in rats.

METHODSThe primary macrophages and fibrocytes of SD rats were co-cultured directly and indirectly, which were exposed to SiO2 at the doses of 25, 50 and 100 g/ml. The transdifferentiation of cFb was identified with immunohistochemical assay. The genomic DNA methylation levels of cFb were detected with HPLC.

RESULTSUnder the condition of indirect co-culture, as compared with control group, the genomic DNA methylation levels of cFb exposed to SiO2 at the doses of 25, 50 and 100 g/ml reduced by 19.9%, 26.9% and 30.3%, respectively (P < 0.05); as compared with cFb exposed to 100 g/ml SiO2, the genomic DNA methylation levels of cFb exposed to 5-aza-dC decreased by 22.0% (P < 0.05). Under the condition of ThinCert(TM) direct co-culture, as compared with control group, the genomic DNA methylation levels of cFb exposed to SiO2 at the doses of 25, 50 and 100 g/ml reduced by 22.2%, 30.2% and 36.7%, respectively (P < 0.05); as compared with cFb exposed to 100 g/ml SiO2, the genomic DNA methylation levels of cFb exposed to 5-aza-dC decreased by 20.6% (P < 0.05).

CONCLUSIONUnder the co-culture condition in vitro, SiO2 could reduce the genomic DNA methylation levels of cFb. The ThinCert(TM) direct co-culture can be used to study the silicosis fibrosis.

Animals ; Cell Transdifferentiation ; drug effects ; Cells, Cultured ; Coculture Techniques ; DNA Methylation ; Fibroblasts ; cytology ; drug effects ; Genome ; drug effects ; Lung ; cytology ; drug effects ; Male ; Rats ; Rats, Sprague-Dawley ; Silicon Dioxide ; adverse effects

Adult ; Case-Control Studies ; Female ; Humans ; Male ; Middle Aged ; Orthopedic Procedures ; Popliteal Cyst ; surgery ; Suture Techniques

OBJECTIVETo compare the short-term clinical outcome between unilateral fixation fusion (ULF) and minimally invasive spine transforaminal lumbar interbody fusion (MIS-TLIF) in treating lumbar disc herniation (LDH).

METHODSThe clinical data of 39 patients with LDH were retrospectively analyzed from June 2008 to March 2013. There was 22 males and 17 females, aged from 45 to 75 years old with an average of 56.9 years. Therer were 3 cases in L3,4, 15 cases in L4,5, 21 cases in L5S1. Among them, 21 patients underwent unilateral fixation fusion (ULF group) and 18 underwent minimally invasive spine transforaminal lumbar interbody fusion (MIS-TLIF group). Operation time, blood loss, the times of radiographic exposure and hospital stay were noted and compared between two groups. Radiograph informations were regularily accessed and VAS, ODI scores were recorded at 3 days and 3, 6, 12 months after operation, respectively. According to modified Macnab criteria, the clinical effects were evaluated at final follow-up.

RESULTSAll operations were successful without severe complications. The averaged operative time and the times of radiographic exposure in ULF group [(95 ± 25) min and (4.2 ± 0.4) times] were less than that of MIS-TLIF group [(120 ± 35) min and (10.1 ± 3.9) times] (P < 0.05). But, the mean blood loss and hospital stay in MIS-TLIF group [(75 ± 45) ml and (7.2 ± 2.2)d ]were less than that of ULF group [(165 ± 60) ml and (11.0 ± 3.7) d] (P < 0.01). All patients were followed up from 12 to 45 months with an average of 29.5 months. The VAS and ODI score had significantly improved during the follow-up and no significant differences were found between two groups at the same time point (P > 0.05). The postoperative radiographs showed internal fixation position was good. And all patients obtained bone fusion by CT scan at 1 year after operation. There was no significant differences in modified Macnab criteria between two groups at the latest follow-up (P > 0.05).

CONCLUSIONFavorable short-term clinical effects can be achieved in suitable LDH patients with ULF or MIS-TLIF surgical procedures.

