Objective To study the influence of vascular endothelial growth factor(VEGF) on development of alveolar epithelial cell Ⅱ (AECⅡ) and expression of pulmonary surfactant protein B(SP-B) in premature rats.Methods Wistar rats at 19 days gestation were paunched to get embryo and primary AECⅡculture.The rats were divided into 4 groups ,VEGF-165 group,Dexamethasone group,VEGF and Dexamethason group,control group. AECⅡ and SP-B expression were measured by immunology histochemistry.Results SP-B had positive expression in VEGF group, Dexamethason group, VEGF and Dexamethason group. SP-B had negative expression in control group.Conclusion VEGF-165 can increase SP-B positive expression and secret of AECⅡ.VEGF promotes lung maturity.

Objective Lysine-specific demethylase 1 (LSD1) is a flavin adenine dinucleotide-dependent oxidase, which participates in many biological processes , such as cell proliferation and differentiation and gene activation and repression .The aim of this study was to investigate LSD1 acetylation by histone deacetylase inhib -itor trichostatin A ( TSA) and its effect on TSA-induced apoptosis of ovarian cancer cells . Methods LSD1 shRNA was synthesized and implanted into the pLKO-Tet-On lentiviral vector , which was transfected into HO8910 and SKOV3 ovarian cancer cell lines , and then the transfected cells were screened with 1.5μg/mL puromycin for one week until stable clones were established .The cells were treated for 48 hours with methanol (2 mg/mL, control), TSA (200 nmol/L), TCP (100μmol/L), or TSA+TCP.And in the experiment of RNA interfering the LSD1 expression, the cells were also treated for 48 hours with methanol (2 mg/mL, control), TSA (200 nmol/L), Dox (100 ng/mL), or TSA +Dox.The levels of LSD1 acetylation and its substrate histone H3 lysine 4 dimethylation (H3K4me2) were de-tected by immunoprecipitation (IP) and Western blot.The apoptosis of the cells was determined by Annexin Ⅴ/PI staining and flow cytometry, the transcription levels of the Bax and p21 genes detected by real-time quantitative PCR, and the H3K4me2levels in the promoter regions of Bax and p21 measured by chromatin immunoprecipitation ( ChIP ) .Results In comparison with the methanol control, the TSA group showed significantly increased levels of LSD 1 acetylation in the HO8910(1.00±0.29 vs 5.83±0.46, P<0.01) and SKOV3 cells ( 1.00±0.24 vs 5.07±0.35, P<0.01) as well as that of H3K4me2 ( P<0.01);the total apoptosis rates of HO 8910 and SKOV3 cells were remarkably increased in the TSA, TCP, and TSA+TCP groups (P<0.05), even more significantly in the TSA+TCP than in the TSA and TCP groups ( P<0.05) .The mRNA expressions of Bax and p21 in the HO8910 cells were markedly upregulated in the TSA, Dox, and TSA+Dox groups (P<0.05), even more significantly in the latter than in the former two groups (P<0.05).The TSA group exhibited a higher level of H 3K4me2 than the methanol control in the promoters of Bax(2 .92±0.26 vs 0.68±0.19, P<0.01) and p21 (3.07±0.29 vs 0.93±0.17, P<0.01). Conclusion TSA induces the LSD1 acetylation, while suppression of LSD1 expres-sion and activity may enhance the antitumor activity of TSA .

Objective To study the effects of vascular endothelial growth factor(VEGF)on production of pulmonary surfactants.Method Fetal rat lungs were obtained at 19-day gestation.Primary culture of typeⅡalveolar epithelial cells(AECⅡ)was performed using IgG panning technique.The rats was divided into groups: VEGF,Dexamethasone,VEGF plus Dexamethasone and a control.Total phospholipids,phosphatidylcholine (PC),phosphatidyl glycerol(PG)and sphingornyelin(SM)were determined.Results expressed as mean?SEM. Comparison between groups were made with LSD-t test and one -way ANOVA.Result VEGF,Dexamethasone and VEGF plus Dexamethasone groups showed increased amount of total phospholipids and its compositions on the first day of culture.Conclusions VEGF-165 promotes the production and secretion of pulmonary surfactant. VEGF and Dexamethason may go through different mechanism for enhancement of synthesis of pulmonary surfactant,thereby improve biological function of AECⅡ.

OBJECTIVETo investigate the changes and significance of serum gastrin (GAS), beta-endorphin (beta-EP) and plasma motilin (MTL) in patients with severe burns.

METHODSBlood samples were gotten according to different timepoints from 32 admitted burned patients, and then serum GAS, beta-EP and plasma MTL were determined by radio-immuno assay (RIA).

