Objective: To clone human angio poietin- related protein-1 angioarrestin genes and construct their recombinant eukaryotic expression vector. Methods: Full length sequence of angioarrestin gene and its C-FD domain were amplified from human liver cDNA library by PCR and were subsequently inserted into the eukaryotic expression vector pcDNA3. 1/ His-Myc (-)B. Then angioarrestin and FD recombinant plasmids were stablely transfected in NCI-H460 cell line. The positive clones were identified by RT-PCR and Western blot. Results: Full length fragment of angioarrestin gene (1 473 bp) and C-FD domain (560 bp) were successfully amplified from human liver cDNA library by PCR. The pcDNA3. 1-ARP and pcDNA3. 1-FD recombinant plasmids were also constructed successfully as identified by PCR and enzyme digestion. RT-PCR and Western blot showed the expression of target mRNA and protein in NCI-1-1460 cells. Conclusion: The eukaryotic expression vectors of angioarrestin and C-FD gene have been successfully constructed and expressed in NCI-H460 cells, which pave a way for further study on the anti-angiogenesis function of angioarrestin in cancer.

Objective To observe the expression of osteoprotegerin (OPG), receptor activator of nuclear factor κβ ligand(RANKL) and receptor activator of nuclear factor κβ (RANK) in bone tissue of rats with chronic fluorosis and to explore the relation between OPG/RANKL/RANK system and bone damage in chronic fluoride poisoning rat and the antagonism effects of Danlan Xianpeng capsule. Methods SD rats were randomly divided into six groups according to body weight (equal male and female in each group): fluorosis group, high dose drug group, medium dose drug group, low dose drug group, control group, borax group(positive control), 12 rats in each group. The control group drank tap water and the remaining 5 experimental groups consumed 50 mg/L fluoride water, and high, medium and low doses drug group took Danlan Xianpeng capsule at doses of 0.8,0.4,0.2 g/kg,borax group took borax at dose of 0.8 g/kg. OPG, RANKL, RANK protein in rat tibial metaphysis was detected by immunohistochemistry at the 6 month. Results Compared with the control group(173.79 ± 5.23, 174.17 ± 5.01,155.63 ± 7.11), the expressions of OPG, RANKL were increased and the expression of RANK was decreased in fluorosis group(156.83 ± 5.80, 157.74 ± 6.70, 173.92 ± 4.37), the difference was statistically significant (all P ＜ 0.05). Compared with the fluorosis group, the expression of OPG and RANKL were decreased and the expression of RANK was increased in high-dose drug group, middle-dose drug group(169.67±5.07, 168.08 ± 5.05,162.12 ± 4.24, 170.78 ± 5.01, 168.41 ± 7.19, 166.69 ± 5.78, all P ＜ 0.05). Compared with the borax group (167.27 ± 4.08, 167.85 ± 5.01, 166.14 ± 3.95), the expression of OPG and RANKL was increased in the low-dose drug group (163.40 ± 4.11, 159.49 ± 5.78), the expression of RANK was increased in the high-dose drug group (162.12 ± 4.24) and decreased in the low-dose drug group(171.54 ± 8.06), the difference was statistically significant (all P ＜ 0.05). Conclusions Chronic fluoride poisoning can cause increased bone turnover and enhance the activity of osteoelastic absorption by increasing RANKL. Danlan Xianpeng capsule can affect bone remodeling through the OPG/RANKL/RANK system, and antagonises bone damage caused by fluoride.