Aged ; Humans ; Intervertebral Disc Displacement ; surgery ; Lumbar Vertebrae ; surgery ; Middle Aged ; Minimally Invasive Surgical Procedures ; methods ; Spinal Fusion ; methods

To study the effect of Tibetan medicine Zuotai on the activity, protein and mRNA expression of CYP1A2 and NAT2, three different doses (1.2, 3.8 and 12 mg x kg(-1)) of Zuotai were administrated orally to rats once a day or once daily for twelve days, separately. Rats were administrated orally caffeine (CF) on the second day after Zuotai administration, and the urine concentration of CF metabolite 5-acetylamino-6-formylamino-3-methyl-uracil (AFMU), 1-methyluric acid (1U), 1-methylxanthine (1X), 1, 7-dimethylxanthine (17U) at 5 h after study drug administration was determined by RP-HPLC. The activity of CYP1A2 and NAT2 was evaluated by the ratio of metabolites (AFMU+1X+1U)/17U and the ratio of AFMU/(AFMU+1X+1U), respectively. The protein and mRNA expression of CYP1A2 and NAT2 were determined by ELISA and RT-PCR method, respectively. After single administration of Zuotai 3.8 mg x kg(-1) and repeated administration of Zuotai 3.8 and 12 mg x kg(-1), the activity of CYP1A2 and NAT2 decreased significantly compared with control group and there was no significant difference between other dose group and control group. The protein expression of CYP1A2 was significant lower than that in control group after repeated administration of Zuotai 12 mg x kg(-1), and the mRNA expression of CYP1A2 decreased significantly compared with that of control group after single administration of Zuotai 3.8 mg x kg(-1) and repeated admistration of Zuotai 12 mg x kg(-1), separately. The protein expression of NAT2 decreased significantly compared with that of control group after single and repeated administration of Zuotai 3.8 mg x kg(-1), respectively, and the mRNA expression of CYP1A2 decreased significantly compared with control group after single administration of Zuotai 3.8 mg x kg(-1). This study found that Tibetan medicine Zuotai had significant effect on the activity, protein and mRNA expression of CYP1A2 and NAT2.
Administration, Oral ; Animals ; Arylamine N-Acetyltransferase ; genetics ; metabolism ; Caffeine ; metabolism ; urine ; Cytochrome P-450 CYP1A2 ; genetics ; metabolism ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; Female ; Male ; Medicine, Tibetan Traditional ; RNA, Messenger ; metabolism ; Rats ; Rats, Sprague-Dawley ; Theophylline ; urine ; Uracil ; analogs & derivatives ; urine ; Uric Acid ; analogs & derivatives ; urine ; Xanthines ; urine

OBJECTIVETo explore the relationship between the -344T/C polymorphism of aldosterone synthase (CYP11B2) gene and essential hypertension in Chinese Mongolian population.

METHODSBy cluster-sampling method, a total of 1575 Mongolian people in Tongliao city of Inner Mongolia were included in this study. And 417 subjects were normotension, 596 subjects were prehypertension and 562 subjects were essential hypertension. A survey was conducted to collect data by personal interview using a standard questionnaire, meanwhile fasting blood samples were drawn. Height, weight, waist circumference, blood pressure, blood-fat indexes and fasting plasma glucose were measured. The variant genotypes of CYP11B2 were identified by PCR assays. The relationship between the -344T/C polymorphism of CYP11B2 gene and essential hypertension were analyzed by multinomial logistic regression model.

RESULTSCrude prevalence of prehypertension among Mongolian people was 37.84% (596/1575) and hypertension was 35.68% (562/1575). The age-standardized prevalence of prehypertension was 38.57% and hypertension was 31.53%. The frequency of the T and C allele was 0.66 (481/728) and 0.34 (247/728) for normotension group, 0.69 (696/1042) and 0.33 (346/1042) for prehypertension group, 0.71 (706/998) and 0.29 (292/998) for hypertension group. The multiple logistic models showed CYP11B2 variant genotypes were associated with prehypertension (TT/CC, OR = 1.33, 95%CI: 0.87 - 2.01; TC/CC, OR = 1.74, 95%CI: 1.13 - 2.67; TC + TT/CC, OR = 1.49, 95%CI: 1.01 - 2.22); CYP11B2 variant genotypes were associated with hypertension (TT/CC, OR = 1.70, 95%CI: 1.07 - 2.70; TC/CC, OR = 1.59, 95%CI: 0.98 - 2.50; TC + TT/CC, OR = 1.66, 95%CI: 1.06 - 2.58).

CONCLUSIONCYP11B2 gene -344T/C polymorphism were associated with essential hypertension in Chinese Mongolian population.

Adult ; Alleles ; Asian Continental Ancestry Group ; genetics ; Blood Pressure ; genetics ; China ; epidemiology ; Cytochrome P-450 CYP11B2 ; genetics ; Female ; Gene Frequency ; Genotype ; Humans ; Hypertension ; epidemiology ; genetics ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; Young Adult