RESULTSIn patients with severe burns, serum GAS decreased significantly in early period. And at the timepoint of 8 h, it reached the lowest level. But during 9-24 h it elevated for a while, and then it reached a relatively stable level. MTL reached the highest level at the timepoint of 2 h after burning. Then at the shock stage, it was comparatively lower. And at the timepoint of 8 h after burning, it reached the lowest level, then raised persistently after reabsorption, but still lower than the normal level. At the early stage after burning, beta-EP raised, then reached the highest level at 8 h after burning. GAS and MTL decreased and beta-EP increased significantly with the increase of the burned area. However, when the burned area was over 70% of the total body surface area, there was no relationship between them.

CONCLUSIONBlood GAS, MTL and beta-EP have represented regular changes in patients with severe burns at the early stage after burning. And the pain-stimulus and shock are effective factors.

Adolescent ; Adult ; Burns ; blood ; Female ; Gastrins ; blood ; Humans ; Male ; Middle Aged ; Motilin ; blood ; Radioimmunoassay ; Time Factors ; beta-Endorphin ; blood

Objective Protein arginine methyltransferase 5 (PRMT5),a member of the histone arginine methylation transferase family,is involved in a wide range of biological regulation through ei-ther epigenetic or posttranslational methylation modifications. The pur-pose of the present study was to investigate the effects of PRMT5 on cell proliferation of ovarian cancer cell HO8910. Methods Cell lines HO8910 with PRMT5 overexpression were obtained by transi-ent transfection,which were divided into three groups in the experiment: blank control group (wild-type cell line HO8910),negative control group (HO8910 cells were transfected with pCMV-myc plasmid),and experimental group (HO8910 cells were transfected with pCMV-myc-PRMT5 plasmid). Western blot was used to detect the expression of myc protein,and qRT-PCR was used to detect the ex-pression of PRMT5 mRNA. Cell lines HO8910 with inducible stable knockdown of PRMT5 were established by shRNA interference method,which were divided into four groups: pLKO control group (infected by empty vector lentivirus),pLKO+Dox (100ng/mL) group,shPRMT5 group (infected by PRMT5shRNA lentivirus) and shPRMT5+Dox (100 ng/mL) group. Western blot and qRT-PCR were used to detect the expression of PRMT5 protein and mRNA levels. Dox-induced PRMT5 knockdown was detected by increasing Dox concentration,which includes four groups,Dox 0ng/mL group,Dox 1ng/mL group,Dox 10ng/mL group,Dox 100ng/mL group,and each group was treated for 48 hours. Western blot and qRT-PCR were used to detect the PRMT5 protein and mRNA expression. Colony formation assay,EdU assay,and CCK-8 assay were used to test cells proliferation. The experiment was conducted in two large groups each with two subgroups: PRMT5 knockdown group (Dox-group,Dox+ group),PRMT5 overexpression group (pCMV-myc group,pCMV-myc-PRMT5 group). Western blot was used to detect the effects of PRMT5 expression on proliferation-related proteins. The experiment was conducted in two large groups,PRMT5 knockdown group with four subgroups : Dox 0ng/mL group,Dox 1ng/mL group,Dox 10ng/mL group and Dox 100ng/mL group,and PRMT5 overexpression group with two subgroups (pCMV-myc group and pCMV-myc-PRMT5 group). Results Western blot results showed that the expression of myc was detected in the experimental group in which HO8910 cells were transfected with pCMV-myc-PRMT5,and the expression of PRMT5 mRNA was significantly higher in the experimental group than those in the blank control group and the negative control group (P<0.001) . Western blot and qRT-PCR showed that PRMT5 protein (0.32±0.25) and mRNA expression levels in shPRMT5+Dox group were significantly lower than those of shPRMT5 group (0.89±0.18) (P<0.05). Western blot and qRT-PCR confirmed that PRMT5 protein (0.21±0.24) and mRNA expres-sion in Dox 10ng/mL group and Dox 100ng/mL group (0.08±0.15) were significantly downregulated compared to Dox 0ng/mL group (1.11±0.15) (P<0.05). Colony formation experiments,EdU experiments,and CCK-8 experiments confirmed that the proliferative ca-pacity of cells in Dox+group was lower than that of Dox-group in PRMT5 knockdown group(P<0.05); while in PRMT5 overexpression group,the proliferative capacity of pCMV-myc-PRMT5 group was significantly higher than that of the pCMV-myc group (P<0.05). Western blot results showed that the protein expression of Cyclin D1 was significantly lower in Dox 100 ng/mL group (0.17±0.06) than that in Dox 0 ng/mL group (1.18±0.18) (P<0.05) and the expression of P21 was significantly increased in PRMT5 knockdown group (P<0.05). In the PRMT5 overexpression group,the protein expression of Cyclin D1 in pCMV-myc-PRMT5 group (3.48± 0.22) was higher than that in pCMV-myc group (0.88±0.15) (P<0.05),while the protein expression of P21 (0.08±0.17) were significantly lower than that of pCMV-myc group (4.12±0.10) (P<0.05). Conclusion PRMT5 plays an important role in the proliferation of ovarian cancer cells. Down-regulation of PRMT5 can inhibit cell proliferation and up-regulation of PRMT5 can pro-mote cell proliferation.