Objective:To investigate the efficacy and safety of less invasive surfactant administration(LISA) of neonatal respiratory distress syndrome(NRDS).Methods:From July 2017 to December 2018, 50 premature infants with birth weight ≤1 500 g and/or gestational age≤32 weeks diagnosed as NRDS at the Fifth Medical Center of PLA General Hospital were randomly divided into LISA group( n=25)and INSURE group( n=25). The patients in LISA group was inserted fine duct into the trachea through direct laryngoscope under nasal continuous positive airway pressure (nCPAP) and pulmonary surfactant was injected.The INSURE group adopts endotracheal intubation-pulmonary surfactant-nCPAP was performed after unplugging.The changes of vital signs, blood gas indexes, adverse reactions and the incidence of complications were compared between two groups at different time points. Results:There was no significant difference in respiration, heart rate, systolic blood pressure, diastolic blood pressure and PaO 2, PaCO 2, BE, SpO 2 between two groups at different administration time points.Although the pH value of LISA group was lower than that of INSURE group, it was within the normal range.There was no significant difference in bronchopulmonary dysplasia, intraventricular hemorrhage, periventricular leucomalacia and other complications between two groups, and there was no death, air leakage, retinopathy of prematurity and pulmonary hemorrhage in both two groups.In addition, there was no significant difference in hospitalization days, total medical expenses, oxygen use time between the two groups(all P>0.05). Conclusion:Compared with INSURE technology, LISA technology has its feasibility for premature infants with NRDS, but the effectiveness and safety in the practical application need to be further confirmed.

Objective To observe the effect of ultrashort wave diathermy on collagen type I expression in early stage of hormon-induced avascular necrosis of the femoral head (ANFH) in rabbit.Methods A total of 40 New Zealand white rabbits were randomly divided into two groups:a control group (n=10) and a treatment group (n =30).All the animals in the treatment group were injected with horse serum and methylprednisolone to establish the ANFH model,and then divided into 2 subgroups:a model group and a ultrashort wave diathermy group,which were treated accordingly.After 12 weeks of treatment,all the animals were sacrificed and collagen type I expression in the femoral head was observed.Results It was shown that the expression of the collagen type I was significantly lower in the model animals than that in the controls as indicated by the stronger immunohistochemistry staining for rabbit collagen type,while that of the ultrashort wave diathermy group was significant higher than the control group ( P

Adult ; Female ; Humans ; Leiomyoma ; complications ; Lung Neoplasms ; complications ; Uterine Neoplasms ; complications

Objective To investigate the clinical pathologic significance of lymphangiogenesis in in colorectal cancer. Methods New lymphatic-specific markers D2-40 was used immunohistochemically to detect the lymphatic vessel density(LVD) in the intratumoural and peritumoral areas, and in normal tissue from 96 cases of colorectal cancer, which were analyzed with clinical pathologic parameters of those colorectal cancer. Results Significandy higher LVD was found in the intratumoural area(14.5±2.4), when compared with normal(5.9±1.1)and peritumoural areas(6.7±1.2) (P<0.01). LVD of the peritumoural area was higher than normal area (P< 0.01). However, peritumoural LVD was associated with both depth of invasion and liver metastasis (r=0.71,0.78 P<0.05), but not associated with tumour size, macroscopic type and lymph-node metastasis (P>0.05). Intratu-moural LVD was not correlated with tumour size, macroscopic type, the depth of invasion,lymph-node metastasis, and liver metastasis(P>0.05). Conclusion Lymphangiogenesis in the peritumoural area may be helpful in evalution of liver metastasis and prognosis.

Objective: To chine and express the recombinant human endothelial monocyte-activating polypeptide-Ⅱ(EMAP-Ⅱ)and identify its anti-tumor biological activities.Methods: EMAP-Ⅱ_(147-312)was expressed by the expression vector pMAL-p2x and E.coli BL-21 and the product was purified.The production of tissue factor(TF)in human umbili- cal vein endothelial cell ECV-304 mediated by the recombinant EMAP-Ⅱwas determined by chemiluminescence sub- strate.The promoting effect of recombinant EMAP-Ⅱon TNF?-induced ECV-304 cell.Apoptosis was determined by flow cytometry.Its inhibitory effect on human pancreaic cancer cell SW1990 proliferation was determined by MTT method. Results:DNA sequencing verified that EMAP-Ⅱwas correctly cloned.The molecular mass of the protein identified by SDS-PAGE was consistent with the theoretic value.The productivity of recombinant EMAP-Ⅱwas 500?g per 1 g bacteria (wet mass).The purified product induced expression of tissue factor(TF)in ECV-304 cells;it also enhanced the sensi- tivity of ECV-304 cells to the apoptotic effect of TNF?([16.6?2.5]% vs[25.6?2.3]%,P

Objective To investigate the effect of hematoxin,lipopolysaccharide(LPS),lipopolysaccharide binding protein(LBP)/membrane CD14(mCD14) in occurrence and development of systemic inflammatory response syndrome(SIRS) in children.Methods Serum LPS,LBP,mean fluorescence intensity(MFI) of mCD14 on monocytes of 30 patients with SIRS were measured,at the same time 21 healthy children had been chosen to serve as control group.Results Compared with control group,the serum LPS,LBP,MFI of mCD14 on monocytes of patients with SIRS were significantly higher(Pa

Objective To create a model with simple expression of mechanical characteristics of the human chest for the development of a manikin. Methods A simplified Gruben-model was proposed based on the anatomical structure and physical characteristics of the materials, and perfect coefficients were computed. The model feasibility was proved by the coefficient of determination and residual analysis.Results The mathematic form of the model provided had three fewer terms than Gruben′s. The coefficient of determination approximated 1, the residue was small, and the perfect coefficients of "a typical human" were determined.Conclusion The hypothesis of the model makes the coefficients physically meaningful, which provides a new method to study the force-displacement relationship of the thorax. Also the simple form makes it easy to create the model and provide some guidance for the design of a manikin′s chest.

OBJECTIVETo investigate mRNA and protein expressions of OPGL and M-CSF mRNA in bones of rats with experimental fluorosis induced by intake of fluoride in the drinking water, and to study the antagonizing effects of Danlan Xianpeng Liaofu capsules on the gene expression.

METHODSTotally 72 SD rats were randomly assorted into 6 groups including the control group, the fluoride group, the high-dosage (0.8 g/kg×d), mid-dosage (0.4 g/kg×d) and low dosage (0.2 g/kg×d) medication groups and the borax group (borax, 0.8 g/kg×d). The distribution of female and male rats in each group was divided up on a fifty-fifty basis. Except the control group, a NaF containing water (NaF 50 mg/L in concentration) was supplied as the drinking water for all the experimental rats in order to establish experimental fluorosis. The thickness and density of trabecula and the thickness of bone cortex were measured by light microscopy. The fluoride content in urine and bone were analyzed by using fluoride ion selective electrode method. Expressions of OPGL and M-CSF mRNA and protein were studied using RT-PCR and immuno-histochemistry, respectively.

RESULTS(1) 10/12 of the experimental fluorosis rats developed dental fluorosis, and 2/12 of dental fluorosis rats occurred in the low-dosage medication group. Fluoride content in urine and bone of the fluorosis rats increased (P<0.05). (2) Compared with that of the control rats, the bone trabecular depth, cortical thickness and trabecular density in experimental fluorosis rats were remarkably reduced. (3) Compared with that of the control group, mRNA expression of both OPGL and M-CSF was increased in the fluoride group rats. The difference was statistically significant (P<0.05). (4) Compared with that of the fluoride group animals, the expression intensity of OPGL mRNA decreased in animals of the control group, the high, mid- and low- dosage medication groups and the borax group. Among them, except the low-dosage group, the difference between all the other groups and the fluoride group was statistically significant, respectively (P<0.05). There was also a decrease of M-CSF mRNA in all the 3 medication groups and the borax group animals in comparing with that of the fluoride group and the difference was also statistically significant (P<0.05), respectively. (5) Compared with that of the control group. There were an increase of OPGL and a decrease of M-CSF protein expression; and in addition, there were a decrease of OPGL and an increase of M-CSF protein expression in all 3 medication groups and the borax group in comparing with that of the fluoride group anima (P < 0.05).

CONCLUSIONSExcessive fluoride induces an accelerated bone turnover and may promote the absorption activity of osteoclasts by increasing the expression of OPGL and M-CSF. Danlan Xianpeng Liaofu capsule may be capable of regulating bone remodeling through a down-regulation on OPGL and M-CSF expression.

Animals ; Bone Density ; drug effects ; Bone Remodeling ; drug effects ; Bone and Bones ; metabolism ; pathology ; Borates ; pharmacology ; Capsules ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; Female ; Fluorides ; metabolism ; urine ; Fluorosis, Dental ; metabolism ; pathology ; Macrophage Colony-Stimulating Factor ; genetics ; metabolism ; Male ; RANK Ligand ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Sodium Fluoride ; poisoning