OBJECTIVETo investigate the characteristics of the phenylalanine hydroxylase (PAH) gene mutations in patients with phenylketonuria (PKU) in Henan province, China, in order for providing basic information for clinical genetic counseling and prenatal diagnosis.

METHODSAll the exons and partial flanking introns of the PAH gene were detected by polymerase chain reaction (PCR) and bi-directional sequencing in 34 patients with PKU from Henan province.

RESULTSA total of 23 different disease-causing mutations were identified which corresponded to 92.65% (63/68) of the PAH alleles, including 12 missense mutations, 4 nonsense mutations, 4 splicing junction mutations, and 3 deletion mutations. Among them, A156P and P69_S70delinsP(delCTT) were novel mutations; IVS2+ 5G to C, G332E, IVS10-14C to G and L367 to Wfs were reported in Chinese population for the first time according to the PAH database (www.pahdb.mcgill.ca). Among all the 13 exons, exon 7 harbored the most type of mutations, exon 11 and exon 5 the second. The most common mutations included R243Q (17.65%, 12/68), V399V (11.76%, 8/68), IVS4-1G to A (8.82%, 6/68), R400T(7.35%, 5/68), Y166X(5.88%,4/68) and G247R(5.88%, 4/68). In addition, 9 other gene variations were found in this study.

CONCLUSIONThe mutation spectrum and frequency of the PAH gene of patients with phenylketonuria in Henan province were slightly different from those from other parts of China.

Asian Continental Ancestry Group ; genetics ; Base Sequence ; Child ; Child, Preschool ; China ; DNA Mutational Analysis ; Female ; Genetic Counseling ; Humans ; Infant ; Male ; Molecular Sequence Data ; Mutation ; genetics ; Phenylalanine Hydroxylase ; genetics ; Phenylketonurias ; diagnosis ; genetics ; Prenatal Diagnosis

OBJECTIVETo evaluate the clinical effect of titanium mesh in anterior cervical subtotal subcorpectomy with locking plate for treatment of cervical spondylotic myelopathy.

METHODSThirty-eight patients with cervical spondylotic myelopathy were treated with anterior cervical subtotal corpectomy using titanium mesh and locking plate. The JOA score of the patients were assessed before and after the operation, and the pre- and postoperative lateral cervical radiographs were taken to observe the instability of the titanium mesh, dynamic plates and changes of the cervical curvature.

RESULTSThe patients were followed up for 12-18 months. Radiographic cervical fusion was achieved in 12-16 months (36 cases) or 18 months (2 cases) postoperatively. The degree of Jordosis was improved and the height of the anterior spinal column and physical curvature were effectively maintained after the operation. The titanium mesh and locking plate showed no signs of loosening and the JOA scores was significantly improved after the operation (P<0.05).

CONCLUSIONTitanium mesh in anterior cervical subtotal corpectomy with locking plate allows effective treatment of cervical spondylotic myelopathy, but the indications of this procedure must be carefully evaluated. The long-term effect of this approach still needs verification by further follow-up data.

Aged ; Bone Plates ; Cervical Vertebrae ; surgery ; Decompression, Surgical ; Female ; Humans ; Male ; Middle Aged ; Orthopedic Procedures ; methods ; Spinal Cord Compression ; etiology ; surgery ; Spinal Fusion ; methods ; Spondylosis ; complications ; surgery ; Surgical Mesh ; Titanium

Acetylcysteine ; therapeutic use ; Animals ; Liver Cirrhosis, Experimental ; drug therapy ; Magnesium Compounds ; therapeutic use ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo compare the effect of high frequency oscillatory ventilation (HFOV) and conventional mandatory ventilation (CMV) on the myocardial function of rabbits with inhalation injury.

METHODSSteam inhalation injury model was reproduced in 16 New Zealand albino rabbits. They were randomly divided into CMV group (n = 8) and HFOV group (n = 8) by drawing lots, and they received ventilation in metered volume and HFOV treatment respectively. Heart blood was drawn from rabbits before they were sacrificed 4 hours after treatment to determine the plasma activity of lactate dehydrogenase 1 (LDH1) and creatine phosphorylated kinase (CPK-MB). Myocardial tissue from left ventricle was harvested and homogenized to determine the concentration of TNF-α and IL-8, the activity of caspase-1, and the activity of myosin-light-chain kinase (MLCK) and the ATPase of myosin light chain (MLC-ATPase) by enzyme-linked immunosorbent assay, spectrophotometry, and the nuclide liquid scintillation technique respectively. Part of the myocardial tissue sample was examined pathologically. Data were processed with analysis of variance.

RESULTS(1) The activities of LDH1 and CPK-MB in plasma were obviously higher in CMV group than in HFOV group [(643 ± 108), (342 ± 48) U vs. (233 ± 92), (186 ± 36) U, with F value respectively 10.326 and 9.846, P values all below 0.01]. (2) The contents of TNF-α, IL-8 and the activity of caspase-1 in myocardial tissue homogenate were obviously higher in CMV group than in HFOV group [(181 ± 35), (89 ± 19) pg/g, and (0.56 ± 0.27) g/g protein vs. (94 ± 21), (43 ± 11) pg/g, and (0.24 ± 0.12) g/g protein, with F value respectively 8.239, 7.826, 5.716, P values all below 0.01]. (3) The activities of MLC-ATPase and MLCK were lower in CMV group than in HFOV group [(0.24 ± 0.12) µmol×mg(-1)×min(-1), (3.3 ± 1.1) mmol×mg(-1)×min(-1) vs. (0.48 ± 0.16) µmol×mg(-1)×min(-1), (7.7 ± 1.7) mmol×mg(-1)×min(-1), with F value respectively 4.125, 4.766, P values all below 0.01]. (4) No obvious necrosis, degeneration or inflammatory cell infiltration was observed in myocardial tissue of rabbits in 2 groups under light microscope; but the myocardial fiber was slightly swollen, and it was less marked in the HFOV group.

CONCLUSIONSThe influence of HFOV on myocardial myosin phosphorylation system of rabbits with inhalation injury is less than that of CMV.

Animals ; Burns, Inhalation ; metabolism ; physiopathology ; therapy ; High-Frequency Ventilation ; Myocardium ; metabolism ; Myosin-Light-Chain Kinase ; metabolism ; Rabbits ; Respiration, Artificial

OBJECTIVETo investigate the effects of high frequency oscillatory ventilation (HFOV) and its combination with administration of pulmonary surfactant (PS) on inflammatory response of lung tissue in rabbits with inhalation injury.

METHODSSevere steam inhalation injury models were reproduced in 24 New Zealand albino rabbits. They were divided into control group (n = 8), HFOV group (n = 8), and HFOV + PS group (n = 8) according to the random number table, and they received ventilation in metered volume, HFOV, and HFOV + PS treatment respectively. Lung tissue samples of rabbits were collected at 3.5 h after treatment for pathological inspection and pulmonary injury score, assay of the activity of myeloperoxidase (MPO) and cysteinyl aspartate-specific protease 1 (caspase-1), and the determination of the contents of TNF-alpha, IL-18, IL-10, IL-13 and their mRNA expression.

RESULTSPathological change in different degree of rabbit lung tissue in each group were observed, and they were most obvious in the control group, and least in the HFOV + PS group. The lung tissue injury scores of control group, HFOV group, and HFOV + PS group was 3.71 +/- 0.43, 2.87 +/- 0.26, and 2.08 +/- 0.28 respectively. The difference between either two of them were statistically significant (P < 0.01). The MPO content and caspase-1 activity of rabbits in HFOV and HFOV + PS groups were obviously lower than those in control group (P < 0.01), and MPO content and caspase-1 activity of rabbits in HFOV + PS group were obviously lower than those in HFOV group (P < 0.05). In HFOV group and HFOV + PS group, the contents of TNF-alpha, IL-18 and their mRNA expression in lung tissue homogenates were obviously lower than those in control group (P < 0.01); while the contents of IL-10, IL-13 and their mRNA expression were obviously higher than those in control group (P < 0.01). Changes in these contents and expression in HFOV + PS group were more obvious than those in HFOV group (P < 0.05).

CONCLUSIONSHFOV can alleviate inflammatory response in rabbit lung tissue and pulmonary injury induced by inhalation injury, and the effect is more obvious when combined with PS.

Animals ; Burns, Inhalation ; complications ; therapy ; Disease Models, Animal ; High-Frequency Ventilation ; Inflammation ; Lung Injury ; etiology ; therapy ; Pulmonary Surfactants ; therapeutic use ; Rabbits